Protein tyrosine phosphatase wedge domain peptide dimers having non-zero linkers for nervous system repair

Abstract

The disclosure relates to peptide dimers of Formula (I): (I). The disclosure further relates to compositions comprising peptides of Formula (I), methods for nervous system repair, and methods of treating neurological conditions.

Description

[0001] 768698: NGT-003PC

[0002] PROTEIN TYROSINE PHOSPHATASE WEDGE DOMAIN PEPTIDE DIMERS HAVING NON-ZERO LINKERS FOR NERVOUS SYSTEM REPAIR

[0003] RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 702,442 filed on October 2, 2024, the contents of which are hereby incorporated by reference in its entirety.

[0005] BACKGROUND

[0006] Spinal cord injury and other conditions associated with central (CNS) or peripheral (PNS) nervous system damage can cause permanent disability or loss of motor, sensory and / or cognitive function. Recovery after CNS injury is limited, leading to substantial current interest in potential strategies to overcome this challenge. A fundamental obstacle facing efforts to improve neuronal function after injury is the inability of the adult CNS to regenerate.

[0007] Two well-known classes of regeneration inhibitors are myelin- associated inhibitors (e.g., MAG, Nogo and OMGP) and inhibitors in scar tissue formed by glia at the injury site (e.g., chondroitin sulfate proteoglycans (CSPGs)). CSPG deposition causing inhibition of axonal and synaptic repair at sites of nervous system damage has been implicated as a key contributor not only to the pathogenesis of traumatic CNS injury, but to the progression of number of neurodegene rati ve and neuroinflammatory diseases as well.

[0008] CSPGs present a barrier to axon regeneration via several inhibitory mechanisms. The inhibitory effects of CSPG are not only reflected in the formation of dystrophic axonal retraction bulbs that fail to regenerate through the lesion, but also in the limited ability for collateral sprouting of spared fibers. Although it has been known for nearly two decades that sulfated proteoglycans are major contributors to the repulsive nature of the glial scar, the precise inhibitory mechanism was poorly understood.

[0009] Protein tyrosine phosphatases (PTPs) play an important role during dephosphorylation, a process that can remove phosphoryl groups from phosphotyrosinecontaining proteins (Jing-Fei Huang, Molecular Biology and Evolution, Volume 20, Issue 5, May 2003, Pages 815-820). Receptor-type protein tyrosine phosphatases (PTPRs) are a subgroup of PTPs that share a transmembrane domain with resulting similarities in function and target specificity (Du Y, Grandis JR. Chin J Cancer. 2015;34(2):61-69).

[0010] The leukocyte common antigen related (LAR) subfamily PTPRs consists of three members: LAR (PTPRF), receptor protein tyrosine phosphatase sigma (PTPRS) and receptor protein tyrosine phosphatase delta (PTPD). PTPRS and PTPF have been identified as receptors for CSPGs, the principal inhibitory constituents of the glial scar and perineuronal net. The sugar side chains of CSPGs can bind to PTPRF and PTPRS 768698: NGT-003PC expressed by cells, such as neural cells, and inhibit neural cell growth, plasticity, regeneration and sprouting failure in the neural cells.

[0011] It was found that PTPRS-deficient neurons exhibit decreased sensitivity to CSPG- mediated inhibition in various cell-based assays and showed increased regeneration following neurological injury, such as following spinal cord injury and optic nerve crush. The results in the PTPRF knockout remained inconclusive, with both increased and decreased regenerative phenotypes being found. Since CSPGs are the primary impediment to regeneration and plasticity in the injured adult nervous system, modulators of the LAR family protein tyrosine phosphatase functions can be used as therapeutic agents promoting neural plasticity, regeneration and ultimately repair of nervous system damage.

[0012] In view of the foregoing, there remains an urgent need for compositions that modulate and attenuate the inhibitory CSPG function. In addition, there remains a need for compositions that can alleviate CSPG-induced cellular and neurologic deficits associated with the function of the LAR family protein tyrosine phosphatases.

[0013] SUMMARY

[0014] The present disclosure provides, inter alia, a peptide dimer of Formula (I):

[0015] (I), or a pharmaceutically acceptable salt thereof, wherein:

[0016] X1ais a first peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of TAT;

[0017] X2ais a second peptidyl domain comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR);

[0018] X1bis a third peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of TAT;

[0019] X2bis a fourth peptidyl domain comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR); 768698: NGT-003PC

[0020] R is -C(i-6)alkylene- or -C(i-6)alkylene-[OCH2CH2]n-O-C(i-6)alkylene-; and n is 1-20.

[0021] In some embodiments, the first peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, the first peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3. In some embodiments, the first peptidyl domain comprises the amino acid sequence of SEQ ID NO: 1 .

[0022] In some embodiments, the second peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, the second peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, and 15. In some embodiments, the second peptidyl domain comprises the amino acid sequence of SEQ ID NO: 14.

[0023] In some embodiments, the third peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, the third peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3. In some embodiments, the third peptidyl domain comprises the amino acid sequence of SEQ ID NO: 1 .

[0024] In some embodiments, the fourth peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, the fourth peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, and 15. In some embodiments, the fourth peptidyl domain comprises the amino acid sequence of SEQ ID NO: 14.

[0025] In some embodiments, the present disclosure provides a peptide dimer of Formula (la): 768698: NGT-003PC

[0026] GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH 2

[0027] I L

[0028] I

[0029] GRKKRRQRRR . DMAEHTERLKANDSLKLSQEYESI-NH2

[0030] (la), or a pharmaceutically acceptable salt thereof.

[0031] In some embodiments, R is -C(i.6)alkylene-. In some embodiments, R is -CH2CH2-. In some embodiments, R is -C(i.2)alkylene-[OCH2CH2]n-O-C(i.2)alkylene-, and n is 1-6. In some embodiments, R is -CH2CH2OCH2CH2OCH2CH2-.

[0032] In some embodiments, the present disclosure provides a peptide dimer selected from the group consisting of:

[0033] H H GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH2

[0034] GRKKRRQRRR . DMAEHTERLKANDSLKLSQEYESI-NH2

[0035] KO and

[0036] 768698: NGT-003PC

[0037] H H hL A

[0038] GRKKRRQRRR' DMAEHTERLKANDSLKLSQEYESI-NH2 or a pharmaceutically acceptable salt thereof.

[0039] The present disclosure further provides a pharmaceutical composition comprising a peptide dimer described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0040] The present disclosure further provides a method of repairing the nervous system and / or treating a neurological condition, disease or disorder selected from the group consisting of neural injury, neurological disease caused by inflammation or autoimmunity, and neurodegenerative disease in a subject in need thereof, the method comprising administering an effective amount of a pharmaceutical composition described herein.

[0041] In some embodiments, the neural injury is selected from the group consisting of acute neural injury, traumatic brain injury (TBI), spinal cord injury, concussion, stroke, including ischemic stroke, hemorrhagic stroke, chronic stroke disease, aneurysm, cerebral hemorrhage, thrombus, and embolism.

[0042] In some embodiments, the neurological condition, disease or disorder is selected from the group consisting of Alzheimer’s Disease, dementias related to Alzheimer’s Disease, Lewy diffuse body diseases, senile dementia, Parkinson’s Disease, amyotrophic lateral sclerosis, multiple sclerosis (MS), optic neuritis, Huntington’s Disease, Tourette’s syndrome, hereditary motor and sensory neuropathy, diabetic neuropathy, progressive supranuclear palsy, Jakob-Creutzfeldt disease, epilepsy, and infectious disease

[0043] The present disclosure further provides the use of a pharmaceutical composition described herein in the manufacture of a medicament for treatment of a neurological condition, disease or disorder selected from the group consisting of neural injury, neurological disease caused by inflammation or autoimmunity, and neurodegenerative disease. 768698: NGT-003PC

[0044] BRIEF DESCRIPTION OF THE DRAWINGS

[0045] The accompanying drawings, which are incorporated herein and constitute part of this specification, illustrate exemplary embodiments of the disclosure, and, together with the description given below, serve to explain the features of the present disclosure.

[0046] FIG. 1 is a set of transmission electron microscopy images illustrating the distinct patterns of self-assembly of Compounds 2 and 3 in water and isotonic saline.

[0047] DETAILED DESCRIPTION

[0048] Embodiments of the present disclosure relate to peptide dimers, compositions comprising the peptide dimers, and methods of using the peptide dimers for repairing the nervous system of a subject or for treating diseases, disorders, and / or conditions associated with activation and signaling of the LAR family of phosphatases.

[0049] Definitions

[0050] Listed below are definitions of various terms used to describe the compounds and compositions disclosed herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0051] Unless otherwise defined, all scientific and technical terms used herein shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Generally, nomenclature utilized in connection with, and techniques of, cell and tissue culture, molecular biology, and protein and oligo- or polynucleotide chemistry and hybridization described herein are those well-known and commonly used in the art.

[0052] As used herein, “one or more of a, b, and c” means a, b, c, ab, ac, be, or abc. The use of “or” herein is the inclusive or.

[0053] As used in the specification and in the claims, the “comprise(s),” “comprising,” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named features, groups, ingredients, or steps and does not exclude the presence of additional features, groups, ingredients, or steps. The term “comprise(s),” “comprising,” “include(s),” “having,” “has,” “can,” or “contain(s),” can include embodiments encompassed by the term "consisting essentially of" or "consisting of."

[0054] As used herein, the term “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20% or ±10%, including ±5%, ±1%, and 768698: NGT-003PC

[0055] ±0.1 % from the specified value, as such variations are appropriate to perform the disclosed methods.

[0056] Chemistry

[0057] As used herein, the term “alkyl” refers to a straight or branched saturated hydrocarbon. For example, an alkyl group can have 1 to 12 carbon atoms (i.e., (Ci- Ci2)alkyl), 1 to 6 carbon atoms (i.e., (Ci-C6)alkyl), 1 to 4 carbon atoms (i.e., (Ci-C4)alkyl), or 1 to 3 carbon atoms (i.e., (Ci-C3)alkyl). Examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1 -propyl (n-Pr, n-propyl, -CH2CH2CH3), isopropyl ( / -Pr, / -propyl, -CH(CH3)2), 1 -butyl (n-bu, n-butyl, -CH2CH2CH2CH3), 2-butyl (s-bu, s-butyl, - CH(CH3)CH2CH3), tert-butyl (t-bu, t-butyl, -CH(CH3)3), 1 -pentyl (n-pentyl, - CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3) CH2CH2CH3), neopentyl (-CH2C(CH3)3), 1 -hexyl (- CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), heptyl (-(CH2)6CH3), octyl (- (CH2)7CH3), 2,2,4-trimethylpentyl (-CH2C(CH3)2CH2CH(CH3)2), nonyl (-(CH2)8CH3), decyl (- (CH2)9CH3), undecyl (-(CH2)I0CH3), and dodecyl (-(CH2)nCH3). In an embodiment, alkyl refers to C(i.6)alkyl. In another embodiment, alkyl refers to C^alkyl. In another embodiment, alkyl refers to C(i-3>alkyl.

[0058] As used herein, the term “alkylene” refers to a bivalent alkyl group. For example, an alkylene group can have 1 to 12 carbon atoms (i.e., (Ci-Ci2)alkylene), 1 to 6 carbon atoms (i.e., (Ci-C6)alkylene), 1 to 2 carbon atoms (i.e., (Ci-C2)alkylene), or 1 carbon atom (i.e., (Ci)alkylene). Examples of alkylene groups include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), n-propylene (-CH2CH2CH2-), n-butylene (-CH2CH2CH2CH2-), etc.

[0059] As used herein, the term “PEG” is used broadly to encompass any polyethylene glycol molecule, without regard to size or to modification at an end of the PEG, unless otherwise specified. In some embodiments, PEG is represented by the general formula (- CH2CH2O-)n, wherein n is an integer representing the number of ethylene glycol repeating units. PEGs can vary in molecular weight depending on the value of n, and may be linear or branched.

[0060] Polypeptides

[0061] As used herein, the term “wild type” refers to the naturally occurring protein sequence, or portion thereof, respectively, as it normally exists in vivo.

[0062] As used herein, the term “recombinant” means that a protein is derived from a prokaryotic or eukaryotic expression system. As used herein, the term “recombinant” refers to a genetic material formed by a genetic recombination process. A “recombinant protein” is made through genetic engineering. A recombinant protein is coded by a DNA sequence created artificially. A recombinant protein is a protein that is coded by a recombinant nucleic acid sequence. A recombinant nucleic acid sequence has a sequence from two or more sources incorporated into a single molecule. As used herein, the term “nucleic acid” refers to 768698: NGT-003PC polynucleotides, such as deoxyribonucleic acid (DNA), and, where appropriate, ribonucleic acid (RNA). The term should also be understood to include, as equivalents, analogs of either RNA or DNA made from nucleotide analogs, and, as applicable to the embodiment being described, single (sense or antisense) and double-stranded polynucleotides.

[0063] As used herein, the term “expression cassette” refers to a part of a vector DNA used for cloning and transformation. In each successful transformation, the expression cassette directs the cell’s machinery to make polypeptide. Some expression cassettes are designed for modular cloning of protein-encoding sequences so that the same cassette can easily be altered to make different proteins. Expression cassettes may also refer to a recombinantly produced nucleic acid molecule that is capable of expressing a genetic sequence in a cell. An expression cassette typically includes a regulatory region such as a promoter, (allowing transcription initiation), and a sequence encoding one or more proteins or RNAs. Optionally, the expression cassette may include transcriptional enhancers, non-coding sequences, splicing signals, transcription termination signals, and polyadenylation signals. The sequences controlling the expression of the gene, i.e. its transcription and the translation of the transcription product, are commonly referred to as regulatory unit. Most parts of the regulatory unit are located upstream of coding sequence of the heterologous gene and are operably linked thereto. The expression cassette may also contain a downstream 3’ untranslated region comprising a polyadenylation site. The regulatory unit of the present disclosure is either directly linked to the gene to be expressed, i.e. transcription unit, or is separated therefrom by intervening DNA such as for example by the 5’-untranslated region of the heterologous gene. Preferably the expression cassette is flanked by one or more suitable restriction sites in order to enable the insertion of the expression cassette into a vector and / or its excision from a vector. Thus, the expression cassette according to the present disclosure can be used for the construction of an expression vector, in particular a mammalian expression vector.

[0064] As used herein, the term “expression vector,” otherwise known as an expression construct, refers to a plasmid or virus designed for protein expression in cells. The vector is used to introduce a specific gene into a target cell and can commandeer the cell’s mechanism for protein synthesis to produce the protein encoded by the gene. The plasmid is engineered to contain regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene carried on the expression vector. The goal of a well- designed expression vector is the production of significant amount of stable messenger RNA, and therefore proteins.

[0065] As used herein, the term “host cell” and the term “host” refer to 1 ) a cell that harbors foreign molecules, viruses, etc.; 2) a cell that has been introduced with DNA or RNA, such as a bacterial cell acting as a host cell for the DNA isolated from a bacteriophage. 768698: NGT-003PC

[0066] As used herein, a “fusion” or “chimeric” protein comprises a first amino acid sequence linked to a second amino acid sequence with which it is not naturally linked in nature. The amino acid sequences which normally exist in separate proteins can be brought together in the fusion polypeptide, or the amino acid sequences which normally exist in the same protein can be placed in a new arrangement in the fusion polypeptide, e.g., fusion of a PTPR wedge domain sequence with transport moiety sequence. A fusion protein may be created, for example, by chemical synthesis, or by creating and translating a polynucleotide in which the peptide regions are encoded in the desired relationship. A chimeric protein can further comprise a second amino acid sequence associated with the first amino acid sequence by a covalent, non-peptide bond or a non-covalent bond.

[0067] As used herein, the term “modified” and the term “mutant” when made in reference to a gene or to a gene product refer, respectively, to a gene or to a gene product which displays modifications in sequence and / or functional properties (i.e., altered characteristics) when compared to the wild-type gene or gene product.

[0068] As used herein, the term “amino acid” includes alanine (Ala or A); arginine (Arg or R); asparagine (Asn or N); aspartic acid (Asp or D); cysteine (Cys or C); glutamine (Gin or Q); glutamic acid (Glu or E); glycine (Gly or G); histidine (His or H); isoleucine (lie or I): leucine (Leu or L); lysine (Lys or K); methionine (Met or M); phenylalanine (Phe or F); proline (Pro or P); serine (Ser or S); threonine (Thr or T); tryptophan (Trp or W); tyrosine (Tyr orY); and valine (Vai orV). Non-traditional amino acids are also within the scope of the disclosure and include norleucine, ornithine, norvaline, homoserine, and other amino acid residue analogues such as those described in Ellman et al. Meth. Enzym. 202:301-336 (1991 ). To generate such non-naturally occurring amino acid residues, the procedures of Noren et al. Science 244:182 (1989) and Ellman et al., supra, can be used. Briefly, these procedures involve chemically activating a suppressor tRNA with a non-naturally occurring amino acid residue followed by in vitro transcription and translation of the RNA. Introduction of the non- traditional amino acid can also be achieved using peptide chemistries known in the art. As used herein, the term “polar amino acid” includes amino acids that have net zero charge but have non-zero partial charges in different portions of their side chains (e.g., M, F, W, S, Y, N, Q, C). These amino acids can participate in hydrophobic interactions and electrostatic interactions. As used herein, the term “charged amino acid” includes amino acids that can have non-zero net charge on their side chains (e.g., R, K, H, E, D). These amino acids can participate in hydrophobic interactions and electrostatic interactions.

[0069] As used herein, the terms “peptide” or “polypeptide” are used interchangeably herein and refer to compounds consisting of from about 2 to about 90 amino acid residues, inclusive, wherein the amino group of one amino acid is linked to the carboxyl group of another amino acid by a peptide bond. A peptide can be, for example, derived or removed 768698: NGT-003PC from a native protein by enzymatic or chemical cleavage, or can be prepared using conventional peptide synthesis techniques (e.g., solid phase synthesis) or molecular biology techniques (see Sambrook et al., MOLECULAR CLONING: LAB. MANUAL (Cold Spring Harbor Press, Cold Spring Harbor, NY, 1989)). A “peptide” can comprise any suitable L- and / or D-amino acid, for example, common a-amino acids (e.g., alanine, glycine, valine), non-a-amino acids (e.g., P-alanine, 4-aminobutyric acid, 6 aminocaproic acid, sarcosine, statine), and unusual amino acids (e.g., citrulline, homocitrulline, homoserine, norleucine, norvaline, ornithine). The amino, carboxyl and / or other functional groups on a peptide can be free (e.g., unmodified) or protected with a suitable protecting group. Suitable protecting groups for amino and carboxyl groups, and means for adding or removing protecting groups are known in the art. See, e.g., Green & Wuts, PROTECTING GROUPS IN ORGANIC SYNTHESIS (John Wiley & Sons, 1991 ). The functional groups of a peptide can also be derivatized (e.g., alkylated) using art-known methods.

[0070] As is clear to the skilled artisan, the peptide sequences disclosed herein are shown proceeding from left to right, with the left end of the sequence being the N-terminus of the peptide and the right end of the sequence being the C-terminus of the peptide. Among sequences disclosed herein are sequences incorporating an “ — OH” moiety or an “ — NH2” moiety at the carboxy terminus (C-terminus) of the sequence. In such cases, and unless otherwise indicated, an “ — OH” or an “ — NH2” moiety at the C-terminus of the sequence indicates a hydroxy group or an amino group, corresponding to the presence of an amido (CONH2) group at the C-terminus, respectively. In each sequence of the present disclosure, a C-terminal “ — OH” moiety may be substituted for a C-terminal “ — NH2” moiety, and vice- versa.

[0071] As used herein, the term “monomer” or “peptide monomer” refers to a peptide molecule that may bind chemically to other molecules such as another peptide molecule to form a polymer.

[0072] As used herein, the term “peptide dimer” refers broadly to a peptide molecule comprising two monomer subunits, which can be identical or different. As such, dimers of the present disclosure include homodimers and heterodimers.

[0073] As used herein, the term “subunit” refers to a separate polypeptide chain that makes a certain protein which is made up of two or more polypeptide chains joined together. In a protein molecule composed of more than one subunit, each subunit can form a stable folded structure by itself. The amino acid sequences of subunits of a protein or polypeptide can be identical, similar, or completely different.

[0074] As used herein, the term “linker” and the term “peptide linker” are interchangeable and refer to short peptide sequences that occur between functional protein domains and link the functional domains together. Linkers designed by researchers are generally classified 768698: NGT-003PC into three categories according to their structures: flexible linkers, rigid linkers, and in vivo cleavable linkers. A flexible linker is often composed of flexible residues like glycine and serine so that the adjacent protein domains are free to move relative to one another. A linker also may play a role in releasing the free functional domain in vivo (as in in vivo cleavable linkers). Linkers may offer many other advantages for the production of fusion proteins, such as improving biological activity, increasing expression yield, and achieving desirable pharmacokinetic profiles. The composition and length of a linker may be determined in accordance with methods well known in the art and may be tested for efficacy. A linker may be from about 3 to about 15 amino acids long. In some embodiments of the present invention, a linker may be about 5 to about 10 amino acids long, however, longer linker may be used in embodiments of the present invention.

[0075] As used herein, the terms “portion,” “fragment,” “variant,” “derivative,” and “analog,” when referring to a polypeptide of the present disclosure, include any polypeptide that retains at least some biological activity referred to herein (e.g., inhibition of an interaction such as binding). Polypeptides as described herein may include portion, fragment, variant, or derivative molecules without limitation, as long as the polypeptide still serves its function. Polypeptides or portions thereof of the present disclosure may include proteolytic fragments, deletion fragments and in particular, or fragments that more easily reach the site of action when delivered to an animal.

[0076] As used herein, the term “protein purification” refers to a series of processes intended to isolate one or a few proteins or polypeptides from a complex mixture, such as cell culture media, cells, tissues or whole organisms, etc. Usually, a protein purification protocol contains one or more chromatographic steps. The basic procedure in chromatography is to flow the solution containing the protein through a column packed with various materials. Different proteins interact differently with the column material and can thus be separated by the time required to pass the column, or the conditions required to elute the protein from the column. Many purification strategies exist. For example, a protein can be attached with an antigen peptide tag by engineering and be purified using an antibody against the antigen peptide tag. Usually, during purification, the protein with an antigen peptide tag can be added on a column loaded with resin that is coated with an antibody or by incubating with a loose resin that is coated with an immobilizing antibody. This particular procedure is known as immunoprecipitation. Immunoprecipitation is quite capable of generating an extremely specific interaction which usually results in binding only the desired protein. The purified tagged proteins can then easily be separated from the other proteins in solution and later eluted back into clean solution.

[0077] In some embodiments, the dimers disclosed herein are substantially isolated. By “substantially isolated” it is meant that the dimer is at least partially or substantially separated 768698: NGT-003PC from the environment in which it was formed or detected. Partial separation can include, for example, a dimer enriched in the compound of the present disclosure. Substantial separation can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the dimer.

[0078] As used herein, the term “inhibitor” refers to a molecule, compound, or agent that reduces or inhibits at least one activity, signaling, or function of leukocyte-common antigen related (LAR) family of phosphatases induced by proteoglycan, reduces in the activity, signaling, and / or function of chondroitin sulfate proteoglycan (CSPG), and / or the interaction between chondroitin sulfate proteoglycan (CSPG) and LAR family of phosphatases. In some embodiments, “inhibitor” also refers to a molecule, compound, or agent that abolish inhibitory effects of CSPGs on neural cells activated with CSPGs. In various embodiments, inhibitors disclosed herein are peptide dimers comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR).

[0079] As used herein “TAT” refers to a cell-penetrating protein that is encoded by the tat gene in human immunodeficiency virus 1 (HIV-1 ). TAT consists of between 86 and 101 amino acids depending on the subtype.

[0080] Homology

[0081] As used herein, the terms “homology” and “identity” are used synonymously throughout and refer to sequence similarity between two peptides. Homology can be determined by comparing a position in each sequence, which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same amino acid, then the molecules are homologous or identical at that position. A degree of homology or identity between sequences is a function of the number of matching or homologous positions shared by the sequences.

[0082] As used herein, the term “analogue” and the term “analog” refer to one of a group of chemical compounds that share structural and / or functional similarities but are different in respect to elemental composition. A structural analog is a compound having a structure similar to that of another one, but differing from it in respect of one or more components, such as one or more atoms, functional groups, or substructures, etc. Functional analogs are compounds that have similar physical, chemical, biochemical, or pharmacological properties. Functional analogs are not necessarily also structural analogs with a similar chemical structure.

[0083] As used herein, the term “sequence identity,” “percent identity,” “percent homology,” or, for example, comprising a “sequence 80% identical to,” refer to the extent that sequences are identical on an amino acid-by-amino acid basis over a window of comparison. Thus, a “percentage of sequence identity” may be calculated by comparing two optimally aligned 768698: NGT-003PC sequences over the window of comparison, determining the number of positions at which the the identical amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Vai, Leu, lie, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gin, Cys and Met) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity.

[0084] Calculations of sequence similarity or sequence identity between sequences (the terms are used interchangeably herein) can be performed as follows. To determine the percent identity of two amino acid sequences, the sequences can be aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In certain embodiments, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues at corresponding amino acid positions are then compared. When a position in the first sequence is occupied by the same amino acid residue as the corresponding position in the second sequence, then the molecules are identical at that position.

[0085] The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0086] Alternately, the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In some embodiments, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch, (1970, J. Mol. Biol. 48: 444-453) algorithm which has been incorporated into the GAP program in the GCG software package, using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1 , 2, 3, 4, 5, or 6. Another exemplary set of parameters includes a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (1989, Cabios, 4: 11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.

[0087] For instance, the peptide sequences described herein can be used as a “query sequence” to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al., (1990, J. Mol. Biol, 215: 403-10). BLAST 768698: NGT-003PC protein searches can be performed with the XBLAST program, score=50, wordlength=3 to obtain amino acid sequences homologous to protein molecules of the present disclosure. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (Nucleic Acids Res. 25:3389-3402, 1997). When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0088] Compositions and Formulations

[0089] As used herein, the term “pharmaceutically acceptable” refers to a compound or drug approved or approvable by a regulatory agency of a federal or a state government, listed or listable in the U.S. Pharmacopeia or in other generally recognized pharmacopeia for use in mammals, including humans.

[0090] As used herein, the term “pharmaceutically acceptable salt” refers to derivatives of the disclosed compounds wherein the parent compound is modified by converting an existing acid or base moiety to its salt form. Lists of suitable salts are found in Remington’s Pharmaceutical Sciences, 17thed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0091] As used herein, the term “composition” or “pharmaceutical composition” refers to a mixture of at least one compound useful within the disclosure with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, and topical administration.

[0092] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the disclosure within or to the subject such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the disclosure, and not injurious to the subject. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean 768698: NGT-003PC oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0093] As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the present disclosure, and that are physiologically acceptable to the subject. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound disclosed herein. Other additional ingredients that may be included in the pharmaceutical compositions are known in the art and described, for example, in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

[0094] As used herein, the term “pharmaceutical formulation” and the term “drug formulation” refer to a mixture or a structure in which different chemical substances, including the active drug, are combined to form a final medicinal product, such as a sterile product, a solution, a powder, an emulsion, a capsule, a tablet, a granule, a topical preparation, a non-conventional product such as semi-solid or sustained-release preparations, liquid, etc. Pharmaceutical formulation is prepared according to a specific procedure, a “formula.” The drug formed varies by the route of administration.

[0095] Medical Intervention

[0096] As used herein, the term “dose” refers to a specified amount of medication taken at one time. A “daily dose” refers to the total dosage amount administered to an individual in a single 24-hour day.

[0097] As used herein, the term “mg / kg” refers to the dose of a substance administered to an individual in milligrams per kilogram of body weight of the individual.

[0098] As used herein, the term “dosage” refers to the administering of a specific amount, number, and frequency of doses over a specified period of time. Dosage implies duration. A “dosage regimen” is a treatment plan for administering a drug over a period of time.

[0099] As used herein, the phrases “parenteral administration” and “administered parenterally” as used herein means modes of administration other than enteral and topical administration or through the digestive tract, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, 768698: NGT-003PC subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intracisternal injection and infusion.

[0100] As used herein, the phrases “systemic administration,” “administered systemically,” “peripheral administration” and “administered peripherally” as used herein mean the administration of a compound, drug or other material other than directly into a target tissue (e.g., the nervous system), such that it enters the animal’s system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.

[0101] As used herein, the term “patient” or “subject” or “animal” or “host” refers to any mammal. The subject may be a human but can also be a mammal in need of veterinary treatment, e.g., domestic animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, sheep, fowl, pigs, horses, and the like) and laboratory animals (e.g., rats, mice, guinea pigs, and the like).

[0102] As used herein, the term “administering” to a patient includes dispensing, delivering or applying an active compound in a pharmaceutical formulation to a subject by any suitable route for delivery of the active compound to the desired location in the subject (e.g., to thereby contact a desired cell such as a desired neuron), including administration into the cerebrospinal fluid or across the blood-brain barrier, delivery by either the parenteral or oral route, intramuscular injection, subcutaneous or intradermal injection, intravenous injection, buccal administration, transdermal delivery and administration by the rectal, colonic, vaginal, intranasal or respiratory tract route. The agents may, for example, be administered to a comatose, anesthetized, or paralyzed subject via an intravenous injection or may be administered intravenously to a pregnant subject to stimulate axonal growth in a fetus. Specific routes of administration may include topical application (such as by eyedrops, creams or erodible formulations to be placed under the eyelid, intraocular injection into the aqueous or the vitreous humor, injection into the external layers of the eye, such as via subconjunctival injection or subtenon injection, parenteral administration or via oral routes.

[0103] The term “treat,” “treated,” “treating,” or “treatment” includes the diminishment or alleviation of at least one symptom associated or caused by the state, disorder or disease being treated. The term “treatment” as used herein also includes: (1 ) inhibiting the disease or condition, i.e., arresting the development or progression of the disease or condition, (2) relieving the disease or condition, i.e., causing the condition to regress, (3) stopping the symptoms of the disease, and / or (4) enhancing the conditions desired.

[0104] As used herein, the term “prevent” or “prevention” means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease. 768698: NGT-003PC

[0105] As used herein, an “effective amount,” of a therapeutic agent disclosed herein is an amount sufficient to achieve a desired therapeutic or pharmacological effect, such as an amount that is capable of activating the growth of neurons. An effective amount of an agent as defined herein may vary according to factors such as the disease state, age, and weight of the subject, and the ability of the agent to elicit a desired response in the subject. Dosage regimens may be adjusted to provide the optimum therapeutic response. An effective amount is also one in which any toxic or detrimental effects of the active compound are outweighed by the therapeutically beneficial effects. As used herein, the term a “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutic result may be, e.g., lessening of symptoms, prolonged survival, improved mobility, and the like. A therapeutic result need not be a “cure.”

[0106] As used herein, the terms “improve,” “improving” or “improvement” or grammatical variations thereof used in relation to behaviors refer to the ability to achieve a measurable increase in performance in relation to tasks used to test these behaviors in a subject, including humans or non-human animals.

[0107] Central Nervous System

[0108] As used herein the term, “central nervous system (CNS) neurons” include the neurons of the brain, the cranial nerves, and the spinal cord.

[0109] As used herein, the term “neurological disorder” or “neurological condition” includes a disease, disorder, or condition which directly or indirectly affects the normal functioning or anatomy of a subject’s nervous system. The term “stroke” is art-recognized and includes sudden diminution or loss of consciousness, sensation and voluntary motion caused by rupture or obstruction (for example, by a blood clot) of an artery of the brain. “Traumatic brain injury” is art-recognized and includes the condition in which a traumatic blow to the head causes damage to the brain or connecting spinal cord, with or without penetrating the skull. Usually, the initial trauma can result in expanding hematoma, subarachnoid hemorrhage, cerebral edema, raised intracranial pressure, and cerebral hypoxia, which can, in turn, lead to severe secondary events due to low cerebral blood flow.

[0110] As used herein, the term axonal “growth” or “outgrowth” (also referred to herein as “neuronal outgrowth”) includes the process by which axons or dendrites extend from a neuron. The outgrowth can result in a new neuritic projection or in the extension of a previously existing cellular process. Axonal outgrowth may include linear extension of an axonal process by five cell-diameters or more. Neuronal growth processes, including neuritogenesis, can be evidenced by GAP-43 expression detected by methods such as immunostaining. “Stimulating axonal growth” means promoting axonal outgrowth. 768698: NGT-003PC

[0111] As used herein, the term “dieback” refers to axonal retraction that occurs as a result of trauma to the axon.

[0112] As used herein, the term “retraction” refers to the receding of the axon away from the site of injury, such as from where the glial scar forms. Here, the end of regenerating axons stops extending and become dystrophic. These dystrophic ends then can recede further from the glial scar and the site of injury.

[0113] Peptide Dimers

[0114] Receptor-type protein tyrosine phosphatases (PTPRs) are a subgroup of protein tyrosine phosphatases (PTPs) that have been identified as receptors for chondroitin sulfate proteoglycans (CSPGs), the principal inhibitory constituents of the glial scar and perineuronal net. Since CSPGs are the primary impediment to regeneration and plasticity in the injured adult nervous system, modulators of the LAR family protein tyrosine phosphatase functions can be used as therapeutic agents promoting neural plasticity, regeneration and ultimately repair of nervous system damage.

[0115] This application relates to peptidyl compounds that are useful for repairing the nervous system in a subject in need thereof.

[0116] Accordingly, in an aspect, provided herein is a peptide dimer of Formula (I):

[0117] (I), or a pharmaceutically acceptable salt thereof, wherein:

[0118] X1ais a first peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of TAT;

[0119] X2ais a second peptidyl domain comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR);

[0120] X1bis a third peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of TAT;

[0121] X2bis a fourth peptidyl domain comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR);

[0122] L is 768698: NGT-003PC

[0123] R is -C(i-6)alkylene- or -C(i-6)alkylene-[OCH2CH2]n-O-C(i-6)alkylene-; and n is 1-20.

[0124] In some embodiments, the present disclosure provides a peptide dimer of Formula (I):

[0125] (I), or a pharmaceutically acceptable salt thereof, wherein:

[0126] X1ais a first peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3;

[0127] X2ais a second peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15;

[0128] X1bis a third peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3;

[0129] X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15;

[0130] R is -C(i-6)alkylene- or -C(i-6)alkylene-[OCH2CH2]n-O-C(i-6)alkylene-; and n is 1-20.

[0131] In some embodiments, the present disclosure provides a peptide dimer of Formula (I): 768698: NGT-003PC

[0132] (I), or a pharmaceutically acceptable salt thereof, wherein:

[0133] X1ais a first peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3;

[0134] X2ais a second peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, and 15;

[0135] X1bis a third peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3;

[0136] X2bis a fourth peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, and 15;

[0137] R is -C(i-6)alkylene- or -C(i-2)alkylene-[OCH2CH2]n-O-C(i-2)alkylene-; and n is 1-20.

[0138] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (I’):

[0139] (I’), or a pharmaceutically acceptable salt thereof.

[0140] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (I”): 768698: NGT-003PC

[0141] (I”), or a pharmaceutically acceptable salt thereof.

[0142] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (I’”):

[0143] (I’”), or a pharmaceutically acceptable salt thereof.

[0144] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (I””):

[0145] (I””), or a pharmaceutically acceptable salt thereof.

[0146] In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of wild type HIV TAT.

[0147] In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence having at least 65% identity to wild type HIV TAT. In some embodiments, X1ais a 768698: NGT-003PC first peptidyl domain comprising an amino acid sequence having at least 70% identity to wild type HIV TAT. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence having at least 75% identity to wild type HIV TAT. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence having at least 80% identity to wild type HIV TAT. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence having at least 85% identity to wild type HIV TAT. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence having at least 90% identity to wild type HIV TAT. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence having at least 95% identity to wild type HIV TAT. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of wild type HIV TAT.

[0148] In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3 (depicted below in Table 1 ). In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID

[0149] NOS: 1 , 2, and 3.

[0150] Table 1. TAT Sequences

[0151] In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, 768698: NGT-003PC

[0152] X1ais a first peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1.

[0153] In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2.

[0154] In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID 768698: NGT-003PC

[0155] NO: 3. In some embodiments, X1ais a first peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3.

[0156] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 65% identity to a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR). In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 70% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 75% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 80% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 85% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 90% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 95% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2ais a second peptidyl domain comprising the amino acid sequence of a cytoplasmic wedge domain of a PTPR.

[0157] In some embodiments, the PTPR is a LAR family phosphatase. Structural and sequence analysis has revealed that all members of the LAR family contain a conserved 24 amino acid wedge-shaped helix-loop-helix motif in the first intracellular catalytic domain that can potentially mediate homo / heterophilic receptor interaction. Table 2 lists the amino acid sequences of intracellular portions of the LAR family phosphatase members that contain the wedge domain. The 24 amino acid wedge domains of these intracellular portions of LAR family phosphatases are identified by underlining. While the specific structure of the wedge domain is conserved through most LAR family wedge domains, the exact amino acids that make up the wedge domains vary between individual proteins and sub-families.

[0158] As can be seen in Table 2, the wedge domain is highly conserved across members of the LAR family. For example, the wedge domain sequence of PTPRS is highly conserved among mammals, with only a single amino acid change in mice and rats (Threonine to Methionine at position 6). 768698: NGT-003PC

[0159] Table 2: Wedge Domain of LAR Family Phosphatase

[0160] Accordingly, in some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to about 10 consecutive amino acids of the wedge domain of a LAR family phosphatase. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to about 15 consecutive amino acids of the wedge domain of a LAR family phosphatase. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to about 20 consecutive amino acids of the wedge domain of a LAR family phosphatase.

[0161] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to 10-20 consecutive amino acids of a wedge domain of PTPRF, PTPRD, or PTPRS.

[0162] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence derived from a PTPRF wedge domain, a PTPRD wedge domain, and a PTPRS wedge domain, and variants having at least 65% identity thereto, at least 70% identity thereto, at least 75% identity thereto, at least 80% identity thereto, at least 85% identity thereto, at least 90% identity thereto, or at least 95% identity thereto.

[0163] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15 (depicted below in Table 3). In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, 768698: NGT-003PC

[0164] X2ais a second peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence selected from the group consisting of any one of SEQ ID NOs: 12, 13, 14, or 15.

[0165] Table 3: Wedge domain sequence of LAR family phosphatases

[0166] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12.

[0167] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence 768698: NGT-003PC that is at least 70% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13.

[0168] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14.

[0169] In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl 768698: NGT-003PC domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0170] In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of wild type HIV TAT.

[0171] In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 65% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 70% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 75% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 80% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 85% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 90% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence having at least 95% identity to wild type HIV TAT. In some embodiments, X1bis a third peptidyl domain comprising the amino acid sequence of wild type HIV TAT.

[0172] In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3.

[0173] In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 1 . In 768698: NGT-003PC some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 1. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1 . In some embodiments, X1bis a third peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1.

[0174] In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 2. In some embodiments, X1bis a third peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2.

[0175] In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid 768698: NGT-003PC sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, X1bis a third peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3.

[0176] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 65% identity to a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR). In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 70% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 75% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 80% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 85% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 90% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence having at least 95% identity to a cytoplasmic wedge domain of a PTPR. In some embodiments, X2bis a fourth peptidyl domain comprising the amino acid sequence of a cytoplasmic wedge domain of a PTPR.

[0177] In some embodiments, X2bis a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to about 10 consecutive amino acids of the wedge domain of a LAR family phosphatase. In some embodiments, X2bis a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to about 15 consecutive amino acids of the wedge domain of a LAR family phosphatase. In some embodiments, X2bis a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to about 20 consecutive amino acids of the wedge domain of a LAR family phosphatase.

[0178] In some embodiments, X2bis a second peptidyl domain comprising an amino acid sequence having at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identity to 10-20 consecutive amino acids of a wedge domain of PTPRF, PTPRD, or PTPRS. 768698: NGT-003PC

[0179] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence derived from a PTPRF wedge domain, a PTPRD wedge domain, and a PTPRS wedge domain, and variants having at least 65% identity thereto, at least 70% identity thereto, at least 75% identity thereto, at least 80% identity thereto, at least 85% identity thereto, at least 90% identity thereto, or at least 95% identity thereto.

[0180] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, or 15.

[0181] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, X2bis a fourth peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12. 768698: NGT-003PC

[0182] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 13. In some embodiments, X2bis a fourth peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13.

[0183] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, X2bis a fourth peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14.

[0184] In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 65% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 80% 768698: NGT-003PC identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 85% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 15. In some embodiments, X2bis a fourth peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0185] In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0186] In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0187] In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1ais a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2ais a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0188] In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2bis a second peptidyl domain comprising the amino acid 768698: NGT-003PC sequence of SEQ ID NO: 12. In some embodiments, X1 bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 1 , and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0189] In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, X1 bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 2, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0190] In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 12. In some embodiments, X1 bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 13. In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, X1bis a first peptidyl domain comprising the amino acid sequence of SEQ ID NO: 3, and X2bis a second peptidyl domain comprising the amino acid sequence of SEQ ID NO: 15.

[0191] In some embodiments, X1aand X1bare identical. In some embodiments, X1aand X1bare different.

[0192] In some embodiments, X2aand X2bare identical. In some embodiments, X2aand X2bare different.

[0193] In some embodiments, the peptide dimer may include additional residues at the C- terminus or the N-terminus of one or both peptide monomers. In some embodiments, the peptide may include a peptide tag at the C-terminus or the N-terminus of one or both peptide monomers. In some embodiments, the tag may be an affinity tag such as a His-tag, Flag Tag, or Twin-Strep Tag.

[0194] In some embodiments, the peptide further comprises an N-terminal capping group. Accordingly, in some embodiments, the first peptidyl domain further comprises an N-terminal capping group. In some embodiments, the third peptidyl domain further comprises an N- 768698: NGT-003PC terminal capping group. In some embodiments, the N-terminal capping group is an acetyl group (Ac). In some embodiments, the peptide comprises amino groups at the N-termini and does not comprise an N-terminal capping group.

[0195] In some embodiments, the peptide further comprises a C-terminal capping group. Accordingly, in some embodiments, the second peptidyl domain further comprises a C- terminal capping group. In some embodiments, the fourth peptidyl domain further comprises a C-terminal capping group. Nonlimiting examples of C-terminal capping groups include - NH2, -NHMe, -NHEt, -OMe, -OEt, and -OtBu. In some embodiments, the peptide is amidated at the C-termini and comprises C-terminal capping groups that are -NH2. In some embodiments, the peptide comprises carboxylic acids at the C-termini and does not comprise a C-terminal capping group.

[0196] Peptide dimers described herein may also include, for example, biologically active mutants, variants, fragments, chimeras, and analogues. The term "fragments” encompasses amino acid sequences having truncations of one or more amino acids from the amino terminus (N-terminus), the carboxy terminus (C-terminus), or the interior of the peptide. Analogues of the peptide dimers of the present disclosure may include an insertion or a substitution of one or more amino acids. Variants, mutants, fragments, chimeras and analogues may function as inhibitors to abolish inhibitory effects of CSPGs on neural cells activated with CSPGs (without being restricted to the present examples).

[0197] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (la):

[0198] H II N GRKKRRQRRR" ^ DMAEHTERLKANDSLKLSQEYESI-NH2

[0199] I L I GRKKRRQRRR .NS / 1k / DMAEHTERLKANDSLKLSQEYESI-NH2H I

[0200] (la), or a pharmaceutically acceptable salt thereof.

[0201] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (la’): 768698: NGT-003PC

[0202] H H „N. GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH2

[0203] I L I

[0204] GRKKRRQRRR . DMAEHTERLKANDSLKLSQEYESI-NH2

[0205] (la’), or a pharmaceutically acceptable salt thereof.

[0206] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (la”):

[0207] GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH 2

[0208] GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH2

[0209] (la”), or a pharmaceutically acceptable salt thereof.

[0210] In some embodiments, the peptide dimer of Formula (I) has a structure according to

[0211] Formula (la’”):

[0212] (la’”), or a pharmaceutically acceptable salt thereof.

[0213] In some embodiments, the peptide dimer of Formula (I) has a structure according to Formula (la””): 768698: NGT-003PC

[0214] H H

[0215] GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH2

[0216] GRKKRRQRRR .

[0217] N DMAEHTERLKANDSLKLSQEYESI-NH2

[0218] H(5

[0219] (la””), or a pharmaceutically acceptable salt thereof.

[0220] In some embodiments, R is -C(i.6)alkylene-. In some embodiments, R is -CH2-. In some embodiments, R is -CH2CH2-. In some embodiments, R is -CH2CH2CH2-. In some embodiments, R is -CH2CH2CH2CH2-. In some embodiments, R is -CH2CH2CH2CH2CH2-. In some embodiments, R is -CH2CH2CH2CH2CH2CH2-.

[0221] In some embodiments, R is -C(i.6)alkylene-[OCH2CH2]n-O-C(i.6)alkylene-. In some embodiments, R is -C(i.2)alkylene-[OCH2CH2]n-O-C(i.2)alkylene-. In some embodiments, R is -CH2-[OCH2CH2]n-OCH2-. In some embodiments, R is -CH2CH2-[OCH2CH2]n-OCH2CH2-.

[0222] In some embodiments, n is 1 -3. In some embodiments, n is 1 -6. In some embodiments, n is 1-10. In some embodiments, n is 10-20. In some embodiments, n is 1 . In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, n is 11 . In some embodiments, n is 12. In some embodiments, n is 13.

[0223] In some embodiments, n is 14. In some embodiments, n is 15. In some embodiments, n is 16. In some embodiments, n is 17. In some embodiments, n is 18. In some embodiments, n is 19. In some embodiments, n is 20.

[0224] In some embodiments, R is -CH2CH2OCH2CH2OCH2CH2-.

[0225] The present disclosure further provides a peptide dimer having the following structure:

[0226] 768698: NGT-003PC

[0227] DMAEHTERLKANDSLKLSQEYESI-NH2

[0228] GRKKRR ERLKANDSLKLSQEYESI-NH2 or a pharmaceutically acceptable salt thereof.

[0229] The present disclosure further provides a peptide dimer having the following structure: or a pharmaceutically acceptable salt thereof.

[0230] In various nonlimiting embodiments, the peptides disclosed herein may be used as therapeutic agents to promote cell growth, motility, survival and plasticity of these cells.

[0231] Preparation of the Peptide Dimers The present disclosure further provides methods of preparing peptide monomers used for preparing the peptide dimers and methods of preparing the peptide dimers disclosed herein.

[0232] According to various embodiments, a peptide monomer used to prepare the peptide dimers may be prepared by methods known to those skilled in the art. For example, a 768698: NGT-003PC peptide monomer can be prepared using conventional peptide synthesis techniques (e.g., solid phase synthesis) or molecular biology techniques.

[0233] In an embodiment, a peptide monomer used for forming a peptide dimer disclosed herein may be produced by genetic engineering using a recombinant DNA. For example, a recombinant DNA may be engineered to encode a fusion peptide used for making a peptide dimer disclosed herein. The fusion peptide may comprise a peptide domain comprising an amino acid sequence selected from leukocyte antigen related (LAR) family phosphatase wedge domains or variants having at least 65% homologous thereto, and the peptide domain may be connected to a peptidyl domain comprising an amino acid sequence derived from a PTD of TAT, as disclosed herein, via a cysteine residue.

[0234] This recombinant DNA may be inserted in an expression cassette of an expression vector and operably linked to a regulatory region. The regulatory region typically comprises a promoter to regulate the expression of the peptide monomer in a cell carrying the vector. In yet another embodiment, the promoter is a constitutive promoter, such as a CMV, such that the peptide monomer may be expressed consistently in a cell carrying the vector. In still another embodiment, the promoter is an inducible promoter, and the expression of the peptide monomer can be induced as needed.

[0235] In an embodiment, the vector is a plasmid vector and can be transformed into bacteria to store or to amplify, and can be transfected into mammalian cells to express the recombinant peptide.

[0236] In another embodiment, the preparation disclosed herein can include cultivating a host cell (bacterial or eukaryotic) under conditions, which provide for the expression of peptides and / or proteins within the cell.

[0237] The peptide monomer expressed in the host cell can be purified by affinity methods, ion exchange chromatography, size exclusion chromatography, hydrophobicity or other purification technique typically used for protein purification. The purification step can be performed under non-denaturing conditions. On the other hand, if a denaturating step is required, the protein may be renatured using techniques known in the art.

[0238] In yet another embodiment, the peptide monomers described herein can include additional residues that may be added at either terminus of a polypeptide for the purpose of providing a “linker” by which the polypeptides can be conveniently linked and / or affixed to other polypeptides, proteins, detectable moieties, labels, solid matrices, or carriers.

[0239] Amino acid residue linkers are usually at least one residue and can be 40 or more residues, more often 1 to 10 residues. Typical amino acid residues used for linking are glycine, tyrosine, cysteine, lysine, glutamic and aspartic acid, or the like. In addition, a subject polypeptide can differ by the sequence being modified by terminal-NH2 acylation, e.g., acetylation, or thioglycolic acid amidation, by terminal-carboxylamidation, e.g., with 768698: NGT-003PC ammonia, methylamine, and the like terminal modifications. Terminal modifications are useful, as is well known, to reduce susceptibility by proteinase digestion, and therefore serve to prolong half life of the polypeptides in solutions, particularly biological fluids where proteases may be present. In this regard, polypeptide cyclization is also a useful terminal modification, and is particularly preferred also because of the stable structures formed by cyclization and in view of the biological activities observed for such cyclic peptides as described herein.

[0240] In still another embodiment, the linker can be a flexible peptide linker that links the therapeutic peptide to other polypeptides, proteins, and / or molecules, such as detectable moieties, labels, solid matrices, or carriers. A flexible peptide linker can be about 20 or fewer amino acids in length. For example, a peptide linker can contain about 12 or fewer amino acid residues, e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11 , and 12. In some cases, a peptide linker comprises two or more of the following amino acids: glycine, serine, alanine, and threonine.

[0241] A peptide dimer disclosed herein may be prepared by dimerization of peptide monomers as prepared above, where the peptide monomers form the monomer subunits in the peptide dimer. The peptide monomers used to prepare a dimer may be identical or different, resulting in a homodimer or heterodimer, respectively.

[0242] Formulations

[0243] The present disclosure further provides a pharmaceutical composition comprising a peptide dimer described herein and a pharmaceutically acceptable carrier.

[0244] Nonlimiting examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers; alumina; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate; disodium hydrogen phosphate; potassium hydrogen phosphate; sodium chloride; zinc salts; colloidal silica; magnesium trisilicate; polyvinyl pyrrolidone; polyacrylates; waxes; polyethylenepolyoxypropylene-block polymers; wool fat; sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such a propylene glycol or polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; and phosphate buffer solutions. 768698: NGT-003PC

[0245] Further, non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator.

[0246] According to various embodiments, the pharmaceutical composition may be formulated in any suitable form for delivery to a subject in need thereof, either with fixed- dose or non-fixed dose. For example, the pharmaceutical composition may be adapted for oral or parenteral and may be administered to the subject in the dosage form of tablets, sugar-coated tablets, capsules, delayed- release hard capsules, softgel, chewable tablets, gummy, caplets, powders, granules, syrups, aerosols, inhalants, suppositories, solutions, suspensions, catheters containing the composition, syringes containing the composition, implants containing the composition, transdermal patch, or the like.

[0247] In some embodiments, the pharmaceutical compositions are formulated in liquid solution, typically in physiologically compatible buffers such as Hank’s solution or Ringer’s solution for injection. In another embodiment, a therapeutic agent comprising a peptide dimer or pharmaceutically acceptable analog, salt, or solvate thereof described herein may be formulated in solid form and re-dissolved or suspended immediately in a pharmaceutical acceptable solvent prior to use. For example, a peptide dimer or pharmaceutically acceptable analog, salt, or solvate thereof may be in lyophilized form that can be dissolved to obtain a final preparation to administer to a subject at the time of use. Injectable preparations (for example, sterile injectable aqueous or oleaginous suspensions) may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s solution, U.S.P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0248] In some embodiments, the pharmaceutical composition does not comprise DMSO.

[0249] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous (s.c.) or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution which, in turn, may depend upon crystal size and crystalline form. Absorption of a parenterally administered drug form can also be delayed by dissolving or suspending the drug in an oil vehicle. 768698: NGT-003PC

[0250] In still other embodiments, the pharmaceutical compositions disclosed herein may be formulated in a dosage form such as a tablet, a softgel, a capsule, a caplet, a polypill, a chewable tablet, a gummy, a hard capsule, a transdermal patch, etc.

[0251] In some embodiments, the pharmaceutical compositions disclosed herein may be formulated in liquid form for oral administration, where the pharmaceutical composition includes, among with the therapeutic agents (or active compounds), pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, com, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0252] Solid compositions of a similar type may also be employed as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0253] Active ingredients comprising the peptide dimers or pharmaceutically acceptable analog, salt, or solvate thereof can also be in micro-encapsulated form with one or more excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings, and other coatings well known in the pharmaceutical formulating art. In such solid dosage forms the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents.

[0254] The pharmaceutical compositions may be formulated in forms for topical or transdermal administration, where the forms may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, eye ointments, powders and solutions are also contemplated as being within the scope of this disclosure. 768698: NGT-003PC

[0255] The ointments, pastes, creams and gels may contain, in addition to an active compound of this disclosure, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0256] Powders and sprays can contain, in addition to the compounds of this disclosure, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.

[0257] Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0258] Methods of Treatment

[0259] The peptide dimers of the present disclosure, when being delivered to a cell (such as neural cells), are able to promote cell growth, motility, survival and plasticity in the cells, e.g., neural cells.

[0260] As such, the peptide dimers described herein can be used to abolish inhibitory effects of CSPGs on neural cells activated with CSPGs and promote cell growth, motility, and survival, and to treat diseases, disorders, and / or conditions associated with accumulation of CSPGs or with activation and signaling of LAR family of phosphatases.

[0261] Accordingly, the present disclosure provides methods for treating diseases, disorders, and / or conditions associated with accumulation of CSPGs or with activation and signaling of LAR family of phosphatases using the peptide dimers or compositions comprising the peptide dimers disclosed herein.

[0262] One potential mechanism for regulation, modulation, and / or inhibition of LAR family of phosphatases involves dimerization of the intracellular portion of the phosphatase. In contrast to receptor tyrosine kinases, which are active as dimers and inactive as monomers, several protein tyrosine phosphatases (PTPs) have been shown to be inactive as dimers and active as monomers. PTPalpha, PTP1 B and CD45, which have been crystalized in both forms, have been shown to be active as monomers and inactive as dimers. As PTPRF demonstrates homophillic binding under specific oxidative conditions, and PTPRS can dimerize in response to ligand binding, it is suggested that ligands to LAR family phosphatases can direct the activation state of PTPRF and PTPRS. Therefore, mimicking dimerization with intracellular-targeted therapies may directly inactivate LAR family of phosphatases without alteration of the extracellular matrix or other ligands. 768698: NGT-003PC

[0263] Peptide mimetics of the intracellular portion of the LAR family of phosphatase, when delivered into a neural cell, may inhibit and / or reduce LAR activity or function induced by CSPG. Suppression of LAR family activity, signaling, and / or function in response to CSPG activation was found to promote neural cell outgrowth, including restoration of growth cone motility, extension of processes, sprouting, promotion of neural cell survival and plasticity, and inhibit neural cell dieback.

[0264] Thus, therapeutic peptides that bind to and / or complex with the intracellular domain of at least one LAR family phosphatase, such as the peptide dimers of the present disclosure, can be used to promote cell growth, motility, survival and plasticity of neural cells. Accordingly, in some embodiments, the function of a LAR family phosphatase is inhibited or reduced by a peptide dimer described herein. In other embodiments, one or more of the activities and signaling of the LAR family phosphatase is inhibited or reduced by a peptide dimer described herein.

[0265] In some embodiments, provided herein is a method for repairing the nervous system of a subject in need thereof comprising administering to said subject an effective amount of a peptide dimer described herein.

[0266] In various embodiments, the present disclosure provides a method of treating a neurological condition, disease or disorder selected from the group consisting of neural injury, neurological disease caused by inflammation or autoimmunity, neurodegene rati ve disease, and a neurological condition, in a subject in need thereof, the method comprising administering an effective amount of a peptide dimer disclosed herein to the subject.

[0267] In an embodiment, the neural injury is selected from the group consisting of acute neural injury, traumatic brain injury (TBI), spinal cord injury, concussion, stroke, including ischemic stroke, hemorrhagic stroke, and chronic stroke disease.

[0268] In another embodiment, the neurological condition, disease or disorder is selected from the group consisting of Alzheimer’s Disease, dementias related to Alzheimer’s Disease, Lewy diffuse body diseases, senile dementia, Parkinson’s Disease, amyotrophic lateral sclerosis, multiple sclerosis (MS), optic neuritis, Huntington’s Disease, Tourette’s syndrome, hereditary motor and sensory neuropathy, diabetic neuropathy, progressive supranuclear palsy, Jakob-Creutzfeldt disease, epilepsy, and infectious disease.

[0269] Various embodiments further provide a use of a peptide dimer disclosed herein in the manufacture of a medicament for treatment of a neurological condition, disease or disorder selected from the group consisting of neural injury, neurological disease caused by inflammation or autoimmunity, neurodegenerative disease, a neurological condition, or a combination thereof. 768698: NGT-003PC

[0270] Administration and Dosages

[0271] In general, the peptide dimers disclosed herein may be administered to a subject in need thereof via any suitable route, including, for example, orally (e.g., in capsules, suspensions or tablets), systemically, or by parenteral administration. Non-limiting example routes include subcutaneous, intramuscular, intravenous, transdermal, intranasal, rectal, ocular, topical, sublingual, and buccal.

[0272] In an embodiment, the peptide dimers can be administered by lateral cerebroventricular injection into the brain of a subject, usually within 100 hours of when an injury (resulting in a condition characterized by aberrant axonal outgrowth of central nervous system neurons) occurs (such as within 6, 12, 24 or 100 hours, inclusive, from the time of the injury). The injection can be made, for example, through a burr hole made in the subject’s skull. In another embodiment, the therapeutic agent can be administered through a surgically inserted shunt into the cerebral ventricle of a subject, usually within 100 hours of when an injury occurs (e.g., within 6, 12 or 24 hours, inclusive, from the time of the injury). For example, the injection can be made into the lateral ventricles, which are larger, even though injection into the third and fourth smaller ventricles can also be made. In yet another embodiment, the therapeutic agent can be administered by injection into the cisterna magna, or lumbar area of a subject, within 100 hours of when an injury occurs (such as within 6, 12, or 24 hours, inclusive, from the time of the injury).

[0273] In still another embodiment, the peptide dimer can be administered to a subject at or near the site of injury, usually within 100 hours of when an injury occurs (e.g., within 6, 12, or 24 hours, inclusive, of the time of the injury). Such administration may optionally be subcutaneous.

[0274] In an embodiment, the pharmaceutical compositions can be administered to a subject 100 hours or more after the time of injury. In some cases, the administration will be a week after injury, multiple weeks after injury, or months or years after injury.

[0275] In another embodiment, a therapeutic amount or dose of the peptide dimers disclosed herein may range from about 0.1 mg / Kg to about 500 mg / Kg per body weight. In yet another embodiment, a therapeutic amount or dose of the peptide dimers disclosed herein may range from about 1 to about 50 mg / Kg. In general, treatment regimens according to the present disclosure comprise administration to a subject in need of such treatment from about 10 mg to about 1000 mg of the peptide dimers of this disclosure per day in single or multiple doses. Therapeutic amounts or doses will also vary depending on route of administration, as well as the possibility of co-usage with other agents.

[0276] It will be understood, however, that the total daily usage of the compounds and compositions of the present disclosure will be decided by the attending physician within the scope of sound medical judgment. The specific inhibitory dose for any particular subject will 768698: NGT-003PC depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific therapeutic agents or compositions employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.

[0277] Kits

[0278] One aspect of the present disclosure relates to a kit comprising a therapeutic agent comprising a peptide dimer or pharmaceutically acceptable salt thereof, or any composition comprising the peptide dimer or pharmaceutically acceptable salt thereof described herein.

[0279] In an embodiment, the kit comprises one or more separate dosage forms, each dosage form comprises a pharmaceutical composition comprising an effective amount or one dose of a peptide dimer or pharmaceutically acceptable salt thereof disclosed herein for treating diseases, disorders, and / or conditions associated with inhibition of nervous system repair by chondroitin sulfate proteoglycans (CSPG) or with suppressive effects of LAR family phosphatases, such as PTPRD, PTPRF, and PTPRS on neuro repair .

[0280] In another embodiment, the kit may comprise a first container comprising a peptide dimer or pharmaceutically acceptable salt thereof according to the present disclosure that is in free form, such as lyophilized powder, and optionally, a second container comprising a pharmaceutically acceptable solvent to dissolve the peptide dimer. At the time of use, the peptide dimer in free form can be mixed with the solvent to produce a final preparation for administering to a subject in need thereof.

[0281] In yet another embodiment, the kit may further include an instruction for using the therapeutic agent or composition comprised in the kit for treating diseases, disorders, and / or conditions associated with inhibition of nervous system repair by chondroitin sulfate proteoglycans (CSPG) or with suppressive effects of LAR family phosphatases, such as PTPRD, PTPRF, and PTPRS on neuro repair.

[0282] It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present disclosure. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

[0283] The embodiments described herein are not limited to the particular methodology, protocols, and reagents, etc., and as such may vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting in 768698: NGT-003PC scope. Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term “about.”

[0284] All patents and other publications identified are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the present disclosure. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior disclosure or for any other reason. All statements as to the date or representation as to the contents of these documents are based on the information available to the applicants and do not constitute any admission as to the correctness of the dates or contents of these documents.

[0285] The following examples further illustrate aspects of the present disclosure. However, they are in no way a limitation of the teachings of the present disclosure as set forth. It should be understood that these Examples are given by way of illustration only. From the above discussion and these Examples, one of ordinary skill in the art can ascertain the essential characteristics of embodiments of the present disclosure. Without departing from the spirit and scope thereof, one skilled in the art can make various changes and modifications of the disclosure to adapt it to various usages and conditions. All publications, including patents and non-patent literature, referred to in this specification are expressly incorporated by reference herein.

[0286] EXAMPLES

[0287] The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of organic synthesis, cell biology, cell culture, and molecular biology, which are within the skill of the art.

[0288] Example 1 : Synthesis and purification of peptide dimers

[0289] Raw Materials

[0290] Compound 1 (peptide monomer comprising amino acid sequence of TAT-Cys-human

[0291] PTPRS wedge domain: GRKKRRQRRRCDMAEHTERLKANDSLKLSQEYESI (SEQ ID NO: 16).

[0292] Purified water for purification (PWP)

[0293] Buffer Components:

[0294] Ethanol (EtOH)

[0295] Acetic Acid (AcOH)

[0296] Acetonitrile (ACN)

[0297] Methanol (MeOH) 768698: NGT-003PC

[0298] Ammonium Acetate (NH4OAc)

[0299] Linkers: bismaleimidoethane (BMOE)

[0300] 1 ,8-bismaleimido-diethyleneglycol (BM(PEG)2)

[0301] Compound 2: [H-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-11Cys-Asp-Met-Ala-Glu-His-Thr- Glu-Arg-Leu-Lys-Ala-Asn-Asp-Ser-Leu-Lys-Leu-Ser-Gln-Glu-Tyr-Glu-Ser-lle-NH2]2(SEQ ID NO: 16) (11Cys-11Cys linked with BMOE)

[0302] H H „N. GRKKRRQRRR DMAEHTERLKANDSLKLSQEYESI-NH2

[0303] GRKKRRQRR HTERLKANDSLKLSQEYESI-NH2

[0304] Preparation and purification of Compound 2 - Route A

[0305] Compound 1 was reacted with linker BMOE. The resulting crude Compound 2 was purified by preparatory HPLC using a C8 column with 0.1 % AcOH as Mobile Phase A and 100% EtOH as Mobile Phase B using a gradient program. After the first purification, the material was subjected to a second purification to improve the purity. This purification employed a preparatory HPLC and a C8 column but with 0.2% TFA as Mobile Phase A and MPA + ACN (1 : 1 ) as Mobile Phase B using a gradient program. Purified fractions were lyophilized, resuspended in 0.1 N AcOH for subsequent Dowex® ion exchange in 0.1 N AcOH in batch mode. After Dowex® ion exchange, the solution was filtered, AcOH added to the solution (so that the final volume contained 10% AcOH) and lyophilized to obtain the final product in the form of an acetate salt. MW: 8797.85. Purity >95% (HPLC).

[0306] Preparation and purification of Compound 2 - Route B

[0307] Compound 1 was reacted with linker BMOE. The resulting crude Compound 2 was purified by preparatory HPLC using a C4 column with 0.025% TFA as Mobile Phase A and 100% ACN as Mobile Phase B using a gradient program. Ion exchange & final purification was performed on a C4 column. The column was washed with wash 1 (95% of 0.1% AcOH + 5% of 100% ACN), wash 2 (95% of 0.1 M ammonium acetate + 5% of 100% ACN), wash 3 (95% of 0.1 % AcOH + 5% of 100% ACN), and the peptide was eluted using a gradient 768698: NGT-003PC program with 0.1 % AcOH +100% ACN as mobile phases. The purified material was lyophilized to obtain the final product in the form of an acetate salt. MW: 8797.85. Purity

[0308] Preparation and purification of Compound 3 - Route A

[0309] Compound 1 was reacted with linker BM(PEG)2. The resulting crude Compound 3 was purified by preparatory HPLC using a C8 column with 0.1% AcOH as Mobile Phase A and 100% EtOH as Mobile Phase B using a gradient program. Purified fractions were lyophilized to obtain the final product in the form of an acetate salt. MW: 8885.96. Purity >95% (HPLC).

[0310] Preparation and purification of Compound 3 - Route B

[0311] Compound 1 was reacted with linker BM(PEG)2obtained from ThermoFisher. The resulting crude Compound 3 was purified by preparatory HPLC using a C4 column with 0.025% TFA as Mobile Phase A and 100% ACN as Mobile Phase B using a gradient program. Ion exchange & final purification was performed on a C4 column. The column was washed with wash 1 (95% of 0.1 % AcOH + 5% of 100% ACN), wash 2 (95% of 0.1 M ammonium acetate + 5% of 100% ACN), wash 3 (95% of 0.1% AcOH + 5% of 100% ACN), and the peptide was eluted using a gradient program with 0.1 % AcOH +100% ACN as mobile phases. The purified material was lyophilized to obtain the final product in the form of an acetate salt. MW: 8885.96. Purity >95% (HPLC).

[0312] Example 2 - Distinct colloidal behavior of Compounds 2 and 3 768698: NGT-003PC

[0313] The patterns of self-assembly of Compounds 2 and 3 were analyzed by negative stain transmission electron microscopy.

[0314] Materials And Methods

[0315] Compound 2 and Compound 3 were dissolved in sterile water or in 0.9% (w / v) sodium chloride in sterile water to obtain a 20 mg / mL solution. Formvar / carbon coated grids were exposed to 15 mL drops of the Compound 2 or Compound 3 solutions for approximately 1 minute, blotted and then exposed to 15 mL drops of sterile water for 10 seconds, blotted again and then stained with 2% (w / v) aqueous uranyl acetate for 30 seconds. The stained samples were exposed to 15 mL drops of sterile water for 10 seconds and blotted dry prior to examination by transmission electron microscopy.

[0316] Results

[0317] FIG. 1 presents a set of transmission electron microscopy (TEM) images showing colloidal structures resulting from self-assembly of Compound 2 (panels “A” and “B”) and Compound 3 (panels “C” and “D”) in water (panels “A” and “C”) and in isotonic saline (panels “B” and “D”). The images of the compounds in water demonstrate formation of clumps by both Compound 2 and Compound 3. The images presented in the saline panels demonstrate the formation of an amyloid-like fibrillar meshwork by both compounds indicating that they have similar physicochemical behaviors in the simulated biological matrix (isotonic saline).

Claims

768698: NGT-003PCCLAIMS1. A peptide dimer of Formula (I):(I), or a pharmaceutically acceptable salt thereof, wherein:X1ais a first peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of TAT;X2ais a second peptidyl domain comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR);X1bis a third peptidyl domain comprising an amino acid sequence derived from a protein transduction domain (PTD) of TAT;X2bis a fourth peptidyl domain comprising an amino acid sequence derived from a cytoplasmic wedge domain of a receptor-type protein-tyrosine phosphatase (PTPR);L isR is -C(i-6)alkylene- or -C(i-6)alkylene-[OCH2CH2]n-O-C(i-6)alkylene-; and n is 1-20.

2. The peptide dimer of claim 1 , or a pharmaceutically acceptable salt thereof, wherein the first peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3.

3. The peptide dimer of claim 1 , or a pharmaceutically acceptable salt thereof, wherein the first peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3.768698: NGT-003PC4. The peptide dimer of claim 1 , or a pharmaceutically acceptable salt thereof, wherein the first peptidyl domain comprises the amino acid sequence of SEQ ID NO: 1 .

5. The peptide dimer of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, wherein the second peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15.

6. The peptide dimer of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, wherein the second peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, and 15.

7. The peptide dimer of any one of claims 1-4, or a pharmaceutically acceptable salt thereof, wherein the second peptidyl domain comprises the amino acid sequence of SEQ ID NO: 14.

8. The peptide dimer of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein the third peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 1 , 2, or 3.

9. The peptide dimer of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein the third peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 , 2, and 3.

10. The peptide dimer of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein the third peptidyl domain comprises the amino acid sequence of SEQ ID NO: 1.11 . The peptide dimer of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein the fourth peptidyl domain comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of any one of SEQ ID NOs: 12, 13, 14, or 15.

12. The peptide dimer of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein the fourth peptidyl domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 13, 14, and 15.768698: NGT-003PC13. The peptide dimer of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein the fourth peptidyl domain comprises the amino acid sequence of SEQ ID NO: 14.

14. The peptide dimer of claim 1 , or a pharmaceutically acceptable salt thereof, having a structure according to Formula (la): DMAEHTERLKANDSLKLSQEYESI-NH2GRKKRRQRRR ^NS_1A .DMAEHTERLKANDSLKLSQEYESI-NH2HI(la).

15. The peptide dimer of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein R is -C(i.6)alkylene-.

16. The peptide dimer of any one of claims 1-15, or a pharmaceutically acceptable salt thereof, wherein R is -CH2CH2-.

17. The peptide dimer of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein:R is -C(i-2)alkylene-[OCH2CH2]n-O-C(i-2)alkylene-; and n is 1-6.

18. The peptide dimer of any one of claims 1-14 and 17, or a pharmaceutically acceptable salt thereof, wherein R is -CH2CH2OCH2CH2OCH2CH2-.

19. The peptide dimer of claim 1 , or a pharmaceutically acceptable salt thereof, having a structure selected from the group consisting of:768698: NGT-003PCH H „l\k20. A pharmaceutical composition comprising the peptide dimer of any one of claims 1 - 19, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.21 . A method of repairing the nervous system and / or treating a neurological condition, disease or disorder selected from the group consisting of neural injury, neurological disease caused by inflammation or autoimmunity, and neurodegenerative disease in a subject in need thereof, the method comprising administering an effective amount of the pharmaceutical composition of claim 20 to the subject.

22. The method of claim 21 , wherein the neural injury is selected from the group consisting of acute neural injury, traumatic brain injury (TBI), spinal cord injury, concussion,768698: NGT-003PC stroke, including ischemic stroke, hemorrhagic stroke, chronic stroke disease, aneurysm, cerebral hemorrhage, thrombus, and embolism.

23. The method of claim 21 , wherein the neurological condition, disease or disorder is selected from the group consisting of Alzheimer’s Disease, dementias related to Alzheimer’s Disease, Lewy diffuse body diseases, senile dementia, Parkinson’s Disease, amyotrophic lateral sclerosis, multiple sclerosis (MS), optic neuritis, Huntington’s Disease, Tourette’s syndrome, hereditary motor and sensory neuropathy, diabetic neuropathy, progressive supranuclear palsy, Jakob-Creutzfeldt disease, epilepsy, and infectious disease.

24. Use of the pharmaceutical composition of claim 20 in the manufacture of a medicament for treatment of a neurological condition, disease or disorder selected from the group consisting of neural injury, neurological disease caused by inflammation or autoimmunity, and neurodegenerative disease.

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