WO2018140563A1

WO2018140563A1 - Use of apoe mimetic peptides for reducing the likelihood of epileptogenesis

Info

Publication number: WO2018140563A1
Application number: PCT/US2018/015165
Authority: WO
Inventors: Daniel T. Laskowitz; Brian E. MACE; Bradley Kolls
Original assignee: Duke University
Priority date: 2017-01-26
Filing date: 2018-01-25
Publication date: 2018-08-02

Abstract

Provided herein is a method for reducing the likelihood of epileptogenesis in a subject (e.g., a subject at risk thereof), comprising administering to said subject (e.g., a human subject) an effective amount a peptide of Formula I, or a pharmaceutically acceptable salt thereof.

Description

USE OF APOE MIMETIC PEPTIDES FOR REDUCING THE

LIKELIHOOD OF EPILEPTOGENESIS

BACKGROUND

Following moderate or severe brain injury, post-traumatic seizures are relatively common. Conventional anti-seizure medications are effective in suppressing early seizures, defined as events occurring within the first seven days of injury. However, suppression of early seizures does not reduce the incidence of the development of late seizures, known as epileptogenesis, in which seizures can occur weeks, months, or even years after the injury and are often associated thereafter with a lifetime burden of epilepsy.

Unfortunately, the mechanisms by which epileptogenesis occurs remain incompletely defined. At present, there are no drugs on the market that prevent the development of epilepsy after injury, as opposed to anti-epileptic drugs that suppress seizures once an individual has epilepsy. Thus, treatment of epileptogenesis remains an unmet medical challenge. See, e.g. , Goldberg and Coulter, Mechanisms of epileptogenesis: a convergence on neural circuit dysfunction, Nat. Rev. Neurosci. vol. 14(5), 337-349 (2013); Pitkanen and Lukasiuk, Review: Mechanisms of epileptogenesis and potential treatment targets, The Lancet Neurology vol. 10, 173-186 (2011).

US Patent No. 7,501 ,429 to Weaver also notes that, while a variety of drugs are available for the management of epileptic seizures, including older anticonvulsant agents such as phenytoin, valproate and carbamazepine, as well as newer agents like felbamate, gabapentin, and tiagabine, these drugs do not prevent epileptogenesis.

Thus, despite the numerous drugs available for the treatment of epilepsy through suppression of the convulsions associated with epileptic seizures, there are no generally accepted drugs for the treatment of the pathological changes which characterize epileptogenesis. There is no generally accepted method of inhibiting the epileptogenic process, and there are no generally accepted drugs recognized as anti-epileptogenic.

SUMMARY

Provided herein is a method for reducing the likelihood of epileptogenesis in a subject (e.g., a subject at risk thereof), comprising administering to said subject (e.g., a human subject) an effective amount a peptide of Formula I: X1-X2-X3-X4-X5 (SEQ ID NO: l)

or a pharmaceutically acceptable salt thereof, wherein

XI is V or R;

X2 is S, A, or H;

X3 is K or R;

X4 is K or R; and

X5 is R, L, or K.

Also provided is the use of a peptide of Formula I or a pharmaceutically acceptable salt thereof as taught herein for reducing the likelihood of epileptogenesis in a subject, or for the preparation of a medicament for reducing the likelihood of epileptogenesis in a subject.

In some embodiments, the peptide is N-terminal acetylated and/or C-terminal amidated. In some embodiments, the peptide of Formula I is: VSRKR (SEQ ID NO:2), VSKRR (SEQ ID NO:3), VSRRR (SEQ ID NO:4), VARKL (SEQ ID NO:5), RHKKL (SEQ ID NO:6), RARRL (SEQ ID NO:7), RSKKL (SEQ ID NO:8), RHKRR (SEQ ID NO:9), VARRL (SEQ ID NO: 10), VARRK (SEQ ID NO: 11), or RSKRR (SEQ ID NO: 12).

In some embodiments, the peptide of Formula I is VSKRR (SEQ ID NO:3). In some embodiments, the peptide of Formula I is VSRRR (SEQ ID NO:4).

In some embodiments, subjects at risk for epileptogenesis have brain injury, such as a traumatic brain injury, especially penetrating brain injury or depressed skull fracture, intraaxial or extraxial tumors or masses, stroke, parasitic lesions, or global hypoxia-ischemia. In some embodiments, subjects at risk for epileptogenesis have non-penetrating brain injury, brain tumor, intracranial hemorrhage, subarachnoid hemorrhage, prolonged febrile seizures, CNS or brain infection (meningitis or encephalitis), prior seizures or status epilepticus stroke, etc. In some embodiments, subjects at risk for epileptogenesis have T2 magnetic resonance imaging (MRI) signal changes (e.g., increased T2 signal) in temporal lobe structures such as the hippocampus.

In some embodiments, the method further includes concurrently or sequentially administering to said subject an anti-convulsant agent.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 presents the results of administration of CN-105 in preclinical model epileptogenesis. DETAILED DESCRIPTION

The disclosures of all patent references cited herein are hereby incorporated by reference to the extent they are consistent with the disclosure set forth herein. As used herein in the description of the invention and the appended claims, the singular forms "a," "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

"Epileptogenesis" as used herein refers to the pathological process of developing epilepsy. Epilepsy, as known in the art, is a chronic condition characterized by multiple spontaneous recurrent seizures. As further described below, risk factors for epileptogenesis include, but are not limited to, head injury (especially penetrating), induction of status epilepticus (e.g., as with febrile seizures), central nervous system (CNS) infection, tumor, mass lesion, hemorrhage, stroke, etc.

A "peptide" as used herein refers to a compound that comprises amino acid residues (and/or amino acid mimetic(s)), or a derivative thereof. Amino acids are well known in the art and include, for example, isoleucine, leucine, alanine, asparagine, glutamine, lysine, aspartic acid, glutamic acid, methionine, cysteine, phenylalanine, threonine, tryptophan, glycine, valine, proline, serine, tyrosine, arginine, histidine, norleucine, ornithine, taurine, selenocysteine, selenomethionine, lanthionine, 2-aminoisobutyric acid, dehydroalanine, hypusine, citrulline, 3- aminopropanoic acid, gamma-aminobutryic acid, nitroarginine, N-methylated leucine, homoarginine, dimethyl arginine, acetyl lysine, azalysine, pyrrolysine, and the like. An "amino acid side chain" refers to the various organic substituent groups that differentiate one amino acid from another. An amino acid having a hydrophobic side chain includes the non-limiting examples of alanine (A), isoleucine (I), leucine (L), methionine (M), phenylalanine (F), tryptophan (W), tyrosine (Y), and valine (V). An amino acid having a positively charged side chain, under typical physiological conditions, includes the non-limiting examples of arginine (R), histidine (H), and lysine (K). An amino acid having a negatively charged side chain, under typical physiological conditions, includes the non-limiting examples of aspartic acid (D) and glutamic acid (E). An amino acid having a polar uncharged side chain includes the non-limiting examples of serine (S), threonine (T), asparagine (N), and glutamine (Q). A "derivative" of an amino acid side chain refers to an amino acid side chain that has been modified structurally (e.g. , through chemical reaction to form a new species, covalent linkage to another molecule, and the like). Some embodiments provide for a peptide comprising modifications including, but not limited to, glycosylation, side chain oxidation, acetylation, amidation, or phosphorylation, as long as the modification does not destroy the biological activity of the peptides as herein described. For example, in some embodiments, a peptide may be modified by N-terminal acetylation and/or C-terminal amidation.

An "amino acid mimetic" as used herein is meant to encompass peptidomimetics, peptoids (poly-N-substituted glycines) and β-peptides (i.e., peptides that comprise one or more amino acids residues having the amino group attached at the β-carbon rather than the a-carbon). Suitably, the amino acid mimetic comprises an altered chemical structure that is designed to adjust molecular properties favorably (e.g., stability, activity, reduced immunogenic response, solubility, etc.). Typically, the altered chemical structure is thought to not occur in nature (e.g., incorporating modified backbones, non-natural amino acids, etc.). Thus, non-limiting examples of amino acid mimetic include D-peptides, retro-peptides, retro-inverso peptides, β-peptides, peptoids, and compounds that include one or more D-amino acids, poly-N-substituted glycine, or β-amino acid, or any combination thereof.

Typically, a peptide comprises a sequence of at least 3 amino acids (amino acid residues) and/or amino acid mimetics. Embodiments of the disclosure relate to small peptides of at least 3, 4, 5, 6, 7, 8, or 9 amino acid residues, mimetics, or combinations thereof. Some embodiments described herein provide for peptides of fewer than 9, 8, 7, 6, 5, or 4 amino acid residues and/or mimetics. Some embodiments relate to peptides that are 5 amino acids in length. The peptides described herein can be provided in a charged form, typically with a net positive charge, and can be generated and used as salts (e.g. , alkali metal salts, basic or acidic addition salts). The selection and formation of such salts are within the ability of one skilled in the art. See, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins, A Wolters Kluwer Company, Philadelphia, Pa (2005).

Embodiments of the disclosure provide synthetic peptides with ApoE mimetic activity. Though they may exhibit ApoE mimetic activity, the disclosed peptides do not share primary protein sequence identity with the native ApoE polypeptide. In other words, the disclosed peptide sequences do not appear in the primary amino acid sequence of an ApoE polypeptide, nor do they exhibit a-helical secondary structure analogous to the native ApoE receptor-binding domain. In an embodiment, the synthetic peptides are optionally isolated and/or purified to a single active species.

In an aspect of the disclosure, the peptide comprises Formula I:

X1-X2-X3-X4-X5 (SEQ ID NO:l)

or a salt thereof, wherein XI is selected from an amino acid having a hydrophobic side chain or an amino acid having a positively charged side chain; X2 is selected from an amino acid having a hydrophobic side chain, an amino acid having a positively charged side chain, or an amino acid having a polar uncharged side chain; X3 is selected from an amino acid having a positively charged side chain; X4 is selected from an amino acid having a positively charged side chain; and X5 is selected from an amino acid having a hydrophobic side chain or an amino acid having a positively charged side chain. Some embodiments provide a peptide wherein XI is V or R; X2 is S, A, or H; X3 is K or R; X4 is K or R; and X5 is R, L, or K.

Some embodiments of this aspect provide for a peptide of Formula II:

X1-X2-X3-X4-X5-X6-X7-X8-X9 (SEQ ID NO: 13);

wherein XI, X2, X3, X4, and X5 are as noted above, and each of X6, X7, X8, and X9 are independently selected from any amino acid, and are optionally absent. In such embodiments, the peptide of Formula II can include 5 amino acid residues, 6 amino acid residues, 7 amino acid residues, 8 amino acid residues, or 9 amino acid residues.

A number of non-limiting embodiments of peptides according to Formula I are disclosed in Table 1. In some embodiments the peptide can comprise VSRKR (SEQ ID NO:2), VSKRR (SEQ ID NO:3), VSRRR (SEQ ID NO:4), VARKL (SEQ ID NO:5), RHKKL (SEQ ID NO:6), RARRL (SEQ ID NO:7), RSKKL (SEQ ID NO:8), RHKRR (SEQ ID NO:9), VARRL (SEQ ID NO:10), VARRK (SEQ ID NO:l l), or RSKRR (SEQ ID NO: 12). In some embodiments, the peptide according to Formula I is VSRKR (SEQ ID NO:2), VSKRR (SEQ ID NO:3), VSRRR (SEQ ID NO:4), VARKL (SEQ ID NO:5), RHKKL (SEQ ID NO:6), RARRL (SEQ ID NO:7), RSKKL (SEQ ID NO:8), RHKRR (SEQ ID NO:9), VARRL (SEQ ID NO: 10), VARRK (SEQ ID NO: 11), or RSKRR (SEQ ID NO: 12).

In some embodiments of all the aspects described herein, the peptides consist essentially of the amino acid sequences and formulae disclosed herein. In some embodiments of all the aspects described herein, the peptides consist of the amino acid sequences and formulae disclosed herein. See also U.S. Patent Nos. 9,018,169 and 9,303,063 to Laskowitz et al.

Table 1.

Peptide Sequence

SEQ ID NO: 1 XI X2 X3 X4 X5

SEQ ID NO:2 V S R K R

SEQ ID NO:3 V S K R R

SEQ ID NO:4 V S R R R

SEQ ID NO:5 V A R K L

SEQ ID NO:6 R H K K L SEQ ID NO:7 R A R R L

SEQ ID NO:8 R S K K L

SEQ ID N0:9 R H K R R

SEQ ID NO: 10 V A R R L

SEQ ID NO: 11 V A R R K

SEQ ID NO: 12 R S K R R

In some embodiments, the peptides can exhibit at least one ApoE mimetic activity. In some embodiments, for example, the disclosed peptides can bind one or more physiological ApoE receptors such as, for example, cell-surface receptors expressed by glial cells, as well as receptors that function to suppress the neuronal cell death and calcium influx (excitotoxicity) associated with N-methyl-D-aspartate (NMDA) exposure; protect against LPS-induced production of TNF-a and IL-6 (e.g., in an in vivo sepsis model); prevent, treat, or slow inflammatory disorders such as atherosclerosis, arthritis, or inflammatory bowel disease; suppress glial or microglial activation; suppress macrophage activation; suppress lymphocyte activation; suppress inflammation; suppress CNS inflammation; treat neuropathy; and/or ameliorate neuronal injury in neurodegenerative disease (e.g. , mild cognitive impairment, dementia, Parkinson's disease, or Alzheimer's disease) and/or acute CNS trauma (e.g., traumatic brain injury).

The peptides can be produced using any means for making polypeptides known in the art, including, e.g., synthetic and recombinant methods. For example, in some embodiments the peptides can be synthesized using synthetic chemistry techniques such as solid-phase synthesis, Merrifield-type solid-phase synthesis, t-Boc solid-phase synthesis, Fmoc solid-phase synthesis, BOP solid-phase synthesis, and solution-phase synthesis. See, for example, Stewart and Young, Solid Phase Peptide Synthesis, 2nd ed., (1984) Pierce Chem. Co., Rockford 111.; The Peptides: Analysis, Synthesis, Biology, Gross and Meienhofer, Eds., vols. 1-2 (1980) Academic Press, New York; Bodansky, Principles of Peptide Synthesis, (1984) Springer-Verlag, Berlin. In other embodiments, the peptides can be produced, for example, by expressing the peptide from a nucleic acid encoding the peptide in a cell or in a cell-free system according to recombinant techniques familiar to those of skill in the art. See, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel et al., Current Protocols in Molecular Biology, (2002) John Wiley & Sons, Somerset, NJ; each of which is hereby incorporated by reference in its entirety. The peptides can incorporate any of the various modifications and protective groups described herein or otherwise known to those of skill in the art, such as, for example, those described in McOmie, Protective Groups in Organic Chemistry, (1973) Plenum Press, New York.

Particular embodiments of peptide active agents as described above (shown in the form of their chemical structures), and for use in the methods and compositions described herein include, but are not limited to, the following, along with pharmaceutically acceptable salts thereof:



 The present disclosure provides a method of reducing the likelihood of developing epilepsy (i.e., reducing the risk of epileptogenesis) in a subject in need thereof (i.e., a subject at risk of epileptogenesis), the method comprising administering to the subject an effective amount of a peptide of Formula I or Formula II, or a composition or formulation comprising an effective amount of a peptide of Formula I or Formula II.

Subjects "at risk for epileptogenesis" include, but are not limited to, those with brain injury, including traumatic brain injury, especially penetrating brain injury or depressed skull fracture, intraaxial or extraxial tumors or masses, stroke, parasitic lesions, and global hypoxia- ischemia. Other risk factors may include, but are not limited to, non-penetrating brain injury, brain tumor, intracranial hemorrhage, subarachnoid hemorrhage, prolonged febrile seizures, CNS or brain infection (meningitis or encephalitis), patients with prior seizures or status epilepticus stroke, etc.

In some embodiments, subjects at risk for epileptogenesis have T2 magnetic resonance imaging (MRI) signal changes (e.g., increased T2 signal) in temporal lobe structures such as the hippocampus. See, e.g., Shinnar et al., "MRI abnormalities following febrile status epipelpticus in children," Neurology 79(9):871-877 (2012).

Subjects described herein include mammalian subjects, including both human subjects and non-human (animal) subjects, though in some embodiments human subjects are preferred. Subjects may also include non-human primates, rodents such as mice and rats, dogs, cats, rabbits, goats, horses, pigs, cattle, etc., for research and/or veterinary purposes.

Subjects include both male and female subjects and subjects of all ages including infant, juvenile, adolescent and adult subjects. In some embodiments, subjects are infant or juvenile subjects, e.g., those with febrile seizures or CNS infection such as meningitis, which increases their risk of epileptogenesis.

In some embodiments, the subject may also be concurrently treated with an anticonvulsant agent as a prophylactic treatment (i.e., a prophylactic antiepileptic drug (AED)), for example, to treat early seizures (i.e., seizures occurring within seven days of the CNS injury or insult). The term "concurrently administered" as used herein means that two compounds are administered sufficiently close in time to achieve a combined effect. Concurrent administration may thus be carried out by sequential administration or simultaneous administration (e.g., simultaneous administration in a common, or the same, carrier).

Numerous anti-convulsant agents are known and include, but are not limited to, aldehydes such as paraldehyde; aromatic allylic alcohols such as stiripentol; barbiturates such as phenobarbital, methylphenobarbital, barbexaclone; benzodiazepines such as clobazam, clonazepam, clorazepate, diazepam, midazolam, Iorazepam, nitrazepam, temazepam, nimetazepam; bromides such as potassium bromide; carbamates such as carbamazepine (Tegretol), oxcarbazepine and eslicarbazepine acetate; fatty acids such as valproate (valproic acid (DEPAKOTE®), sodium valproate, divalproex sodium), vigabatrin, progabide, tiagabine; fructose derivatives such as topiramate; GABA analogs such as gabapentin, pregabalin; hydantoins such as phenytoin (DILANTIN®), mephenytoin (Mesantoin), ethotoin, fosphenytoin; oxazolidinediones such as paramethadione, trimethadione (Tridione), ethadione; propionates such as beclamide; pyrimidinediones such as primidone (Mysoline); pyrrolidines such as brivaracetam, levetiracetam, seletracetam; succinimides such as ethosuximide (Zarontin), phensuximide, mesuximide; sulfonamides such as acetazolamide, sultiame, methazolamide, zonisamide; triazines such as lamotrigine; ureas such as pheneturide, phenacemide; valproylamides such as valpromide, valnoctamide; perampanel, etc., as well as pharmaceutically acceptable salts and/or prodrugs thereof.

In some embodiments, the anti-convulsant is levetiracetam, ethosuximide, phenytoin, phenobarbital, carbamazepine, valproate, lacosamide, clobazam (Onfi), brivaracetam (Briviact), etc.

In some embodiments, the disclosed peptides and compositions may be administered by any suitable route of administration, including, but not limited to, injection (subcutaneous, intraperitoneal, intravenous, intrathecal, intramuscular, intracerebroventricular, and spinal injection), intranasal, oral, transdermal, parenteral, inhalation, nasopharyngeal or transmucosal absorption. In some embodiments, the peptides may be delivered by injection, inhalation, transdermal, intravenous, intranasal, intracranial, and/or intrathecal administration. In some embodiments, the peptides may be delivered intravenously (e.g., provided in normal saline). Administration encompasses the providing at least one peptide as described herein (e.g., of Formula I, Formula II, and/or SEQ ID NOs: l-12) formulated as a pharmaceutical composition. Administration of an active agent (e.g., compound, peptide, etc.) directly to the brain is known in the art. Intrathecal injection delivers agents directly to the brain ventricles and the spinal fluid. Surgically-implantable infusion pumps are available to provide sustained administration of agents directly into the spinal fluid. Spinal injection involves lumbar puncture with injection of a pharmaceutical compound into the cerebrospinal fluid. Administration also includes targeted delivery wherein peptide according to the disclosure is active only in a targeted region of the body (for example, in brain tissue), as well as sustained release formulations in which the peptide is released over a period of time in a controlled manner. Sustained release formulations and methods for targeted delivery are known in the art and include, for example, use of liposomes, drug loaded biodegradable microspheres, drug-polymer conjugates, drug-specific binding agent conjugates and the like.

Pharmaceutically acceptable carriers are well known to those of skill in the art and may include, but are not limited to, sterile pyrogen-free water, sterile pyrogen-free physiological saline solution, etc. Pharmaceutically acceptable carriers may also include chitosan nanoparticles or other related enteric polymer formulations.

Particular pharmaceutical formulations and therapeutically effective amounts and dosing regimens may be determined by one of skill in the art based upon the disclosures herein and taking into consideration, for example, a subject's age, weight, sex, ethnicity, organ (e.g., liver and kidney) function, the extent of desired treatment, the stage and severity or risk of the disease or disorder and associated symptoms, and the tolerance of the subject. In some embodiments, effective amounts of the peptides disclosed herein can be about 0.1 or 0.5 to 5, 8 or 10 mg/kg, for example, about 1 mg/kg. In some embodiments, dosage regimens may include, but are not limited to, dosages once every 4, 5, 6, 7 or 8 hours, for 60, 72, or 90 hours after insult or injury. For example, the peptides may be administered once every 6 hours for 72 hours (13 doses total).

In aspects described herein that relate to compositions, including pharmaceutical compositions and formulations, some embodiments provide a composition that comprises at least one peptide according to SEQ ID NO: l (e.g., SEQ ID NOs: l-12), or a salt thereof (e.g., an acetate salt) in combination with an acceptable carrier.

The present invention is explained in greater detail in the following non-limiting examples.

EXAMPLES

Pilocarpine Induction Repeated Injection Model of Epileptogenesis. This model of epilepsy induction in mice involves the use of repeated pilocarpine injections model modified from Groticke (Grctticke et al. / Experimental Neurology 207 (2007) 329-349) briefly described here. In this model, there is a latent period that follows the acute seizures induced by the pilocarpine. About 10-14 days later, the animal begins to have spontaneous, recurrent, clinically apparent seizures (epilepsy), as a model of epileptogenesis.

C57B6 mice (Charles Rivers) are injected with 2mg/kg of methylscopolamine (Sigma S8502) 15 minutes before a bolus dose of 350-450 mg/kg pilocarpine, with repeated injections 100 mg/kg every 20 minutes until animal has observable seizures. After animals are in status epilepticus (having seizures) for 45 minutes after the onset of observable seizures, animals are then randomized and administered either 0.9 % saline or (0.1-1.0 mg /kg) of CN-105 (acetyl - Val-Ser-Arg-Arg-Arg-NH₂) + saline. At 50 minutes after the onset of observable seizures animals are injected with 10 mg/kg of diazepam, and a second injection of diazepam is given at 80 minutes after the onset of observable seizures to stop status. A second injection of 0.9 % saline or CN-105 is given 6 hours after start of seizures, and/or daily doses Q12 or Q24 for up 7 days. Lead placement is done 7-14 days after Day one inductions, followed by video Electroencephalography (EEG) monitoring up to 30 days after lead placement. EEG reader is blinded to treatment; all seizures are confirmed by video review and/or a second experience reader. Data is presented as a percent; total number of mice that are epileptic divided by total number of mice in each treatment group. The number of seizers, seizures per mouse, seizures per day, seizure length and type of seizures can all be extracted from the EEG review.

Results are shown in FIG. 1. The data demonstrate that administering CN-105 reduces the development of epilepsy as a disease modifying agent. This outcome is in contrast to traditional anti-epileptic drugs (AEDs), which symptomatically suppress seizures but do not change the underlying epileptogenesis.

The administration of CN-105 in the first few hours after status has no impact on the ongoing seizures with the first injection, and there are no ongoing seizures at the 6 hour second injection. Thus, the administration of CN-105 reduces the likelihood of developing the subsequent, late seizures at 10-14 days.

This effect is in contrast to the use of anticonvulsants, which can reduce early seizures, but do not stop the development of later seizures. In the case of CN-105, early seizures were not stopped, but the development of later seizures (i.e., epileptogenesis) was reduced.

Peptides similar to CN-105 are also predicted to be effective for reducing the likelihood of epileptogenesis. See also U.S. Patent No. 9,018,169 to Laskowitz et al, which demonstrates anti-inflammatory activity for various peptides of Formula I.

The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

What is Claimed is:

1. A peptide of Formula I:

X1-X2-X3-X4-X5 (SEQ ID NO:l)

or a pharmaceutically acceptable salt thereof, wherein

XI is V or R;

X2 is S, A, or H;

X3 is K or R;

X4 is K or R; and

X5 is R, L, or K,

optionally wherein said peptide is N-terminal acetylated and/or C-terminal amidated,

for use in reducing the likelihood of epileptogenesis in a subject.

2. The peptide of claim 1 for use of claim 1 , wherein the peptide of Formula I is: VSRKR (SEQ ID NO.-2), VSKRR (SEQ ID NO:3), VSRRR (SEQ ID NO:4), VARKL (SEQ ID NO:5), RHKKL (SEQ ID NO:6), RARRL (SEQ ID NO:7), RSKKL (SEQ ID NO:8), RHKRR (SEQ ID NO:9), VARRL (SEQ ID NO: 10), VARRK (SEQ ID NO:l 1), or RSKRR (SEQ ID NO: 12), or a pharmaceutically acceptable salt thereof.

3. The peptide of claim 1 for use of claim 1, wherein the peptide of Formula I is selected from the group consisting of:

15



and pharmaceutically acceptable salts thereof.

The peptide of claim 1 for use of claim 1, wherein the peptide of Formula I is:

or a pharmaceutically acceptable salt thereof (e.g., acetate salt).

5. The peptide of any one of claims 1-4 for use of any one of claims 1-4, wherein said peptide is provided in a pharmaceutically acceptable carrier.

6. The peptide of claim 5 for use of claim 5, wherein said pharmaceutically acceptable carrier is suitable for administering said peptide by injection or infusion (e.g. , sterile, pyrogen- free physiological saline solution).

7. The peptide of any one of claims 1 -6 for use of any one of claims 1-6, wherein said subject has traumatic brain injury.

8. The peptide of claim 7 for use of claim 7, wherein said traumatic brain injury comprises a penetrating brain injury.

9. The peptide of claim 7 for use of claim 7, wherein said traumatic brain injury comprises a depressed skull fracture.

10. The peptide of any one of claims 1-9 for use of any one of claims 1-9, wherein said subject has a family history of epilepsy.

11. The peptide of any one of claims 1-10 for use of any one of claims 1-10, wherein said subject has a history of prolonged febrile seizures.

12. The peptide of any one of claims 1-1 1 for use of any one of claims 1-11, wherein said subject has T2 MRI signal changes in the hippocampus.

13. The peptide of any one of claims 1-12 for use of any one of claims 1-12, wherein said subject has or has had a central nervous system infection (e.g., meningitis).

14. The peptide of any one of claims 1-13 for use of any one of claims 1-13, wherein said subject is an adult subject.

15. The peptide of any one of claims 1-13 for use of any one of claims 1-13, wherein said subject is an infant or juvenile subject.

16. The peptide of any one of claims 1-15 for use of any one of claims 1-15, wherein said subject is a human subject.

17. The peptide of any one of claims 1 -16 for use of any one of claims 1-16, wherein said peptide is for injection, inhalation, transdermal, intravenous, intranasal, intracranial, and/or intrathecal administration.

18. The peptide of any one of claims 1-16 for use of any one of claims 1-16, wherein said peptide is for intravenous administration.

19. The peptide of any one of claims 1-18 for use of any one of claims 1-18, wherein the peptide is administered at a dosage of from 0.1 to 10 mg/kg once every 4-8 hours, for 60-90 hours total.

20. The peptide of claim 19 for use of claim 19, wherein the peptide is administered at a dosage of about 1 mg/kg once every 6 hours for 72 hours total (13 doses total).

21. The peptide of any one of claims 1-20 for use of any one of claims 1-20, wherein said peptide is administered to said subject concurrently or sequentially with an anti-convulsant agent.

22. A method for reducing the likelihood of epileptogenesis in a subject at risk thereof, comprising administering to said subject an effective amount a peptide of Formula I:

X1-X2-X3-X4-X5 (SEQ ID NO:l)

or a pharmaceutically acceptable salt thereof, wherein

XI is V or R;

X2 is S, A, or H;

X3 is K or R;

X4 is K or R; and

X5 is R, L, or K,

optionally wherein said peptide is N-terminal acetylated and/or C-terminal amidated.

23. The use of a peptide of Formula I or pharmaceutically acceptable salt thereof for the preparation of a medicament for reducing the likelihood of epileptogenesis in a subject.