Methods of treating and inhibiting progression of epileptic seizures

Abstract

Methods of treating epileptic seizures by extracranial administration of a clostridium derivative and methods of inhibiting the development of focal epileptic seizures to systemic epileptic seizures are described.

Description

[0001] This application is a divisional application of the Chinese invention patent application with application number 202080057952.0 filed on July 6, 2020 and invention name “Method for treating and inhibiting the development of epileptic seizures”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This patent application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application Serial No. 62 / 870,872, filed on July 5, 2019, which is incorporated herein by reference in its entirety. Technical Field

[0004] The present disclosure relates to methods of treating epileptic seizures and methods of inhibiting the progression of focal epileptic seizures to generalized epileptic seizures by extracranial administration of Clostridial derivatives. Background Art

[0005] Epileptic seizures are usually associated with abnormal paroxysmal brain electrical activity. The clinical manifestations of epileptic seizures can include changes in consciousness and movement, sensation, autonomy or mental events perceived by the patient or observer. Typically, epileptic seizures cause temporary loss of consciousness, convulsions, changes in muscle tone, abnormal movements and / or confusion. Epileptic seizures can have many causes, such as drugs, high fever, craniocerebral injury, genetic etiology, infectious diseases, metabolic disorders, stroke and other diseases. Recurrent unprovoked epileptic seizures define a neurological syndrome known as epilepsy. Epileptic seizures can be diagnosed by observation or by recording brain electrical activity, such as electroencephalogram (EEG).

[0006] Epileptic seizures are classified into three categories based on the symptoms perceived by the patient or observer and / or the pattern of electrical activity accompanying the seizure. Broadly speaking, seizures can be classified as focal, generalized, and unclassified.

[0007] Focal (also called "partial") seizures are those that occur in only a portion of one cerebral hemisphere, as indicated by initial symptomatology or EEG findings. Of note, focal seizures typically present as asymptomatic abnormal electrical activity in only a small amount of cortex in one hemisphere. Depending on their very localized onset, focal seizures spread to adjacent cortical areas within a hemisphere, at which point they become perceptible to the patient or observer.

[0008] Generalized seizures are those in which initial semiology or EEG findings indicate minimal involvement of more than one cerebral hemisphere and can be subdivided into convulsive and nonconvulsive seizures.

[0009] Another subclassification of generalized seizures is primary generalized seizures that arise from both hemispheres simultaneously, and secondary generalized seizures that arise focally in one hemisphere and then spread to involve the other. The third category, unclassified seizures, includes all seizures that are difficult to classify due to incomplete data.

[0010] Epilepsy affects 1% of the world's population. In individuals with epilepsy who experience localized (focal or partial) seizures, consciousness may be disturbed and varying degrees of memory loss may occur, but seizures do not lead to loss of consciousness. However, focal seizures can develop into secondary generalized seizures, in which case individuals lose consciousness. Clinically, generalized and bilateral seizures are considered to be more dangerous because they are more likely to cause damage and post-epileptic complications. Epidemiologically, nearly half of all adults diagnosed with epilepsy experience at least one generalized or bilateral seizure, and about 25% of epilepsy patients often experience generalized or bilateral seizures. Current treatments for epilepsy and other epileptic seizures include antiepileptic drugs (AEDs). However, standard care drug therapy is not completely effective for many patients and can cause undesirable side effects. SUMMARY OF THE INVENTION

[0012] In one aspect, a method of treating seizures associated with epilepsy is provided. The method comprises administering to a subject suffering from epilepsy a Clostridium derivative in an amount effective to treat the seizures, wherein the Clostridium derivative is administered extracranially to at least one administration site.

[0013] In one embodiment, a method of treating epilepsy is provided. The method comprises administering to a subject suffering from epilepsy a therapeutically effective amount of a Clostridial derivative, wherein the Clostridial derivative is administered extracranially to at least one administration site.

[0014] In one embodiment, a method for treating symptoms of premonitory epileptic seizures (or premonitory epilepsy) is provided. The method comprises administering a therapeutically effective amount of a Clostridium derivative to a subject suffering from symptoms of premonitory epileptic seizures (or premonitory epilepsy), wherein the Clostridium derivative is administered extracranially to at least one administration site.

[0015] In one embodiment, the Clostridium derivative is administered by an injection route selected from the group consisting of extramuscular injection, subcutaneous injection, subdermal injection, or intradermal injection.

[0016] In one embodiment, the Clostridium derivative is administered by injection into the scalp.

[0017] In one embodiment, the Clostridium derivative is administered to a suture in the cervical region of the scalp. In one embodiment, the Clostridium derivative is administered to a suture in the cervical region of the scalp. In one embodiment, the Clostridium derivative is administered to or near the lambdoid suture, the parieto-mammary suture, the occipital suture, or a combination thereof. In an alternative embodiment, the Clostridium derivative is administered to a muscle in the cervical region of the scalp. In another embodiment, the Clostridium derivative is administered to a cervical nerve in the cervical region. In another alternative embodiment, the Clostridium derivative is administered to the trigeminal nerve in the cervical region.

[0018] In another embodiment, the Clostridium derivative is administered to at least one suture on the patient's skull. In one embodiment, the Clostridium derivative is administered to a cervical nerve. In another embodiment, the Clostridium derivative is administered to a trigeminal nerve. In another embodiment, the Clostridium derivative is administered to a muscle on the patient's skull.

[0019] In another embodiment, the method comprises administering the Clostridium derivative to more than one administration site. In an exemplary embodiment, the number of administration sites is between 2-8.

[0020] In one embodiment, the at least one administration site is generally along the lambdoid suture.

[0021] In another embodiment, the at least one administration site is approximately at the junction of the lambdoid and sagittal sutures.

[0022] In another embodiment, the at least one administration site is near and / or generally along any suture of the skull, selected from, for example, the frontal suture, squamous suture, coronal suture, lambdoid suture, occipitomastoid suture, parietomastoid suture, and sagittal suture.

[0023] In one embodiment, the at least one administration site is in the occipital region innervated by the cervical nerves. In a specific embodiment, onabotulinumtoxinA is administered by injection to the occipital region innervated by the cervical nerves.

[0024] In one embodiment, the at least one administration site is any structure or tissue proximal to the trigeminal nerve or cervical nerve, including, for example, fascia, subfascia, aponeurosis, epineurium, or general connective tissue, muscle, nerve, and sutures.

[0025] In other embodiments, the amount of the Clostridial derivative administered is evenly divided into portions for administration to each administration site. In other embodiments, a portion of the amount of the Clostridial derivative is administered to each administration site, wherein the portion administered to each administration site is unequal.

[0026] In other embodiments, the Clostridium derivative is a botulinum neurotoxin. In one embodiment, the botulinum neurotoxin is botulinum toxin type A, botulinum toxin type B, botulinum toxin type C, botulinum toxin type D, or botulinum toxin type E.

[0027] In one embodiment, the botulinum toxin is a botulinum toxin type A. In some embodiments, the botulinum toxin type A is selected from onabotulinumtoxinA, incobotulinumtoxinA, abobutulinumtoxinA, daxibotulinumtoxinA, prabotulinumtoxinA, and combinations thereof. In some embodiments, the amount of the botulinum neurotoxin type A administered is between about 1 unit and 3000 units. In some embodiments, the amount of the botulinum neurotoxin type A administered is between 2 units and 200 units. In some other embodiments, the amount of the botulinum neurotoxin type A administered is between 200 units and 500 units. In other embodiments, the amount of the botulinum neurotoxin administered is greater than 500 units.

[0028] In one embodiment, the botulinum toxin comprises a botulinum toxin complex. In another embodiment, the botulinum toxin comprises a pure neurotoxin.

[0029] In one embodiment, the botulinum toxin is onabotulinumtoxin A and is administered in an amount of about 2-200 units. In one embodiment, the administered amount is evenly divided into about 2-10 administration sites.

[0030] In other embodiments, the seizure is selected from a focal seizure, a myoclonic seizure, and a generalized tonic-clonic seizure.

[0031] In another embodiment, the seizure is associated with an epilepsy disorder selected from the group consisting of epilepsy, juvenile myoclonic epilepsy, idiopathic generalized epilepsy, and Lennox-Gastaut syndrome.

[0032] In other embodiments, the seizure is a generalized tonic-clonic seizure associated with Lennox-Gastaut syndrome.

[0033] In one embodiment, the step of administering the Clostridium derivative is an adjunctive therapy for treating epileptic seizures by oral therapy, subcutaneous, rectal or intravenous administration.

[0034] In another embodiment, the step of administering the Clostridium derivative is an adjunct therapy for treating epileptic seizures by electrically stimulating the vagus nerve, or the facial nerve, or the thalamus, or the cerebral cortex. On the other hand, a method for inhibiting the development of focal epileptic seizures into bilateral epileptic seizures is provided. The method comprises administering a Clostridium derivative extracranially to a subject suffering from focal epileptic seizures in a therapeutically effective amount to inhibit the spread of abnormal brain electrical activity to nearby brain areas, thereby preventing the spread and / or generalization of focal epileptic seizures.

[0035] In another aspect, a method for preventing focal asymptomatic epileptic seizures from developing into focal symptomatic epileptic seizures is provided. The method comprises administering a Clostridium derivative extracranially to a subject suffering from focal epileptic seizures in a therapeutically effective amount to inhibit the spread of abnormal brain electrical activity to nearby brain regions, thereby preventing focal asymptomatic epileptic seizures from developing into focal symptomatic epileptic seizures.

[0036] In another aspect, a method of treating seizures associated with epilepsy is provided, the method comprising extracranially administering a Clostridium derivative to a subject having seizures in an amount therapeutically effective to treat the seizures.

[0037] In another aspect, a method of treating seizures in a subject suffering from epilepsy is provided, comprising extracranially administering to the subject a therapeutically effective amount of a Clostridium derivative to treat the seizures.

[0038] In another aspect, a method of controlling focal seizures comprises extracranially administering to a subject a Clostridium derivative in an amount effective to control focal seizures to a therapeutically effective amount.

[0039] In one embodiment, the Clostridium derivative is administered to the scalp.

[0040] In some embodiments, the methods according to the present disclosure further comprise administering to the subject an adjunctive therapy for epileptic seizures. In some embodiments, the adjunctive therapy comprises one or more anti-epileptic drugs.

[0041] In another aspect, a method of treating episodic headache is provided, the method comprising administering to a subject extracranially a Clostridial derivative in an amount effective for treating episodic headache. In one embodiment, the Clostridial derivative is administered before the onset of epilepsy (pre-ictal). In another embodiment, the Clostridial derivative is administered after the onset of epilepsy (post-ictal). BRIEF DESCRIPTION OF THE DRAWINGS

[0043] The following figures are provided to illustrate various aspects and features of embodiments of the present method.

[0044] Figure 1Ais an illustrative top view of an experimental animal showing certain cranial sutures and administration sites for delivering a Clostridium derivative in one embodiment;

[0045] Figure 1B is a top view of an illustrative cortical anatomy showing the location of picrotoxin administration for inducing epileptic seizures and the location of glass pipette electrodes for recording electrocorticograms in different cortical areas;

[0046] Figure 2A-2B The occipital cortex of experimental animals was examined before and after local application of picrotoxin to induce focal epileptic seizures. Figure 2A ) and parietal cortex ( Figure 2B ) of the cerebral cortical electrogram trace, the time of picrotoxin administration was determined by Figure 2A The dotted line in the

[0047] Figure 3A-3B The occipital cortex of experimental animals ( Figure 3A ) and parietal cortex ( Figure 3B ) of animals subjected to picrotoxin-induced epileptic seizures ( Figure 3A pre-treated with an exemplary Clostridium derivative (onabotulinumtoxinA) prior to administration at the time indicated by the dashed line in FIG.

[0048] Figures 4A-4B It is a human skull ( Figure 4A ) and rat skull ( Figure 4B ) showing the absence of the trigeminal nerve in the cervical region to which the Clostridium derivative was administered; and

[0049] Figure 5A , 5B and 5C is a schematic diagram of a human skull showing the various sutures.

[0050] Details

[0051] definition

[0052] The following definitions apply to this article:

[0053] As used herein, "about" or "approximately" means within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined (i.e., the limitations of the measurement system). For example, according to the practice in the art, "about" may mean within 1 or more than 1 standard deviation. Where specific values ​​are described in the application and claims, the term "about" means within an acceptable error range for the specific value unless otherwise indicated.

[0054] "Administration" or "to administer" refers to the step of giving (ie, administering) a pharmaceutical composition to a subject, or alternatively, the step of a subject receiving a pharmaceutical composition.

[0055] "Remission" refers to a decrease in the occurrence of epileptic seizures associated with a condition or disorder. Thus, remission includes some reduction, significant reduction, nearly complete reduction, and complete reduction.

[0056] "Biological activity" describes the beneficial or adverse effects of a drug on living substances. When the drug is a complex chemical mixture, this activity is exerted by the active ingredients of the substance but can be modified by other ingredients. Biological activity can be measured by LD in vivo 50 or ED 50 Assays may be assessed as efficacy or toxicity by in vitro assays, for example, cell-based potency assays as described in US20100203559 and US20100233802.

[0057] "Botulinum toxin" refers to a neurotoxin produced by Clostridium botulinum, as well as a botulinum toxin produced recombinantly from a non-Clostridium species, a fragment, variant or chimera thereof. As used herein, the term "botulinum toxin" includes serotype A botulinum toxin (BoNT / A), serotype B botulinum toxin (BoNT / B), serotype C botulinum toxin (BoNT / C), serotype D botulinum toxin (BoNT / D), serotype E botulinum toxin (BoNT / E), serotype F botulinum toxin (BoNT / F), serotype G botulinum toxin (BoNT / G), serotype H botulinum toxin (BoNT / H), serotype X botulinum toxin (BoNT / X), serotype En botulinum toxin (BoNT / En) and chimeric botulinum toxins and / or subtypes thereof and any other type of subtypes thereof, or in each case any re-engineered protein, analog, derivative, homolog, portion, subportion, variant or deformation of any of the foregoing. As used herein, "botulinum toxin" also includes "modified botulinum toxins." Other "botulinum toxins" as used herein also include botulinum toxin complexes (e.g., 300, 600, and 900 kDa complexes), and the neurotoxic component of botulinum toxin that is not bound to a complex protein (150 kDa).

[0058] "Clostridial derivative" refers to a molecule containing any portion of a Clostridial toxin as defined herein. As used herein, the term "Clostridial derivative" includes natural or recombinant neurotoxins, recombinant modified toxins, fragments, chimeras and variants thereof, targeted vesicle exocytosis modulators (TEMs) or combinations thereof.

[0059] "Clostridial toxin" refers to any toxin produced by a Clostridial toxin strain that can execute a whole cellular mechanism whereby the Clostridial toxin intoxicates a cell and includes binding of a Clostridial toxin to a low-affinity or high-affinity Clostridial toxin receptor, internalization of the toxin / receptor complex, translocation of the Clostridial toxin light chain into the cytoplasm, and enzymatic modification of Clostridial toxin substrates. Non-limiting examples of Clostridial toxins include botulinum toxins such as BoNT / A, BoNT / B, BoNT / C1, BoNT / D, BoNT / E, BoNT / F, BoNT / G, BoNT / H, BoNT / X, BoNT / En, chimeric botulinum toxins, tetanus toxin (TeNT), Baratii toxin (BaNT), and Butyricum toxin (BuNT). BoNT / C2 cytotoxin and BoNT / C3 cytotoxin, which are not neurotoxins, are excluded from the term "Clostridial toxin". Clostridial toxins disclosed herein include, but are not limited to, naturally occurring Clostridial toxin variants, such as Clostridial toxin isoforms and Clostridial toxin subtypes; non-naturally occurring Clostridial toxin variants, such as conservative Clostridial toxin variants, non-conservative Clostridial toxin variants, Clostridial toxin chimeric variants, and active Clostridial toxin fragments thereof, or any combination thereof. Clostridial toxins disclosed herein also include Clostridial toxin complexes and neurotoxic components of Clostridial toxins that are not bound to non-toxin associated proteins (NAPs). As used herein, the term "Clostridial toxin complex" refers to a complex comprising a neurotoxic component of a Clostridial toxin and a non-toxin associated protein (NAP), such as a botulinum toxin complex, a tetanus toxin complex, a Baratii toxin complex, and a butyric acid toxin complex. Non-limiting examples of Clostridial toxin complexes include those produced by Clostridium botulinum, such as a 900-kDa BoNT / A complex, a 500-kDa BoNT / A complex, a 300-kDa BoNT / A complex, a 500-kDa BoNT / B complex, a 500-kDa BoNT / C1 complex, a 500-kDa BoNT / D complex, a 300-kDa BoNT / D complex, a 300-kDa BoNT / E complex, and a 300-kDa BoNT / F complex.

[0060] A "Clostridial toxin active ingredient" refers to a molecule comprising any portion of a Clostridial toxin that exerts an effect when or after administration to a subject or patient. As used herein, the term "Clostridial toxin active ingredient" includes a Clostridial toxin complex comprising an approximately 150-kDa Clostridial toxin and other proteins collectively referred to as non-toxin associated proteins (NAPs), an approximately 150-kDa Clostridial toxin alone, or a modified Clostridial toxin, such as a retargeted Clostridial toxin.

[0061] "Effective amount" as applied to a biologically active ingredient refers to an amount of the ingredient that is generally sufficient to achieve the desired change in a subject. For example, when the desired effect is to relieve epileptic seizures or inhibit the progression of epileptic seizures, an effective amount of the ingredient is an amount that at least significantly reduces the frequency of epileptic seizures or the likelihood of focal epileptic seizures progressing to generalized epileptic seizures without causing significant toxicity. The effective amount of a Clostridium derivative can be expressed in units of potency or toxicity (e.g., LD 50 or ED 50 ) or mass units (e.g. ng or mg).

[0062] "Extracranial administration" refers to administration to a site outside the skull. In some embodiments, extracranial administration refers to administration to any suture, or combination of sutures, on a patient's skull.

[0063] "Topical administration" refers to administering a pharmaceutical agent at or near a site on or in an animal's body where its biological effect is desired, such as by, for example, intramuscular or intradermal or subdermal injection or topical administration. Topical administration does not include systemic routes of administration, such as intravenous or oral administration. Topical administration is a type of local administration in which the agent is applied to the patient's skin.

[0064] A "modified botulinum toxin" refers to a botulinum toxin in which at least one of its amino acids has been deleted, modified, or substituted compared to a native botulinum toxin. In addition, a modified botulinum toxin can be a recombinantly prepared neurotoxin, or a derivative or fragment of a recombinantly prepared neurotoxin. The modified botulinum toxin retains at least one biological activity of the native botulinum toxin, such as the ability to bind to a botulinum toxin receptor, or the ability to inhibit the release of a neurotransmitter from a neuron. An example of a modified botulinum toxin is a botulinum toxin having a light chain from one botulinum toxin serotype (such as serotype A) and a heavy chain from a different botulinum toxin serotype (such as serotype B). Another example of a modified botulinum toxin is a botulinum toxin conjugated to a neurotransmitter (such as substance P).

[0065] "Mutation" refers to a structural modification of a naturally occurring protein or nucleic acid sequence. For example, in the case of a nucleic acid mutation, the mutation may be a deletion, addition or replacement of one or more nucleotides in a DNA sequence. In the case of a protein sequence mutation, the mutation may be a deletion, addition or replacement of one or more amino acids in a protein sequence. For example, a specific amino acid that makes up a protein sequence may replace another amino acid, for example, an amino acid selected from the following: alanine, asparagine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, tyrosine or any other naturally occurring or non-naturally occurring amino acid or chemically modified amino acid. A mutation in a protein sequence may be the result of a mutation in a DNA sequence when the DNA sequence is transcribed and the resulting mRNA is translated, resulting in a mutated protein sequence. A mutation in a protein sequence may also be produced by fusing a peptide sequence containing the desired mutation to the desired protein sequence.

[0066] "Patient" refers to a human or non-human subject receiving medical or veterinary care. Thus, the compositions disclosed herein can be used to treat any animal, such as a mammal, etc.

[0067] "Administering peripherally" or "peripheral administration" refers to subdermal, intradermal, transdermal, subcutaneous administration or intramuscular administration.

[0068] A "pharmaceutical composition" refers to a composition comprising an active pharmaceutical ingredient, such as a Clostridial toxin (e.g., a botulinum toxin) and at least one additional ingredient (e.g., a stabilizer or excipient, etc.). Thus, a pharmaceutical composition is a formulation suitable for diagnostic or therapeutic administration to a subject (e.g., a human patient). A pharmaceutical composition can be, for example, in a lyophilized or vacuum-dried state, a solution formed after reconstitution of a lyophilized or vacuum-dried pharmaceutical composition, or a solution or solid that does not require reconstitution.

[0069] "TEMs", short for Targeted Exocytosis Modulators, are retargeted endopeptidases that direct the catalytic activity of the light chain to specific types of neuronal cells or target cells unaffected by botulinum toxin, which expands the beneficial clinical effects of inhibiting exocytosis in several human diseases.

[0070] "Treating" or "treatment" refers to temporarily or permanently alleviating (or eliminating) at least one symptom. More specifically, "treatment" of epileptic seizures, epilepsy, or symptoms thereof as used herein includes one or more of the following: reducing (or preventing) the frequency of epileptic seizures, all symptoms or symptoms thereof or reducing the duration of epileptic seizures, all symptoms or symptoms thereof, improving the severity of epileptic seizures, all symptoms or symptoms thereof, treating epileptic seizures, all symptoms or symptoms thereof, curing epileptic seizures, all symptoms or symptoms thereof, achieving a reduction in the number or severity of epileptic seizures or symptoms thereof, and improving the quality of life of patients experiencing epileptic seizures.

[0071] "Variant" refers to a clostridial neurotoxin, such as a wild-type botulinum toxin serotype A, B, C, D, E, F, G, H, X, or a chimeric botulinum toxin, which has been modified relative to the wild-type botulinum toxin by substitution, modification, addition or deletion of at least one amino acid, which is recognized by a target cell, internalized by a target cell and catalyzes the cleavage of a SNARE (SNAP (soluble NSF attachment protein) receptor) protein in a target cell. Examples of variant neurotoxin components may include a variant light chain of a botulinum toxin having one or more amino acid substitutions, modifications, deletions and / or additions. The variant light chain may have the same or better ability to prevent exocytosis, such as release of neurotransmitter vesicles. In addition, the biological effect of the variant may be reduced compared to the parent chemical entity. For example, a variant light chain of a botulinum toxin type A with an amino acid sequence removed may have a shorter biological persistence than the parent (or native) botulinum toxin type A light chain.

[0072] As used herein, the terms "improving epilepsy" or "improving epileptic seizures" refer to reducing the frequency of epileptic seizures and / or related symptoms, and / or reducing the severity of epileptic seizures (stabilization of the seizure state so that it does not progress from, for example, focal to generalized) and / or related symptoms, and / or maintaining a substantially seizure-free state in a subject, and / or ameliorating unwanted seizures associated with a disease or disorder, or any combination of the foregoing.

[0073] "Epileptic seizure" refers to any non-chronic or chronic seizure, including seizures caused by epilepsy, epilepsy, audiogenic epilepsy, epileptic syndromes, and related conditions. Seizures caused by fever, abnormal sodium or glucose blood levels, drug abuse, certain medications, hypertension, phenylketonuria, and uremia, and acute epilepsy such as those caused by brain tumors and / or brain damage, may turn into chronic epilepsy. The term "epilepsy" or "epilepsy" refers to a chronic neurological syndrome characterized by recurrent epileptic seizures.

[0074] The term "epilepsy" refers to those conditions involving chronic recurrent epileptic seizures, which can be considered as epilepsy. The term "epileptic seizure" should be understood to include any epileptic seizure state, whether it is part of epilepsy or as a result of another syndrome or related condition. Non-limiting examples of epileptic seizure types are focal epileptic seizures (also referred to as simple partial epileptic seizures, complex partial epileptic seizures, partial epileptic seizures with secondary generalized epileptic seizures, partial basic epileptic seizures, complex partial epileptic seizures or psychomotor epileptic seizures), generalized epileptic seizures (convulsive and non-convulsive, such as absence seizures, atypical absence seizures, myoclonic seizures, clonic seizures, tonic seizures, tonic-clonic seizures, atonic seizures (astatic), secondary generalized seizures and other chronic unclassified epileptic seizures.

[0075] As used herein, the term "partial seizure" refers to an epileptic seizure that affects a limited area of ​​the brain. In contrast, a "generalized seizure" is an epileptic seizure that affects the entire brain. Symptoms of partial seizures include, but are not limited to, abnormal muscle contractions, confusion, compulsive eye movements, abnormal sensations (e.g., numbness and tingling), hallucinations, abdominal pain, nausea, abnormal sweating, rapid heart rate, vision changes, mood changes, and loss of consciousness. Partial seizures are typically classified as "simple" or "complex" based on observed symptoms, electroencephalogram (EEG) data analysis, or other factors.

[0076] As used herein, the term "complex partial seizure" refers to a partial seizure that causes partial or total loss of consciousness. Complex partial seizures can occur in any area of ​​the brain. Complex partial seizures often occur in one of the two temporal lobes of the brain. A typical complex partial seizure lasts one to two minutes and is preceded by an "aura" (i.e., a warning feeling, such as nausea) that indicates an impending seizure.

[0077] As used herein, the terms "aura", "pre-seizure symptom" or "pre-epileptic aura symptom" refer to any warning sensation perceived by a patient that indicates an impending seizure or epilepsy, which may include, for example, nausea, visual changes, auditory changes, changes in smell, perception of strange lights, watery eyes, unpleasant smells (hallucinations) or tastes (hallucinations), synesthesia, head auras, confused thoughts or experiences, sudden feelings of anxiety, fear or dizziness, numbness or tingling, and hallucinations, etc.

[0078] As used herein, the term "simple partial seizure" refers to a partial seizure that does not cause loss of consciousness. A typical simple partial seizure lasts 60 seconds or less.

[0079] As used herein, the term "limbic seizure" refers to a seizure that originates in one of the limbic structures of the brain. The limbic system helps regulate involuntary movements of the body and hormonal activity. Limbic structures include, but are not limited to, the thalamus, hypothalamus, cingulate gyrus, amygdala, hippocampus, and basal ganglia.

[0080] As used herein, the term "chemically induced seizure" refers to a seizure in a subject caused by exposure of the subject to a chemical substance.

[0081] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to achieve the desired therapeutic effect.

[0082] As used herein, the term "in the vicinity of" means at or within about 1.5 cm, more preferably at or within about 1.0 cm and most preferably at or within about 0.5 cm of a specified reference location, for example, when botulinum toxin is administered according to the present disclosure, it is administered in the vicinity of at least one suture of a patient, i.e., at or within about 1.5 cm, more preferably at or within about 1.0 cm, or most preferably at or within 0.5 cm from a reference suture.

[0083] Less than the full measure of the disclosure may be claimed for any reason by reserving the right to proviso out or exclude any individual portion of any such group (including any subrange or combination of subranges within a group) as claimed, or in any similar manner. In addition, less than the full measure of the disclosure may be claimed for any reason by reserving the right to proviso out or exclude any individual substituent, analog thereof, compound thereof, ligand thereof, structure thereof, or group thereof, or any portion of a claimed group.

[0084] Throughout this disclosure, various patents, patent applications, and publications are cited. The entire disclosures of these patents, patent applications, and publications are incorporated herein by reference in their entirety to more fully describe the prior art known to those skilled in the art as of the date of this disclosure. If there are any inconsistencies between the cited patents, patent applications, and publications and this disclosure, the disclosure shall prevail.

[0085] For convenience, certain terms used in the specification, examples and claims are collected here. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0086] How to use

[0087] In one aspect, methods for treating epileptic seizures associated with epilepsy and for inhibiting the development of focal epileptic seizures into generalized epileptic seizures, methods for treating symptoms of premonitory epileptic seizures (or premonitory epileptic seizures), or methods for treating paroxysmal headaches are provided. The method comprises administering a therapeutically effective amount of a Clostridial derivative or a Clostridial toxin to a subject extracranially. A study was conducted that demonstrated that epileptic seizures treated with extracranially administered Clostridial derivatives in animals inhibited the development of focal epileptic seizures into generalized epileptic seizures and is described in Example 1. In the study, a first group of animals was treated with a Clostridial derivative prior to chemical induction of epileptic seizures. Prior to inducing chemical epileptic seizures, a second group of animals was treated with saline and a third group of animals did not receive prior treatment.

[0088] Regarding animals treated with Clostridium derivatives prior to chemically induced seizures, Figure 1A is a schematic diagram of a top view of an animal skull with a cut away to show the location of certain cranial sutures. The sagittal suture 10 is a connective tissue suture between the two parietal bones 12, 14. The lambdoid suture 16 is a connective tissue suture at the back of the skull that connects the parietal bones to the occipital bones 18. The coronal suture 20 is a connective tissue suture that separates the frontal bone 22 and the parietal bones 12 and 14. In the study described in Example 1, the Clostridium derivative was administered on or along the lambdoid sutures. The Clostridium derivative dose was divided into equal portions, five portions in this study, and one portion was administered by injection to five administration sites along the lambdoid sutures. The five administration sites were at Figure 1A The cells are marked with dots, such as represented by dot 24. Animals in the test group treated with the Clostridial derivative prior to seizures received doses of the Clostridial derivative 7-12 days prior to induction of chemical seizures with picrotoxin. Animals in the saline test group were treated similarly, except that each animal received saline instead of the Clostridial derivative at the administration site.

[0089] To induce epileptic seizures, Figure 1B Picrotoxin was applied topically at the location identified by the numerical identifier 30 in the . Electrodes 32, 34 for recording local field potentials and electrocorticograms were placed in the parietal and occipital cortices of all test animals. The electrocorticogram tracings are shown in Figures 2-3. Figure 2A In FIG, , tracings of the occipital cortex of the saline-treated group of animals as a function of time (in minutes) are shown. Tracings of the parietal cortex from saline-treated animals are shown in FIG. Figure 2B Immediately after time zero, picrotoxin was administered at a time determined by Figure 2A A few minutes after picrotoxin administration into the occipital cortex, focal epileptic seizures were induced in the occipital cortex ( Figure 2A ), which then develops to the parietal cortex ( Figure 2B ). Figure 2A-2B The traces in show that in untreated animals, induction of seizures with picrotoxin triggered focal seizures that progressed to generalized seizures.

[0090] Figure 3A Shown are tracings of the occipital cortex of animals in the group treated with Clostridial toxin prior to induction of epileptic seizures with picrotoxin. Picrotoxin was administered immediately after time zero, with the administration time being determined by Figure 3A A few minutes after picrotoxin administration into the occipital cortex, focal epileptic seizures were induced in the occipital cortex ( Figure 3A ). However, if Figure 3B As shown, focal epileptic seizures did not develop into generalized epileptic seizures. These observations indicate that administration of the Clostridium derivative inhibits or substantially prevents the development of focal epileptic seizures into generalized epileptic seizures.

[0091] In the above studies and Example 1, the Clostridium derivative was administered extracranially along the lambdoid suture. Figures 4A-4B It is a human skull ( Figure 4A ) and rat skull ( Figure 4B ) shows that the trigeminal nerve is not present in the cervical region where the clostridial derivative is administered. Thus, in one embodiment, the clostridial derivative is administered and / or each administration site does not include the trigeminal nerve. Extracranial administration is performed by injection into the scalp, which does not contain any muscle. The scalp is a layered structure, with the outermost layer being the skin (with sebaceous glands and hair follicles), followed by connective tissue, the parietal aponeurosis, loose slit connective tissue, and the pericranium. In one embodiment, the clostridial derivative is administered by injection into all layers of the scalp, and in other embodiments, it is administered into one, two, three, four, and / or five layers of the scalp.

[0092] In one embodiment, the clostridial derivative is applied to multiple administration sites including the scalp and / or cervical region. For example, the clostridial derivative can be applied to multiple administration sites in the occipital muscle and multiple administration sites in the paraspinal muscles of the neck and / or the scalp. In a more specific example, the clostridial derivative is applied to about 1-10, 1-8, 1-6, 2-6, 2-5 or 2-4 sites in the occipital muscle, and / or 2-6, 2-4 sites in the paraspinal muscles of the neck, and / or 1-8, 1-6, 2-6, 3-5 sites in the scalp, which may be included in the seam or seam joint or along the seam or seam joint. Administration can be by injection, for example, intramuscular injection, intradermal injection and / or subcutaneous injection. In some embodiments, ideally, administration delivers the clostridial derivative to the cervical nerve. In one embodiment, at least one administration site is to the posterior head domain of the skull innervated by the cervical nerve.

[0093] In one embodiment, at least one administration site is near and / or generally along any suture of the skull, such as Figure 5A , 5B and 5C for example. Figure 5A is a front perspective view of a human skull 11, Figure 5B is a rear view of a human skull 11 . Figure 5C Also shown are some less common sutures that have been identified in the human skull. The bones that make up the skull include the frontal bone 13, parietal bone 15, temporal bone 17, sphenoid bone 19, and occipital bone 21. The edges of these bones are held together by sutures. For example, the coronal suture 23 (at Figure 5C The coronalis is located at the junction of the frontal bone 13 and the parietal bone 15. Figure 5C The lambdoid suture 26 (also called the squamosa) is located at the junction between the parietal bone 15 and the temporal bone 17. Figure 5C The sagittal suture 28 (also called the lambdoidea) is located at the junction between the parietal bone 15 and the occipital bone 21. Figure 5C ) at the connection between the two parietal bones, and at the connection between the frontal bone 13 and the nasal bone 33 of the frontonasal suture 31.

[0094] Thus, a method for treating epileptic seizures and / or for inhibiting or preventing the development of epileptic seizures and / or inhibiting or preventing secondary spread from an epileptic seizure focus is contemplated. The method comprises administering a Clostridium derivative to a subject in need thereof. In one embodiment, the Clostridium derivative is administered extracranially. In some embodiments, it is administered extracranially to the scalp. In some embodiments, it is administered to at least one suture in the skull. In other embodiments, it is administered to the trigeminal nerve in the skull. In other embodiments, it is administered to a muscle in the skull.

[0095] In some embodiments, it is applied to at least one suture in the cervical region. In other embodiments, it is applied to the trigeminal nerve in the cervical region. In other embodiments, it is applied to the muscles in the cervical region.

[0096] In other embodiments, the clostridial derivative is applied by extramuscular injection, subcutaneous injection, subdermal injection or intradermal injection. The clostridial derivative can be applied to a site or more than one site or multiple sites. In one embodiment, the clostridial derivative is applied to about 2-15 sites, about 2-12 sites, about 2-10 sites, about 2-8 sites, about 2-6 sites, about 2-5 sites, about 3-6 sites, about 3-5 sites, or is applied to 3,4,5,6,7,8,9 or 10 sites. In one embodiment, the site is an injection site.

[0097] As described above, the Clostridium derivative is administered extracranial. It is contemplated that the drug is administered to one or more sites on or adjacent to the cranial sutures, such as the lambdoid suture, sagittal suture, coronal suture, squamous suture. The Clostridium derivative can be administered to sutures and / or suture junctions, such as the junction of the lambdoid suture and the sagittal suture, or the junction of the coronal suture and the sagittal suture. In one embodiment, the Clostridium derivative is administered to the occipital region of the skull. In the occipital region, administration can be to the area innervated by the cervical nerves.

[0098] In some embodiments, the Clostridium derivative is administered to the primary region of the epileptic seizure. In another embodiment, the Clostridium derivative is administered contralateral to the primary region of the epileptic seizure.

[0099] In some embodiments, the Clostridium derivative is botulinum toxin type A. In some embodiments, the Clostridium derivative is selected from the group consisting of onabotulinumtoxinA, abobotulinumtoxinA, incobotulinumtoxinA, daxibotulinumtoxinA, prabotulinumtoxinA, and combinations thereof. In one embodiment, botulinum toxin type A can be applied to the sagittal suture at about 5-50 units divided into 2-6 sites; to the coronal suture at about 10-60 units divided into 2-8 sites; to the temporalis and / or squamous suture at about 10-100 units divided into 2-20 sites; to the occipital and / or lambdoid suture at about 10-60 units divided into 2-10 sites; and to at least one of the cervical paravertebral sites at about 1-20 units divided into 1 site or 2-4 sites, to the corrugator supercilii at about 1-30 units divided into 2-6 sites, and to the procerus at about 1-10 units divided into 1-4 sites. In one embodiment, the treatment paradigm further comprises applying botulinum toxin type A at a dose of about 5 units at 1 site at the lambdoid-sagittal junction.

[0100] Other examples include administering botulinum toxin type A at about 15 units to 3 sites at the sagittal suture; at about 50 units to 5 sites at the coronal suture; at about 30 units to 6 sites at the squamous suture; at about 30 units to 6 sites at the lambdoid suture; and / or at about 1 unit to 1 site at the lambdoid-sagittal junction.

[0101] In one embodiment, the epileptic seizures experienced by the subject to be treated are associated with a disorder or condition. Non-limiting examples include epilepsy, juvenile myoclonic epilepsy, idiopathic generalized epilepsy, and Lennox-Gastaut syndrome. In another embodiment, the epileptic seizures experienced by the subject to be treated are associated with an epileptic syndrome, including epilepsy or other disorders characterized by epileptic seizures. Non-limiting examples of epilepsy syndromes and other conditions associated with epileptic seizures include benign familial neonatal seizures, early myoclonic encephalopathy, Ohtahara syndrome, migrating partial seizures of infancy, West syndrome, benign myoclonic epilepsy in infancy, benign familial infantile seizures, benign infantile seizures (non-familial), Dravet syndrome, hemiconvulsive-hemiplegic syndrome, benign childhood epilepsy with centrotemporal spikes, early-onset benign childhood occipital epilepsy (Panayiotopoulos type), late-onset benign childhood occipital epilepsy (Gastaut type), myoclonic absence epilepsy, epilepsy with myoclonic-astatic seizures, myoclonic-astatic seizures, Lennox-Gastaut syndrome, Landau-Kleffner syndrome (LKS), sustained spike-wave epilepsy during slow-wave sleep (except LKS), childhood absence epilepsy, progressive myoclonus epilepsies, idiopathic generalized epilepsy with variable phenotype, juvenile absence epilepsy, juvenile myoclonic epilepsy, epilepsy with generalized tonic-clonic seizures only, reflex epilepsy, idiopathic photosensitive occipital lobe epilepsy, startle epilepsyepilepsy, autosomal dominant nocturnal frontal lobe epilepsy, familial temporal lobe epilepsy, generalized epilepsy with febrile seizures plus, familial focal epilepsy with variable foci, symptomatic focal epilepsies, limbic epilepsies, mesial temporal lobe epilepsy with hippocampal sclerosis, mesial temporal lobe epilepsy defined by specificetiologies.

[0102] The purposes of the method herein are also applicable to treating epileptic seizures and / or inhibiting the development of epileptic seizures in patients experiencing epileptic seizures unrelated to epilepsy. The condition of epileptic seizures that do not require epilepsy diagnosis includes, for example, benign neonatal seizures, febrile seizures, reflex seizures, alcohol withdrawal seizures, seizures induced by drugs or other chemicals, immediate or early post-traumatic seizures, single seizures or isolated clusters of seizures, rarely repeated seizures (oligoepilepsy). In one embodiment, the epileptic seizure to be treated is a partial onset seizure, a myoclonic seizure, or a primary generalized tonic-clonic seizure. In another embodiment, the seizure is a generalized tonic-clonic seizure associated with Lennox-Gastaut syndrome.

[0103] The methods described herein may be adjunct to oral, subcutaneous, rectal or intravenous administration of medications known in the art for stopping or preventing epileptic seizures.

[0104] Clostridium derivatives

[0105] In one embodiment, the Clostridium derivative is a botulinum neurotoxin (BoNT), e.g., BoNT / A, BoNT / B, etc. These toxins act on the nervous system by blocking the release of neurosecretory substances such as neurotransmitters. The action of BoNT is initiated by its binding to receptor molecules on the cell surface, and the toxin-receptor complex then undergoes endocytosis. Once inside the cell, the BoNT cleaves the exocytosis-specific proteins responsible for neurotransmitter docking and releases them from the cell, called SNARE proteins (soluble N-ethylmaleimide-sensitive factor attachment protein receptors). The resulting transient chemodenervation has been used medically to block motor neurotransmission at the neuromuscular junction, leading to a variety of therapeutic applications.

[0106] In some embodiments, the Clostridium derivative comprises a natural recombinant Clostridium toxin, a recombinant modified toxin, a fragment thereof, a TEM, or a combination thereof. In some embodiments, the Clostridium derivative is a botulinum toxin. In some embodiments, the botulinum toxin can be a botulinum toxin of type A, type B, type C1, type D, type E, type F, or type G, type H, type X, a chimeric botulinum toxin, or any combination thereof. The botulinum neurotoxin can be a recombinantly prepared botulinum neurotoxin, such as a botulinum toxin produced by Escherichia coli. In alternative embodiments, the Clostridium derivative is a TEM.

[0107] In some embodiments, the botulinum neurotoxin may be a modified neurotoxin, i.e., a botulinum neurotoxin in which at least one amino acid is deleted, modified or substituted compared to the natural toxin, or the modified botulinum neurotoxin may be a recombinantly produced botulinum neurotoxin or a derivative or fragment thereof. In certain embodiments, the modified toxin has an altered cell targeting ability for target neurons or non-neuronal cells. This altered ability is achieved by replacing the naturally occurring targeting domain of botulinum toxin with a targeting domain that exhibits selective binding activity to a non-botulinum toxin receptor present in a non-botulinum toxin target cell. This modification of the targeting domain results in the ability of the modified toxin to selectively bind to a non-botulinum toxin receptor (target receptor) present in a non-botulinum toxin target cell (retargeted). The modified botulinum toxin having targeting activity for non-botulinum toxin target cells can bind to receptors present on non-botulinum toxin target cells, be transferred to the cytoplasm, and exert its proteolytic effect on the SNARE complex of the target cell. In essence, by selecting an appropriate targeting domain, the botulinum toxin light chain comprising an enzyme domain is delivered intracellularly to any desired cell.

[0108] The clostridial derivatives such as botulinum toxin used in accordance with the methods of the present invention can be stored in a freeze-dried, vacuum-dried form in a container under vacuum pressure or as a stable liquid. Prior to freeze-drying, the botulinum toxin can be combined with a pharmaceutically acceptable excipient, stabilizer and / or carrier such as albumin, etc. In embodiments containing albumin, the albumin can be, for example, human serum albumin, etc. The freeze-dried material can be reconstituted with a suitable liquid such as saline, water, etc. to form a solution or composition containing the botulinum toxin to be administered to a patient.

[0109] In some embodiments, the clostridial derivative is provided in a controlled release system comprising a polymer matrix encapsulating the clostridial derivative, wherein a portion of the clostridial derivative is released from the polymer matrix in a controlled manner over an extended period of time. Controlled release neurotoxin systems have been disclosed, for example, in U.S. Pat. Nos. 6,585,993; 6,585,993; 6,306,423 and 6,312,708, each of which is incorporated herein by reference in its entirety.

[0110] In alternative embodiments, the Clostridium derivative is provided as an ointment, gel, cream or emulsion suitable for topical administration.

[0111] The therapeutically effective amount of the clostridial derivative (e.g., botulinum toxin) administered according to the method of the present invention can vary according to the efficacy of the toxin and the specific characteristics of the pain being treated, including its severity and other various patient variables, including stature, weight, age, and responsiveness to treatment. The efficacy of the toxin is expressed as a multiple of the mouse LD50 value, and one unit (U) of the toxin is defined as the corresponding amount of the toxin that kills 50% of a group of 18 to 20 female Swiss-Webster mice (Swiss-Webstermice) (each weighing about 20 grams). The efficacy of the clostridial derivative can also be expressed in mass units. The effective amount expressed in mass units can be determined based on the expected effect. For example, the effective amount can be determined based on the amount of the clostridial derivative required to relieve epileptic seizures or inhibit epileptic seizures from developing from focal epileptic seizures to generalized epileptic seizures, and not producing significant toxicity.

[0112] The therapeutically effective amount of botulinum toxin can vary depending on the potency of the particular botulinum toxin, as commercially available botulinum toxin preparations do not have equivalent units of potency. A unit of onabotulinumtoxinA (a botulinum toxin type A available from Allergan) is approximately equal to 3 to 5 units of (abobotulinumtoxinA) (also botulinum toxin type A available from Ipsen Pharmaceuticals). (known outside the United States as ), a botulinum toxin type B available from Elan (currently USWorldmeds), reportedly has a potency unit relative to In some embodiments, the botulinum neurotoxin can be a pure toxin without complexing proteins, such as (incobulinumtoxinA). Therefore, the amount of toxin administered and the frequency of its administration will be determined by the treating physician and will be commensurate with the safety issues and effects of the particular toxin formulation. In an embodiment, the Clostridium derivative is selected from onabotulinumtoxinA, incobulinumtoxinA, abotulinumtoxinA, daxibotulinumtoxinA, prabotulinumtoxinA and rimabotulinumtoxinB.

[0113] Where more than one administration site is desired, the dose of the Clostridium derivative to be administered may be divided evenly for administration to each administration site. It is also contemplated that the dose to be administered may be divided into unequal portions for administration to each administration site.

[0114] In one embodiment, the therapeutic effect of the Clostridium derivative lasts for about 1 month to 5 years, or about 1 month to 1 year, or about 1 month to 6 months, or about 1 month to 5 months, about 1 month to 4 months, about 1 month to 3 months, about 1 month to 2 months. In some embodiments, treatment can be provided continuously or intermittently as needed.

[0115] In one embodiment, the clostridial derivative is present in a composition, which is a stable liquid or solid pharmaceutical composition. An exemplary composition comprises a clostridial derivative, a disaccharide, a surfactant and an antioxidant. The composition may include a pharmacologically acceptable excipient. As used herein, "pharmacologically acceptable excipient" is synonymous with "pharmacological excipient" or "excipient", and refers to any excipient that has substantially no long-term or permanent harmful effects when applied to a mammal, and includes compounds such as stabilizers, fillers, cryoprotectants, lyoprotectants, additives, excipients, carriers, diluents or adjuvants. Excipients are usually mixed with active ingredients or allow dilution or encapsulation of active ingredients, and may be solid, semisolid or liquid agents. It is also envisioned that a pharmaceutical composition comprising a clostridial derivative may include one or more pharmaceutically acceptable excipients, which help to process the active ingredient into a pharmaceutically acceptable composition. To the extent that any pharmacologically acceptable excipient is not incompatible with the clostridial derivative, it is considered to be used in a pharmaceutically acceptable composition. Non-limiting examples of pharmacologically acceptable excipients can be found, for example, in Pharmaceutical Dosage Forms and Drug Delivery Systems (Howard C. Ansel et al., eds., Lippincott Williams & Wilkins, 7th edition, 1999); Remington: The Science and Practice of Pharmacy (Alfonso R. Gennaro, ed., Lippincott, Williams & Wilkins, 20th edition, 2000); Goodman & Gilman's The Pharmacological Basis of Therapeutics (Joel G. Hardman et al., eds., McGraw-Hill Professional, 10th edition, 2001); and Handbook of Pharmaceutical Excipients (Raymond C. Rowe et al., APhA, 4th edition, 2003), the entire contents of each of which are incorporated herein by reference.

[0116] The components of the pharmaceutical composition may be contained in a single composition (i.e., all components are present at the time of initial mixing of the pharmaceutical composition, except for any required reconstitution fluid) or as a two-component system, such as a vacuum-dried composition reconstituted with a reconstitution vehicle, which may, for example, contain components that were not present in the initial mixing of the pharmaceutical composition. Two-component systems can provide several benefits, including allowing the incorporation of components that are not fully compatible with the first composition of the two-component system for long-term shelf storage. For example, the reconstitution vehicle may include a preservative that provides adequate protection against microbial growth during use (e.g., one week of refrigerated storage), but is absent during two years of frozen storage, during which it may decompose the toxin. Other ingredients that may not be compatible with botulinum toxin or other ingredients for a long time may be incorporated in this manner; i.e., incorporated into the second vehicle (e.g., the reconstitution vehicle) at the approximate time of use. The pharmaceutical composition may also include preservatives such as benzyl alcohol, benzoic acid, phenol, parabens, and sorbic acid. The pharmaceutical composition may contain, for example, excipients such as surfactants; dispersants; inert diluents; granulating and disintegrating agents; binders; lubricants; preservatives; physiologically degradable compositions such as gelatin; aqueous excipients and solvents; oily excipients and solvents; suspending agents; dispersants or wetting agents; emulsifiers; demulcents; buffers; salts; thickeners; fillers; antioxidants; stabilizers; and pharmaceutically acceptable polymeric or hydrophobic materials and other active ingredients known in the art and described, for example, in Genaro, ed., 1985, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., which are incorporated herein by reference.

[0117] In embodiments, the botulinum toxin may be selected from botulinum toxin types A, B, C, D, E, F, G, H, X, and chimeric toxins. In some embodiments, the botulinum toxin is botulinum toxin type A. The botulinum toxin may be administered according to a treatment paradigm to deliver to the patient a total amount of about 1 unit to about 3000 units, or about 2 units to about 2000 units, or about 5 units to about 1000 units, or about 10 units to about 500 units, or about 15 units to about 250 units, or about 20 units to about 150 units, or 25 units to about 100 units, or about 30 units to about 75 units, or about 35 units to about 50 units. In some embodiments, the Clostridium derivative can be administered at a dose of about 0.001 ng to about 1000 ng, preferably about 0.001 ng to about 500 ng, preferably about 0.01 ng to about 250 ng, preferably about 0.1 ng to about 150 ng, preferably about 1 ng to about 100 ng, preferably about 1 ng to about 10 ng. For example, onabotulinumtoxin A can be administered at a dose of about 1 ng, 2 ng, 3 ng, 4 ng, 5 ng, 6 ng, 7 ng, 8 ng, 9 ng or 10 ng. In some embodiments, the relief of symptoms can last from about 1 month to about 5 years.

[0118] In one embodiment, the clostridial derivative is administered in a sterile, vacuum-dried state for reconstitution in the form of a composition comprising about 100 units of botulinum neurotoxin type A complex, 0.5 mg human serum albumin, and 0.9 mg sodium chloride. In other embodiments, the clostridial derivative is administered in a composition that does not contain animal protein. In some embodiments, the clostridial derivative is administered in a composition that does not contain animal protein, comprising a surfactant and a sugar. In some embodiments, the clostridial derivative is administered in a composition that does not contain animal protein, comprising botulinum toxin type A, poloxamer, and trehalose or sucrose. In one embodiment, the clostridial derivative is administered in a sterile, vacuum-dried state for reconstitution in the form of a composition that comprises botulinum neurotoxin type A complex, 8% trehalose, 4% poloxamer 188, and 0.2% methionine.

[0119] The step of administering a clostridial derivative, such as type A botulinum toxin, can include using a needle to penetrate one or more layers of the scalp and insert near and along the patient's cranial sutures, then gradually withdrawing the needle along the sutures while delivering the toxin along the sutures. In some embodiments, a botulinum toxin, such as type A or type B botulinum toxin, is administered along the sutures using more than one injection site. Administration can be performed substantially along the length of the suture, such as at or up to about 1.5 cm from the reference suture, more preferably at or up to about 1.0 cm from the reference suture, and most preferably at or up to about 0.5 cm from the reference suture, and used as a guide for administration of botulinum toxin according to the present disclosure. In one embodiment, a syringe equipped with a needle and filled with a composition comprising a clostridial toxin is provided, and the needle is inserted along the suture below the surface of the skin to administer the toxin. Example

[0120] The following non-limiting examples provide those of ordinary skill in the art with specific preferred methods within the scope of the embodiments of the present method and are not intended to limit the scope thereof.

[0121] Example 1

[0122] Treatment with Clostridial toxins inhibits the progression of epileptic seizures

[0123] The experimental animals (rats) were randomly divided into three groups for treatment. The first group of animals was not treated. The second group was treated with clostridial toxin onabotulinumtoxinA, which entered the scalp area innervated by the greater and lesser occipital nerves. 7-12 days before inducing epilepsy, 5 units of onabotulinumtoxinA were applied to 5 administration sites, and 1 unit was given at each site. The third group of animals received saline injections in the same area injected with onabotulinumtoxinA, also 7-12 days before inducing epilepsy.

[0124] Glass pipette electrodes capable of recording local field potentials and electrocortical maps from 0 to 3000 Hz were used to identify cortical activity before and during epileptic seizures induced by topical application of picrotoxin, a selective GABA blocker. Figure 1B shown.

[0125] Figure 2A-2BElectrocortical electrograms of the occipital and parietal cortices of the second and third groups of animals (clostridial toxin-treated and saline-treated, respectively) are shown in 3A-3B before and after picrotoxin A was applied topically to the occipital cortex to induce focal epileptic seizures. This study provides evidence that the development and progression of epileptic seizures from focal to generalized can be prevented by extracranial injection of onabotulinumtoxinA into the scalp areas innervated by the greater and lesser occipital nerves (both originating from the C2 and C3 dorsal root ganglia, respectively).

[0126] Example 2

[0127] Treatment of post-traumatic epilepsy with clostridial toxins

[0128] A 42-year-old woman was an unrestrained passenger in a head-on motor vehicle collision. She sustained head injuries resulting in skull fractures, bifrontal subdural hematomas, left frontal cortical contusions, and left frontal intraprenchymal hemorrhage. After three weeks of intensive care, she was transferred to the medical ward and from there to a rehabilitation facility. Although she did not have epilepsy, she was at risk for a post-traumatic seizure in the lower ear (~20% probability).

[0129] The patient did not want to take any oral medications. She did not have any seizures after the accident. However, 9 months after the accident, she developed an episode of expressive aphasia that progressed to tonic / clonic (TC) seizures in her right arm and leg within 60 seconds and then generalized to bilateral TC seizures with loss of consciousness 30 seconds later. After one such seizure that did not resolve spontaneously, she was admitted to the emergency department for intravenous (IV) anti-epileptic medication administration. After recovery, she had difficulty with expressive language. On continuous monitoring, the EEG showed paroxysmal focal spike slowing over the left frontal lobe associated with expressive aphasia. She was started on oral medications, which had to be changed regularly because they were either not tolerated or did not provide adequate seizure relief. Her best seizure control was on lamotrigine, and although the dose was optimized, she still had episodes of expressive aphasia that progressed to right TC spasms and generalized, but were short-lived. These occurred approximately monthly.

[0130] She was treated with onabotulinumtoxinA as follows. Five units were injected into each of four sites in the frontalis muscle, one injection of 10 units near the frontonasal suture, seven scattered injections of 5 units near the coronal suture (subfascial) (distributed over the entire length of the suture), and six injections of 5 units into the left temporalis muscle, including two injections near the squamous suture above and in front of the ear and one injection near the squamous suture posteriorly. There were three subfascial injections of 5 units distributed along the sagittal suture. She had episodes of expressive aphasia that lasted approximately 10 seconds, but did not progress to the right and were not generalized. She discontinued lamotrigine. She continued treatment every 3-5 months with brief episodes of expressive aphasia occurring monthly.

[0131] Example 3

[0132] Treatment with Clostridial toxins reduces the occurrence of epileptic seizures

[0133] An adult female has several epileptic seizures per week, characterized by the sudden onset of confusion with difficulty understanding language for about 20 seconds, followed by loss of consciousness and convulsions lasting 1 minute. The diagnostic results (EEG and MRI) showed that the onset of the epileptic seizure was located in the left temporoparietal region, overlapping with the cortical receptive language area. Therefore, the patient's epileptic seizure focus is located in an area that is inaccessible to epilepsy surgery. Despite the trial of multiple anti-epileptic drugs, including levetiracetam, oxcarbazepine, and lamotrigine, the epileptic seizures continued. Therefore, the patient suffers from epilepsy that is drug-resistant and inoperable.

[0134] The patient's left temporoparietal region was treated unilaterally with a total dose of 40 units Fifteen units were administered at three sites near the lambdoid suture. Ten units were administered at two sites near the parieto-mammary suture, and 15 units were administered at three sites near the occipital suture. Within one week of treatment, the patient did not have any seizures. Over the next three months, the patient had only three seizures. During the fourth month, she experienced three seizures and returned for further treatment. She was retreated every three months with the same treatment pattern and remained seizure-free during the treatment period.

[0135] Example 4

[0136] Treatment with Clostridial toxins to relieve generalized epileptic seizures

[0137] A 4-year-old boy with autism and developmental delay presented with frequent brief tonic seizures. His MRI was normal, and his EEG showed frequent slow spikes and waves consistent with Lennox-Gastaut syndrome. Despite a trial of multiple anti-epileptic drugs and a ketogenic diet, the seizures persisted.

[0138] The patient was treated bilaterally with 5 units of onabotulinumtoxinA per administration site as follows: 3 injections into each temporalis muscle, 2 injections into each occipital muscle, 2 injections along the parieto-mammary suture on each side, and 2 injections into the paraspinal muscles of the neck innervated by the cervical nerve on each side, for a total of 18 injections for a total of 90 units. Over the next 3 months, the patient experienced a 50% decrease in the frequency of seizures. At the end of 3 months, the patient was retreated using the same regimen with the following additional sites: 4 injections along the coronal suture, 3 injections along the sagittal suture, 1 injection along the posterior squamous suture on each side, and 2 injections along the lambdoid suture on each side. The additional treatment provided an additional 13 injections for an additional 65 units. The total treatment was 155 units. The patient was treated with this paradigm every 3 months and remained seizure-free.

[0139] Example 5

[0140] Treatment with Clostridial toxin reduces focal epileptic seizures associated with brain tumors

[0141] A 35-year-old man with a glioblastoma multiforme that originated in the right hemisphere but spread to the left through the corpus callosum presented with focal seizures that occurred independently in the left and right hemispheres. Despite a trial of multiple anti-epileptic drugs, the seizures persisted. However, based on the EEG, the findings were more prominent on the right side.

[0142] The patient was treated with onabotulinumtoxinA, 10 U per injection, as follows: 4 injections along the coronal suture (2 per side), 3 injections along the sagittal suture, and 3 injections along the right lambdoid suture. The total amount administered was 100 U. Interestingly, the EEG spike activity was significantly suppressed bilaterally, and although his EEG continued to be abnormal, he had no clinical seizure activity.

[0143] Without departing from the spirit and scope of the present disclosure, a person of ordinary skill in the art may make many changes and modifications. Therefore, it must be understood that the described embodiments are set forth for the purpose of example only, and these embodiments should not be considered to limit the scope of the appended claims. Therefore, the appended claims should be understood to include not only the combination of elements literally set forth, but also all equivalent elements that perform substantially the same function in substantially the same manner to obtain substantially the same results. Therefore, the claims should be understood to include those already described above, those that are conceptually equivalent, and those that are incorporated into the present disclosure.

Claims

1. A method for treating epileptic seizures associated with epilepsy or for treating symptoms of premonitory epileptic seizures, comprising: A Clostridium derivative is administered to a subject having epilepsy or having symptoms of a premonitory seizure in an amount effective to treat the seizure, wherein the Clostridium derivative is administered extracranially to at least one administration site.

2. The method of claim 1, wherein administering comprises administering by extramuscular injection, subcutaneous injection, subdermal injection, or intradermal injection.

3. The method of claim 1, wherein administering comprises administering to the scalp of the subject by injection.

4. The method of any one of claims 1-3, wherein the administering is to more than one administration site.

5. The method of claim 4, wherein the administering is to 2-20 administration sites.

6. The method of claim 5, wherein the administration site is approximately along any suture line on the subject's skull or approximately along the lambdoid suture.

7. The method of any of the preceding claims, wherein the at least one administration site is approximately at the junction of the lambdoid and sagittal sutures.

8. The method of any of the preceding claims, wherein the at least one administration site is located in the posterior head region of the skull innervated by the cervical nerves.

9. The method of any one of claims 4-7, wherein the amount of the Clostridium derivative administered is evenly divided into several portions for administration to each administration site.

10. The method of any one of claims 4-7, wherein a portion of the amount of the Clostridium derivative is administered to each administration site per injection site, wherein the portion administered at each administration site is unequal.

11. The method of any of the preceding claims, wherein the Clostridium derivative is a botulinum neurotoxin.

12. The method of claim 11, wherein the amount administered is about 1-3000 units.

13. The method of claim 11, wherein the botulinum neurotoxin is botulinum toxin type A, botulinum toxin type B, botulinum toxin type C, botulinum toxin type D, or botulinum toxin type E.

14. The method of claim 11 or claim 13, wherein the botulinum toxin is selected from onabotulinumtoxinA, abobotulinumtoxinA, incobotulinumtoxinA, daxibotulinumtoxinA, prabotulinumtoxinA and rimabotulinumtoxinB.

15. The method of claim 12 or claim 13, wherein the botulinum toxin comprises a botulinum neurotoxin complex or pure toxin.

16. The method of claim 12, wherein the administered botulinum toxin is evenly distributed among about 2-10 administration sites.

17. The method of any of the preceding claims, wherein the seizure is a focal seizure, a myoclonic seizure, or a primary generalized tonic-clonic seizure.

18. The method of claim 17, wherein the seizure is associated with an epilepsy disorder selected from the group consisting of epilepsy, juvenile myoclonic epilepsy, idiopathic generalized epilepsy, and Lennox-Gastaut syndrome.

19. The method of any one of claims 1-16, wherein the seizure is a generalized tonic-clonic seizure associated with Lennox-Gastaut syndrome.

20. The method of any of the preceding claims, wherein the administration is adjunctive therapy to oral or intravenous administration for the treatment or control of epileptic seizures.

21. A method for inhibiting the progression of a focal epileptic seizure to a generalized epileptic seizure, comprising: A therapeutically effective amount of a Clostridium derivative is administered extracranially to the scalp of a subject suffering from focal epileptic seizures to inhibit the progression of the focal epileptic seizures to generalized epileptic seizures.

22. A method for treating seizures associated with epilepsy, comprising: A therapeutically effective amount of a Clostridium derivative is administered extracranially to the scalp of a subject suffering from epileptic seizures to treat the epileptic seizures.

23. A method for treating an epileptic seizure in a subject suffering from epilepsy or for treating a premonitory epileptic seizure (or premonitory epileptic seizure) symptom in a subject suffering from premonitory epileptic seizure (or premonitory epileptic seizure) symptom, comprising: A therapeutically effective amount of a Clostridium derivative is administered extracranially to the scalp of a subject to treat epileptic seizures.

24. A method for controlling focal epileptic seizures, comprising: The Clostridium derivative is administered extracranially to the subject in an amount effective to control focal epileptic seizures to a therapeutically effective amount.

25. A method for preventing a focal asymptomatic epileptic seizure from developing into a focal symptomatic epileptic seizure, comprising: An effective amount to a therapeutically effective amount of the Clostridium derivative is administered extracranially to a subject to prevent the progression of a focal asymptomatic seizure to a focal symptomatic seizure.

26. The method of any one of claims 21-25, wherein administering comprises administering by extramuscular injection, subcutaneous injection, subdermal injection, or intradermal injection.

27. The method of any one of claims 21-25, wherein administering comprises administering to the scalp of the subject by injection.

28. The method of any one of claims 21-27, wherein administration is to more than one administration site.

29. The method of claim 28, wherein administering is to 2-20 administration sites.

30. The method of claim 29, wherein the administration site is generally along any suture line in the skull or generally along the lambdoid suture.

31. The method of any of the preceding claims, wherein the at least one administration site is approximately at the junction of the lambdoid and sagittal sutures.

32. The method of any of the preceding claims, wherein the at least one administration site is located in the posterior head region of the skull innervated by the cervical nerves.

33. The method of any one of claims 28-31, wherein the amount of the Clostridium derivative administered is evenly divided into portions for administration to each administration site.

34. The method of any one of claims 28-31, wherein a portion of the amount of the Clostridium derivative is administered to each administration site per injection site, wherein the portion administered at each administration site is unequal.

35. The method of any of the preceding claims, wherein the Clostridium derivative is a botulinum neurotoxin.

36. The method of claim 35, wherein the amount administered is about 1-3000 units.

37. The method of claim 35, wherein the botulinum neurotoxin is botulinum toxin type A, botulinum toxin type B, botulinum toxin type C, botulinum toxin type D, or botulinum toxin type E.

38. The method of claim 35 or claim 37, wherein the botulinum toxin is selected from onabotulinumtoxinA, abobotulinumtoxinA, incobotulinumtoxinA, daxibotulinumtoxinA, prabotulinumtoxinA and rimabotulinumtoxinB.

39. The method of claim 36 or claim 37, wherein the botulinum toxin comprises a botulinum neurotoxin complex or pure toxin.

40. The method of claim 36, wherein the administered botulinum toxin is evenly distributed among about 2-10 administration sites.

41. The method of any of the preceding claims, wherein the administration is adjunctive therapy to oral or intravenous administration for the treatment or control of epileptic seizures.

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