A method for the treatment of epilepsy

Abstract

The present disclosure provides a method for the treatment of epilepsy syndrome and / or seizure in a subject, by administering a therapeutically effective amount of the compound or salt thereof as disclosed herein. Embodiments herein further overcome the challenges that are generally observed with the use of existing anti- epileptic drugs, such as poor patient response, severe physical and / or psychological side effects and prevalence of drug resistance, etc. Accordingly, the present disclosure provides a compound for use in treating epilepsy syndrome and / or seizure. Also disclosed herein is a composition for the treatment of epilepsy syndrome and / or seizure.

Description

[0001] A METHOD FOR THE TREATMENT OF EPILEPSY

[0002] FIELD OF INVENTION

[0003]

[0001] The present disclosure relates to a method for the treatment of epilepsy syndrome and / or seizure in a subject. Particularly, the disclosure provides compositions and compounds for use in treating epilepsy syndrome and / or seizure in a subject.

[0004] BACKGROUND OF THE INVENTION

[0005]

[0002] Epilepsy is the fourth most common neurological disease, affecting 45 million people worldwide ]Beghi, E., The epidemiology of Epilepsy. Neuroepidemiology, 2020. 54(2): p. 185-191]. Epilepsy is characterized by unprovoked, synchronous, and repetitive seizures caused by imbalances in excitatory and / or inhibitory neurotransmission (E / I) [Falco-Walter, J. J., I.E. Scheffer, andR.S. Fisher, The new definition and classification of seizures and Epilepsy. Epilepsy research, 2018. 139: p. 73-79]. The disease comprises 3 phases: I- latency phase or epileptogenesis (modifications of brain network required for increased susceptibility to seizures); II- spontaneous seizures phase marked by the first appearance of first unprovoked seizures, and lastly, III- chronic seizure phase or maturation phase marked by increased frequency and intensity of the seizures [Reddy, D.S., Role of hormones and neurosteroids in Epileptogenesis. Frontiers in cellular neuroscience, 2013. 7: p. 115].

[0006]

[0003] Present treatment options include anti-epileptic drugs (AEDs) and surgical intervention. Generally used AEDs include carbamezapine, diazepam, ethosuximide, felbamate, ketogenic diet, lacosamide, lamotrigine, levetiracetam, lorazepam, midazolam, N-desmethylclobazam, nordiazepam, clobazam, clonazepam, clonidine, clorazepate, topiramate, trazodone, etc. Despite the availability of anti-epileptic drugs (AEDs) in the market, treatment challenges remain as: 1) More than 30% of the patients do not respond to the mono- or polytherapy of AEDs, and 2) Drug responders are presented with tremendous physical and / or psychological side effects [Simonato, M., et al., The challenge and promise of anti-epileptic therapy development in animal models. The Lancet Neurology, 2014. 13(9): p. 949-960]. Further, the currently available anti-epileptic drugs target the epileptic effectors-proteins that set the seizure threshold, but not the epileptic mediators which are downstream signaling cascades contributing to the pathogenesis and severity of seizures.

[0007]

[0004] Therefore, there is a need for more tolerable treatments to act as homeostatic boosters, which targets the mediators of epilepsy thereby, preventing or blocking the development of the chronic phase of epilepsy by modifying the neural network from pathological to a near-physiological state.

[0008] SUMMARY OF THE INVENTION

[0009]

[0005] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the disclosed subject matter, nor is it intended to be used to limit the scope of the disclosed subject matter.

[0006] In an aspect of the present disclosure, there is provided a method for the treatment of epilepsy syndrome and / or seizure in a subject, comprising administering a therapeutically effective amount of a compound, or salt thereof, of Formula I,

[0010] Formula I wherein A, B, C, X, Y, and Z are independently selected from H or OH groups.

[0011]

[0007] In another aspect of the present disclosure, there is provided a compound, or salts thereof, of Formula I, wherein A, B, C, X, Y, and Z are independently selected from H or OH groups, for use in the treatment of epilepsy syndrome and / or seizure in a subject.

[0012]

[0008] In yet another aspect of the present disclosure, there is provided a composition comprising a compound, or salts thereof, of Formula I, wherein A, B, C, X, Y, and Z are independently selected from H or OH groups; and at least one pharmaceutically acceptable carrier, for use in the treatment of epilepsy syndrome and / or seizure in a subject.

[0013] BRIEF DESCRIPTION OF DRAWINGS

[0014]

[0009] The following drawings form a part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.

[0015]

[0010] Figure 1 depicts the anti-epileptic effects of urolithin A on picrotoxin (PTX) induced seizure like activity (SLA) in Drosophila melanogaster larvae; wherein (A) represents still shots taken from continuous video imaging represents images with the positions of the larvae in different treatment groups, on a time scale of 0-5 seconds; (B) representative confocal images showing increased branching (divergent points on synaptic arbour) are marked with arrowheads. As a compensatory mechanism to increased SLA, the secondary branches increased in PTX, but reduced in number as well as length in PTX+UA (marked in peach) group; (C) represents increased evoked Excitatory Junction Potentials (EJPs) in PTX (marked in red) whereas, urolithin A reduces the evoked EJP marked as an amplitude; (D) depicts increased amplitude of miniature transmission in PTX group (marked in red) is reduced with Urolithin- A (marked in peach). Note that Urolithin- A had no effects on the frequency of miniature transmission (data not shown). Significance was determined using ANOVA test to compare the mean values ± SEM, in accordance with an embodiment of the present disclosure.

[0016] [Oil] Figure 2 depicts the chronic effects of urolithin A on homeostatic modifications of synapses, determined using bang senseless mutant, bss1Drosophila (model of drug-resistant Epilepsy); wherein (A) is a schematic representation showing that Urolithin-A increased mean recovery time in response to electric shock because of decrease in neuronal firing of provoked seizures; (B) is a graphical representation showing that Urolithin-A increased mean recovery time in response to electric shock because of decrease in neuronal firing of provoked seizures; (C) depicts that Urolithin-A potentiates synaptic transmission in both healthy, as well as epileptic context, by an increase in branching and total number of boutons; (D) & (E) depicts that Urolithin-A decreased evoked EJP in the epileptic context but displayed no effects on the miniature synaptic transmission.

[0017]

[0012] Figure 3 depicts the anti-epileptic effects of urolithin A on pentylenetetrazol (PTZ)-induced SLA in the mice, wherein (A) is a schematic representation of the treatment regimen; (B) depicts dose-dependent effects of urolithin A (50 mg / Kg and 100 mg / Kg) in the PTZ chronic kindling model; (C) depicts the Racine scale for seizure development and severity used in Racine score analysis; (D) & (E) depicts traces for basal synaptic transmission, Urolithin A ameliorated the PTZ-induced reduction in basal synaptic transmission. 3 / 7 mice were found to be seizure free during the whole treatment regimen (data not shown). Note the increase in amplitude and slope mediated by urolithin-A (peach colour); (F) depicts Urolithin A increased the spontaneous inhibitory transmission at an amplitude level; (G) represents quantitation for spontaneous IPSC amplitude data points. Significance was determined using ANOVA test to compare the mean values ± SEM, in accordance with an embodiment of the present disclosure.

[0018]

[0013] Figure 4 depicts that Urolithin-A exerts no acute or symptomatic effects on 4AP-0Mg2+ induced epileptiform firing; A) Regular firing subicular neurons showing voltage response to a step depolarization of 200 pA for 500 ms (currentclamp protocol shown with voltage response). B) Representative recording of voltage response of a subicular neuron in whole-cell current-clamp gap-free mode when perfused with 4AP-0Mg, and in the presence of UA (50 pm). C) Instantaneous spike frequency vs time plotted from representative traces shown in B. D) Summary plot of mean spike frequency showing no change after UA application. n=number of slices. n=3. *p < 0.5, **p < 0.05, ***p < 0.001, two-tailed t-test. Error bars represent SEM. DESCRIPTION OF THE INVENTION

[0019]

[0014] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features.

[0020] Definitions

[0021]

[0015] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These definitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.

[0022]

[0016] The articles “a”, “an” and “the” are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

[0023]

[0017] Throughout this specification, unless the context requires otherwise the word “comprise”, and variations such as “comprises” and “comprising” are used in the inclusive, open sense, and will be understood to imply the inclusion of a stated element or step or group of element or steps but not the exclusion of any other element or step or group of element or steps. It is not intended to be construed as “consists of only”.

[0024]

[0018] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably.

[0025]

[0019] The term “seizure”, as used herein, refers to sudden, uncontrolled electrical disturbance in the brain, which cause changes in movement, sensation, awareness, thought or behavior, depending on the part of the brain the seizure affects. Types of seizure include, but is not limited to, absence seizures, atonic seizures, atypical absence seizures, clonic seizures, epileptic or infantile spasms, febrile seizures, focal bilateral tonic clonic seizures (secondarily generalized seizures), focal onset aware seizures (simple partial seizures), focal onset impaired awareness seizures (complex partial seizures), gelastic and dacrystic seizures, myoclonic seizures, drug-resistant seizures, new terms seizure classification, tonic-clonic seizures, or tonic seizures.

[0026]

[0020] The term “epilepsy syndrome”, as used herein, refers to a disorder / disease of the central nervous system and is defined by a cluster of clinical features of a patient’s seizures. The terms “epilepsy syndrome” and “epilepsy” are used interchangeably herein. Examples of epilepsy syndrome include, but are not limited to, Frontal lobe epilepsy, infantile spasms, febrile seizures, progressive myoclonus epilepsy of Lafora, Generalized Epilepsy with Febrile Seizures (GEFS+), Severe Myoclonic Epilepsy of Infancy (SMEI), Benign Neonatal Familial Convulsions (BFNC), Ohtahara Syndrome, early myoclonic encephalopathies, migrating partial epilepsy, infantile epileptic encephalopathies, Tuberous Sclerosis Complex (TSC), focal cortical dysplasia, Type I Lissencephaly, Miller-Dieker Syndrome, Angelman's syndrome, Fragile X syndrome, epilepsy in autism spectrum disorders, subcortical band heterotopia, Walker- Warburg syndrome, posttraumatic epilepsy, progressive myoclonus epilepsies, reflex epilepsy, Rasmussen's syndrome, temporal lobe epilepsy, limbic epilepsy, status epilepticus, abdominal epilepsy, massive bilateral myoclonus, catamenial epilepsy, Jacksonian seizure disorder, Alzheimer’s disease, Unverricht- Lundborg disease, or photosensitive epilepsy, Dravet syndrome, idiopathic generalized epilepsy, juvenile absence epilepsy, juvenile myoclonic epilepsy, Landau Kleffner syndrome, Lennox Gastaut syndrome, myoclonic astatic epilepsy, childhood absence epilepsy, west syndrome, Panayiotopoulos syndrome, benign rolandic epilepsy, refractory epilepsy, or childhood idiopathic occipital epilepsy.

[0027]

[0021] The term “anti-epileptic drug”, as used herein, refers to drugs that are used for treating epilepsy. Examples of anti-epileptic drugs include, but are not limited to, acetazolamide, benzodiazepine, cannabadiols, carbamazepine, clobazam, clonazepam, eslicarbazepine acetate, ethosuximide, ethotoin, felbamate, fenfluramine, fosphenytoin, gabapentin, ganaxolone, huperzine A, lacosamide, lamotrigine, levetiracetam, nitrazepam, oxcarbazepine, perampanel, piracetam, phenobarbital, phenytoin, potassium bromide, pregabalin, primidone, retigabine, rufinamide, valproic acid, sodium valproate, stiripentol, tiagabine, topiramate, vigabatrin, or zonisamide. In an exemplary embodiment, the AED is valproic acid, sodium valproate, clonazepam, ethosuximide, felbamate, gabapentin, carbamazepine, oxcarbazepine, lamotrigine, levetiracetam, benzodiazepine, phenobarbital, pregabalin, primidone, tiagabine, topiramate, potassium bromide, phenytoin, stiripentol, vigabatrin, or zonisamide. In an exemplary embodiment, the AED is valproic acid, sodium valproate, gabapentin, topiramate, carbamazepine, oxcarbazepine, or vigabatrin.

[0028]

[0022] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.

[0029]

[0023] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.

[0030]

[0024] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein.

[0031]

[0025] Embodiments of the present disclosure provides a method for the treatment of epilepsy syndrome and / or seizure in a subject. The method disclosed herein comprises administering a therapeutically effective amount of a compound, or salt thereof. Embodiments herein specifically target epileptic mediators. Embodiments herein further overcome the challenges that are generally observed with the use of existing anti-epileptic drugs, such as poor patient response, severe physical and / or psychological side effects and prevalence of drug resistance, etc. Also, embodiments herein are capable of mitigating the behavioral, synaptic (structural and functional) as well as molecular defects of excitatory / inhibitory imbalances in subjects. Further, the present disclosure discloses a compound for use in treating epilepsy syndrome and / or seizure. Also disclosed herein is a composition for the treatment of epilepsy syndrome and / or seizure.

[0032] Method

[0033]

[0026] In an embodiment of the present disclosure, there is provided a method for the treatment of epilepsy syndrome and / or seizure in a subject, comprising administering a therapeutically effective amount of a compound, or salt thereof, of Formula I,

[0034] Formula I wherein A, B, C, X, Y, and Z are independently selected from H or OH groups.

[0035]

[0027] The compound, in an embodiment, is having a formula of Formula I wherein one or more of A, B, C, X, Y, and Z are, independently, H groups. In another embodiment, the compound is of Formula I wherein one or more of A, B, C, X, Y, and Z are, independently, OH groups. In an embodiment of the present disclosure there is provided a method as described herein, wherein said compound is at least one selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E. In a preferred embodiment, the compound is urolithin A.

[0036]

[0028] The salt of compound of Formula I, according to embodiments herein, may be a pharmaceutically acceptable salt of the compound of Formula I. In an embodiment, the salt is a salt of an acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methane sulfonic acid, ethane sulfonic acid, p-toluene sulfonic acid, salicylic acid, or combination thereof.

[0037]

[0029] In another embodiment the salt is selected from sodium salt, potassium salt, lithium salt, ammonium salt, calcium salt, magnesium salt, iron salt, zinc salt, copper salt, manganese salt, aluminium salt, or combination thereof.

[0038]

[0030] The compound of Formula I, according to embodiments herein, is commercially available through various vendors. The compound may be procured from any such suitable vendors. Alternatively, the compound may be prepared by methods generally known in the art. It is understood that the various processes, reactants and intermediates using which the compound of Formula I can be synthesized, would be apparent to a person skilled in the art. Any such suitable processes, reactants and intermediates may be used in order to obtain the compound, according to embodiments herein.

[0039]

[0031] Embodiments herein may be used in treatment of any epilepsy syndrome. In an example, the syndrome is selected from Dravet syndrome, idiopathic generalized epilepsy, juvenile absence epilepsy, juvenile myoclonic epilepsy, Landau Kleffner syndrome, Lennox Gastaut syndrome, myoclonic astatic epilepsy, childhood absence epilepsy, west syndrome, Panayiotopoulos syndrome, benign rolandic epilepsy, refractory epilepsy, or childhood idiopathic occipital epilepsy.

[0040]

[0032] Further, embodiments herein are capable of treating seizures of epilepsy. In an example, the seizure is selected from simple focal seizure, complex focal seizure, secondary generalized seizure, absence seizure, or tonic clonic seizure.

[0041]

[0033] It is understood that the nature and frequency of the seizures may vary from person to person. Based on the type of seizures, age of onset, causes of seizures, severity of seizures, electrographic patterns, etc., epilepsy may be categorized into various epilepsy syndromes. Embodiments herein can treat all such seizures and / or syndromes in a subject. The term “subject”, as used herein, includes a mammal, particularly human, in need of such treatment or having / suspected of having epilepsy syndrome and / or seizure.

[0042]

[0034] Further, the mode of administration, treatment regimen, dosage strength, dosage form etc. may vary depending on factors such as the age of the subject, nature of disease, severity of seizures, etc. Accordingly, the dosage form, mode of administration, treatment regimen, etc. may suitably be tailored based on such factors. The compound of Formula I is administered in a therapeutically effective amount. In an embodiment, the therapeutically effective amount of the compound is in the range of 4 to 25 mg / kg. In another embodiment, the therapeutically effective amount of the compound is ranging from 5 to 25 mg / kg, 10 to 25mg / kg, 15 to 25 mg / kg or 20 to 25 mg / kg.

[0043]

[0035] In an embodiment, the method comprises administering at least one compound selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E, or salt thereof. In another embodiment, the method comprises administering a salt of at least one compound selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E, wherein the salt is a salt of an acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methane sulfonic acid, ethane sulfonic acid, p-toluene sulfonic acid, salicylic acid, or combination thereof; or a salt selected from sodium salt, potassium salt, lithium salt, ammonium salt, calcium salt, magnesium salt, iron salt, zinc salt, copper salt, manganese salt, aluminium salt, or combination thereof.

[0044]

[0036] Further, embodiments herein provide a compound for use in treatment of epilepsy syndrome and / or seizure in a subject.

[0037] In an embodiment, the compound is a compound, or salts thereof, of Formula

[0045] I

[0046] Formula I wherein A, B, C, X, Y, and Z are independently selected from H or OH groups, for use in the treatment of epilepsy syndrome and / or seizure in a subject.

[0047]

[0038] A, B, C, X, Y, and Z, in Formula I, may suitably be selected from H or OH groups. In an embodiment, said B and X independently are OH groups, and said A, C, Y and Z independently are H groups. In another embodiment, said B is OH groups, and said A, C, X, Y and Z independently are H groups. Further, in another embodiment, said B, X and Y independently are OH groups, and said A, C and Z independently are H groups. In an embodiment, said B, A, X and Y independently are OH groups, and said C and Z independently are H groups. In yet another embodiment, said B, X, Z and A independently are OH groups, and said A and C independently are H groups.

[0048]

[0039] Accordingly, in an embodiment, the compound is at least one selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E.

[0049]

[0040] In an embodiment of the present disclosure there is provided a compound as described herein for use in the preparation of a medicament for the treatment of epilepsy syndrome and / or seizure in a subject.

[0050] Composition

[0051]

[0041] Embodiments herein disclose a composition for use in the treatment of epilepsy syndrome and / or seizure in a subject.

[0042] In an embodiment, the composition comprises a compound, or salts thereof, of Formula I

[0052] Formula I wherein A, B, C, X, Y, and Z are independently selected from H or OH groups; and at least one pharmaceutically acceptable carrier, for use in the treatment of epilepsy syndrome and / or seizure in a subject.

[0053]

[0043] In an embodiment, the composition comprises at least one compound of Formula I wherein the B and X independently are OH groups and the A, C, Y and Z independently are H groups; or B is OH groups, and the A, C, X, Y and Z independently are H groups; the B, X and Y independently are OH groups, and the A, C and Z independently are H groups; the B, A, X and Y independently are OH groups, and the C and Z independently are H groups; the B, X, Z and A independently are OH groups, and the A and C independently are H groups. In an embodiment, the composition comprises at least one compound selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E.

[0044] In a preferable embodiment, the compound is urolithin A.

[0054]

[0045] The composition, according to embodiments herein, may further include one or more anti-epileptic drug. Accordingly, embodiments herein may be in the form of a combination therapy. In an embodiment, the composition further comprises at least one anti-epileptic drug selected from clobazam, gabapentin, primidone, rufinamide, stiripentol, vigabatrin, or combinations thereof.

[0046] The composition, in embodiments herein, may comprise the compound in therapeutically effective amounts. The therapeutically effective amount, according to embodiments herein, would depend on factors such as the age of the subject, nature of disease, severity of seizures, etc. In an embodiment, the composition comprises the compound in a therapeutically effective amount ranging from 4 to 25 mg / kg with respect to the composition. In another embodiment, the composition comprises the compound in an amount ranging from 5 to 25 mg / kg, 10 to 25mg / kg, 15 to 25 mg / kg, or 20 to 25 mg / kg with respect to the composition. Further, the amount of anti-epileptic drug may also be varied in various embodiment.

[0055]

[0047] The composition, according to embodiments herein, further comprises pharmaceutically acceptable carriers. In an embodiment, the pharmaceutically acceptable carrier is selected from adjuvant, preservative, excipient, diluent, or combinations thereof. Various pharmaceutically acceptable carriers are conventionally known for use in pharmaceutical compositions. Embodiments herein may include any such suitable pharmaceutically acceptable carriers. Examples of suitable pharmaceutically acceptable carriers include lactose, mannitol, sucrose, corn starch, potato starch, crystalline cellulose, cellulose derivatives, gelatin, sodium carboxymethylcellulose, carboxymethylcellulose, glucose, dimethyl sulfoxide, water, etc. Alternatively, carriers may also include liposomal carriers formed using natural or synthetic lipids including, but not limited to, polyethylene glycol (PEG), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), and lecithin; biocompatible nanoparticles for eg: poly (lactic-co-glycolic acid) nanoparticles, starch-based nanoparticles, chitosan / alginate nanoparticles, nanoemulsions, etc. Accordingly, in an embodiment, the pharmaceutically acceptable carrier is selected from lactose, mannitol, sucrose, com starch, potato starch, crystalline cellulose, cellulose derivatives, gelatin, sodium carboxymethylcellulose, carboxymethylcellulose, glucose, dimethyl sulfoxide, water, polyethylene glycol (PEG), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), lecithin, poly (lactic-co-glycolic acid) nanoparticles, starch-based nanoparticles, chitosan / alginate nanoparticles, or combination thereof.

[0056]

[0048] In an embodiment of the present disclosure, there is provided a composition comprising said compound as described herein and at least one pharmaceutically acceptable carrier, for use in the treatment of epilepsy syndrome and / or seizure in a subject selected from adjuvant, preservative, excipient, diluent, or combinations thereof; further comprises at least one anti-epileptic drug selected from clobazam, gabapentin, primidone, rufinamide, stiripentol, vigabatrin, or combinations thereof; and said compound is at least one selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E.

[0057]

[0049] The composition, according to embodiments herein, may be prepared by methods generally known in the art. It is understood that the various processes and carriers would be apparent to a person skilled in the art. Any such suitable processes, and carriers may be used to obtain the composition, according to embodiments herein.

[0058]

[0050] In an embodiment, the method for preparation of the composition comprises adding Urolithin-A to a solution of Dimethyl sulfoxide (DMSO), preferably of 0.5 wt% DMSO, and vortexing to achieve complete solvation of Urolithin-A.

[0059]

[0051] In an embodiment, the method for preparation of the composition comprises adding Urolithin-A to a solution of carboxy methyl cellulose (CMC), preferably of 0.5 wt% CMC, and vortexing to achieve complete solvation of Urolithin-A.

[0060]

[0052] In an exemplary embodiment, the composition is a pharmaceutical composition as depicted herein below:

[0061] Table 1:

[0053] The composition, according to embodiments herein, may be formulated into any suitable dosage form including but not limited to, solid, liquid, semi-solid or gaseous, for example tablet, capsule, powder, granules, solution, suspension, injectable, inhalant, gels, etc. Administration of the composition or compound, according to embodiments herein, may be achieved in various ways including, but not limited to, oral or parenteral, buccal, dermal, transdermal, intracheal, etc. In a preferred embodiment, the composition is a tablet for oral administration.

[0062] EXAMPLES

[0063]

[0054] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices, and materials are described herein. It is to be understood that this disclosure is not limited to methods, and experimental conditions described, as such methods and conditions may apply.

[0064] Materials

[0065]

[0055] Urolithin A (UA) was procured from Hefei Hirisun Pharmatech Ltd., Picrotoxin and Pentylenetetrazol (PTZ) was procured from Sigma Aldrich.

[0066] Example 1

[0067] Determination of effects of urolithin A on the compensatory synaptic mechanisms of epilepsy

[0068]

[0056] Sample preparation: 22.8 pg (100 pM) of urolithin-A was dissolved in 0.5% v / v Dimethyl Sulphoxide and mixed with 1ml of Drosophila food in each vial.

[0069]

[0057] Procedure: The effects of urolithin A on the compensatory synaptic mechanisms of epilepsy were determined by testing urolithin A in a non-acquired model of epilepsy, such as picrotoxin (PTX) induced seizures in Drosophila melanogaster larvae (procured from Indian Institute of Science), at the time point of 4 hours of co-feeding. For this study, Urolithin-A dissolved in 0.5% Dimethyl sulfoxide (DMSO) was used. The phenotype was scored at behavioral, synaptic structural as well as functional levels (Figure 1). Administration of Urolithin-A improves the defects in the locomotory contractions of PTX fed larvae (Figure 1A). PTX induces increased synaptic branching as an acute compensatory mechanism to increased neuronal firing, which is observed as the divergent points of the synaptic arbor with ghost boutons, marked with arrows (Figure IB). Urolithin A resulted in the reduction of the secondary branching in the PTX-UA group. Further urolithin A also resulted in the reduction of evoked Excitatory Junction Potentials (EJPs) and the amplitude of miniature transmission (Figure 1C-D).

[0070]

[0058] The chronic effects of urolithin A on homeostatic modifications of synapses were determined using bang senseless mutant, bss1Drosophila (model of drugresistant Epilepsy (Figure 2). bss1represents Gain of Function (GOF) in the sodium channel, the only Drosophila ortholog of human sodium channel. More than 500 variants in the sodium channel have been linked with human Epilepsy. Urolithin-A increased mean recovery time in response to electric shock (Figure 2 A-B) because of decrease in neuronal firing of provoked seizures. As a result of chronic compensatory effects, bss1mutant shows decrease in synaptic arbor with reduction in branches and total number of boutons. Urolithin-A potentiates synaptic transmission in both healthy, as well as epileptic context, by an increase in branching and total number of boutons (Figure 2 C). Urolithin-A decreased evoked EJP in the epileptic context but displayed no effects on the miniature synaptic transmission (Figure 2 D-E).

[0071] Example 2

[0072] Anti-epileptic effects of urolithin A on pentylenetetrazol (PTZ) induced seizure like activity in chronic kindling model of mice

[0073]

[0059] Sample preparation: Urolithin-A (100 mg / Kg) dissolved in 0.5% carboxy methyl cellulose (CMC) was used.

[0060] Procedure: To validate the potential anti-epileptic effects mediated by urolithin A in the epileptic models of Drosophila, urolithin A was further tested in PTZ induced chronic kindling model of mice. For this study, Urolithin-A dissolved in 0.5% carboxy methyl cellulose (CMC) was used. Effects were scored at the behavioral level (Racine score analysis), as well as synaptic transmission at the network and single cell level. To generate the kindling epilepsy model, mice (procured from Jawaharlal Nehru Centre for Advanced Scientific Research) were intraperitoneally injected with PTZ (35 mg / kg) once every other day for at least eleven total injections, and mice showing more than three consecutive stage 4 seizures were fully kindled. Vehicle control animals were injected with saline. Seizure events during a 30 min period after each PTZ injection were observed. Urolithin A was administered 60 minutes prior to the dose of PTZ (Figure 3 A). The seizure intensity was scored as follows: Stage 0, no response; Stage 1, behavioural arrest; Stage 2, tail straightening; Stage 3, myoclonic jerks, and rearing; Stage 4, turning over the sides with myoclonic and tonic-clonic jerks; Stage 5, generalized tonic-clonic seizures with wild jumping; and Stage 6, death (Figure 3B). Mice were sacrificed after the 11thinjection, and hippocampal slices were prepared for basal synaptic transmission studies and patch-clamping.

[0074]

[0061] Urolithin A consistently resulted in lower seizure scores over the twenty -two days of study when compared to non-urolithin treated PTZ-induced mouse models (Figure 3B). Urolithin A ameliorated the PTZ-induced reduction in basal synaptic transmission (Figure 3 D-E) and increased the spontaneous inhibitory transmission at an amplitude level (Figure 3 F-G). Urolithin A shows no effects on the frequency of neither excitatory, nor inhibitory transmission at spontaneous and miniature levels indicating a post-synaptic effect of Urolithin-A.

[0075] Example 3

[0076] Effects of Urolithin-A 4AP-0Mg2+ induced epileptiform firing

[0077]

[0062] Sample preparation: 11.4 pg (50 pM) of urolithin-A was dissolved in 0.5% v / v Dimethyl Sulphoxide and mixed with the required volume of artificial Cerebrospinal fluid (aCSF).

[0063] Procedure: Anti-epileptic potential of Urolithin-A at the acute level (seconds / minutes) by infusing the 4 Aminopyridine-0 magnesium (4AP-0Mg) induced epileptiform slices with Urolithin-A (50 pM). Firstly, the subicular neurons were characterized by measuring the voltage traces in response to the current protocol in a normal ACSF solution (Fig. 4a). For inducing epileptiform activity, 4AP+Magnesium free ACSF (4AP-0Mg2+) was used. Epileptiform activity was measured by quantifying the instantaneous spike frequency after 4AP-0Mg2+ induction. After induction of epileptiform activity, slices were perfused with Urolithin-A (50 pM) in the presence of 4AP-0Mg2+. A range of classical symptomatic treatments with the potency of the seizure- spread blockade has shown seizure- suppressive effects in the 4AP-0Mg model; however, UA (50 pM) didn’t show a reduction in the frequency of the spikes, as illustrated in the representative traces and summary plots.

[0078]

[0064] Results: Figure 4 depicts that Urolithin-A exerts no acute or symptomatic effects on 4AP-0Mg2+ induced epileptiform firing; A) Regular firing subicular neurons showing voltage response to a step depolarization of 200 pA for 500 ms (current-clamp protocol shown with voltage response). B) Representative recording of voltage response of a subicular neuron in whole-cell current-clamp gap-free mode when perfused with 4AP-0Mg, and in the presence of UA (50 pm). C) Instantaneous spike frequency vs time plotted from representative traces shown in B. D) Summary plot of mean spike frequency showing no change after UA application. n=number of slices. n=3. *p < 0.5, **p < 0.05, ***p < 0.001, two-tailed t-test. Error bars represent SEM.

[0079] Advantages of the present disclosure

[0080]

[0065] The present disclosure provides a method for the treatment of epilepsy syndrome and / or seizure in a subject, comprising administering a therapeutically effective amount of the compound or salt thereof as disclosed herein.

[0081]

[0066] The compound exhibits anti-epileptic effect by mitigating the behavioral, synaptic (structural and functional) as well as molecular defects of excitatory / inhibitory imbalances, as observed in drosophila and mouse models of epilepsy. Therefore, the compound is effective in targeting epileptic mediators and thereby chronic epilepsy.

Claims

I / We claim:

1. A method for the treatment of epilepsy syndrome and / or seizure in a subject, comprising administering a therapeutically effective amount of a compound, or salt thereof, of Formula IFormula I wherein A, B, C, X, Y, and Z are independently selected from H or OH groups.

2. The method as claimed in claim 1, wherein said compound is at least one selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E.

3. The method as claimed in claim 1, wherein said salt is a salt of an acid selected from hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methane sulfonic acid, ethane sulfonic acid, p-toluene sulfonic acid, salicylic acid, or combination thereof.

4. The method as claimed in claim 1, wherein said salt is selected from sodium salt, potassium salt, lithium salt, ammonium salt, calcium salt, magnesium salt, iron salt, zinc salt, copper salt, manganese salt, aluminium salt, or combination thereof.

5. The method as claimed in claim 1, wherein said epilepsy syndrome is selected from Dravet syndrome, idiopathic generalised epilepsy, juvenile absence epilepsy, juvenile myoclonic epilepsy, Landau Kleffner syndrome, Lennox Gastaut syndrome, myoclonic astatic epilepsy, childhood absence epilepsy, west syndrome, Panayiotopoulos syndrome, benign rolandic epilepsy, refractory epilepsy, or childhood idiopathic occipital epilepsy.

6. The method as claimed in claim 1, wherein said seizure is selected from simple focal seizure, complex focal seizure, secondary generalized seizure, absence seizure, or tonic clonic seizure.

7. The method as claimed in claim 1, wherein the therapeutically effective amount of the compound is in the range of 4 to 25 mg / kg.

8. A compound, or salts thereof, of Formula Iwherein A, B, C, X, Y, and Z are independently selected from H or OH groups, for use in the treatment of epilepsy syndrome and / or seizure in a subject.

9. The compound as claimed in claim 8, wherein said compound is at least one selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E.

10. The compound as claimed in claim 8, for use in the preparation of a medicament for the treatment of epilepsy syndrome and / or seizure in a subject.

11. A composition comprising a compound, or salts thereof, of Formula IFormula I wherein A, B, C, X, Y, and Z are independently selected from H or OH groups; and at least one pharmaceutically acceptable carrier, for use in the treatment of epilepsy syndrome and / or seizure in a subject.

12. The composition as claimed in claim 11, wherein said compound is at least one selected from the group consisting of urolithin A, urolithin B, urolithin C, urolithin D and urolithin E.

13. The composition as claimed in claim 11 , further comprising at least one antiepileptic drug selected from clobazam, gabapentin, primidone, rufinamide, stiripentol, vigabatrin, or combinations thereof.

14. The composition as claimed in claim 11, wherein said compound is in a therapeutically effective amount in the range of 4 to 25 mg / kg with respect to the composition.

15. The composition as claimed in claims 11, wherein said pharmaceutically acceptable carrier is selected from adjuvant, preservative, excipient, diluent, or combinations thereof.

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