WO2017060916A1

WO2017060916A1 - Metallic nanoparticle alone and/or in combination as novel agent for the treatment of uncontrolled electric conductance related disorders and/or seizure, epilepsy & convulsions.

Info

Publication number: WO2017060916A1
Application number: PCT/IN2016/000245
Authority: WO
Inventors: NOUSHAD Javed Md; SABIR Alam Md; Hyder Pottoo Faheem
Original assignee: NOUSHAD Javed Md; SABIR Alam Md; Hyder Pottoo Faheem
Priority date: 2015-10-09
Filing date: 2016-10-10
Publication date: 2017-04-13

Abstract

The present invention relates to pharmaceutical composition/s comprising of at-least one of metallic nanoparticles either alone or in their combinations, more preferably such metallic nanoparticles here may be of gold, silver and zinc nanoparticles; more particularly preferred if such composition may also contain at-least one of drugs from the category such as antiseizure, antiepileptic, antioxidants or nutraceuticals or other therapeutic class having potential to be use in such disorders, so that these compositions would be useful in therapeutic or prophylactic treatment as well as the management of pathophysiological or pathological states, arising either due to unregulated or unbalanced or dysregulated electrical conductance; such as (but not limited to) mainly convulsions and/or seizures and/or epilepsies etc. The present invention also discloses a more preferable synergistic and potentiating pharmaceutical composition; which may be combination of at-least one of metallic nanoparticles with the carbamazepine; more specifically to say such preferred metallic nanoparticles here may be gold, silver and zinc nanoparticles; as therapeutic or prophylactic treatment for the management of any pathophysiological or pathological states arising due to unregulated or unbalanced or dysregulated type of electric conductance disorders preferably such as convulsions, seizures or epilepsies.

Description

DISCRETION

TITLE OF INVENTION:

Metallic nanoparticle alone and/or in combination as novel agent for the treatment of uncontrolled electric conductance related disorders and/or seizure, epilepsy & convulsions.

OBJECTIVES OF THE INVENTION:

The current anti-epileptic drugs provide symptomatic relief in epilepsy while the underlying pathology remains largely unabated, hence the objective of current invention was to search for safe and effective novel agent(s) with a potential to modulate and/or control and/or regulate uncontrolled and/or unbalanced electrical flow and/or maintain electric current at an ionic path by using metallic nanoparticles which may be used either alone or in combination or in conjugation with or one or more of anti-convulsant and/or anti-epileptic/s and/or antioxidants and/or nutraceuticals and/or other therapeutic agent/s and/or pharmaceutically acceptable excipient/s, hence may be used as novel therapeutics agent/s and/or drugs for the treatment of and/or to manage and/or to regulate the pathophysiological/pathological conditions which are supposed to be due to uncontrolled nature of electrical conductance activity or in conditions which are expected to arise due to the uncontrolled electrical conductance and/or similar to or expected to lead the physiological state and/or seizures and/or epileptic seizures and/or convulsions etc.

DESCRIPTION OF INVENTION:

Process involve clinical significance of metallic nanoparticles formulation for their usage either alone or when they are present in the form of conjugates or in combination with one or more anticonvulsant and/or anti-epileptics and/or other therapeutic agents and/or excipients which are pharmaceutically acceptable, in the pathophysiological conditions in which electric conductance activity is uncontrolled or unregulated or unbalanced and/or conditions which are similar to these state and/or conditions which are expected to lead/rise because of the above state.

Hence potential of administering these formulations as well as their clinical significance in, as part of therapy or co-therapy in the treatment of and/or for the management of and/or for the regulation of any pathophysiological conditions similar to as mentioned in the above paragraph. We propose here that metallic nanoparticles alone, or in synergistic combination with anti- epileptic drugs and/or anticonvulsant drugs and/or antioxidants and/or nutraceuticals interfere with the propagation of abnormal electric signal conductance which may be useful in the treatment of pathophysiological state which arises under such circumstances,

DETAILED DISCRIPTION

The following detailed description of the invention is provided here only for better in understanding of the invention as well as concept. Hence, No unnecessary limitations are to be understood therefrom. To those skilled in the art, modifications will become obvious upon reading the disclosure and may be made without departing from the spirit of the invention and scope of the appended claims. So, all such equivalents to the foregoing aspects will now be readily apparent to the person skilled in the art.

For example, here, for reducing agent, we preferably select reducing sugar and according to one of the embodiment or aspect here of the invention, was preferably sourced from gum ghatti (Anogeissus latifolia. India), but other reducing agent/s or reducing sugar/s or their combinations may be apparently selected by any person known as the person skilled in the art, hence may also be substituted therefor.

Also, For example, here, according to an embodiment of the invention; microwave was used as preferable source of energy to provide a reducing environment, however another source of energy such as ultraviolet light, sunlight, direct heating, biometabolic energy etc., may also be substituted therefor by any person known such art.

STEP 1: Preparation of Gum powder for use in the formulation Optimized Formula 1 : For preparation of raw gum solution

Gum ghatti : 10 gm Water: 100 mL Temperature: 60 °C Duration: 4 hours In the below mentioned process we first process to isolate water soluble fraction of gum solution by filtration of insoluble residue if any, followed by water soluble fraction solution processed as lyophilized powder which were readily available to us during the time of preparation of formulation, hence at the time of formulation appropriately weigh and diluted there.

Process for lyophilizing powder involved lyophilized for upto 72 hours. However further addition of cryoprotectant (1-10 %) was not essentially required but we found that their addition enhances the stability as well as fasten the process only if used in optimum ratio, as well as such process also an aid in the final products too. Lyophilized Powder were prepared, collected and may be dispensed or formulated in appropriate dosage form as per suitability.

STEP 2 : Preparation of metallic nanoparticles

In the below mention process, we found that double distilled water sufficient to give stable formulation however here, we intentionally avoid the buffer systems for specific reasons however buffer system of suitable pH may be also be substituted thereof too. Generic Formula 1: Gold nanoparticles synthesis by heating method

Optimized Formula code: GN

Gold solution (strength 0.1 Molar) : 40

Gum ghatti solution (1% solution of lyophilized powder) : 4 ml Water (Quantity sufficient) : 10 mL Heating Time: 5 min

Heating Temperature: 80° C

Generic formula 2 : Gold Nanoparticles synthesis by Microwave method Optimized Formula code: GF

Gold solution (strength 0.1 Molar) : 40 Gum ghatti solution (1% solution of lyophilized powder) : 4 ml Water (Quantity sufficient) : 10 mL Microwave Time: 30 Sec

Generic Formula 3: Silver nanoparticles synthesis by Microwave method Optimized Formula code: SF Silver solution (0.1 Molar Silver Nitrate) : 20 μΐ.

Gum ghatti solution (1% solution of lyophilized powder) : 4 ml

Water (Quantity sufficient) : 10 mL

Microwave Time: 30 Sec

Microwave power: 700 watt Generic Formula 4: Zinc nanoparticles synthesis by Microwave method

Optimized Formula code: ZF

Zinc solution (0.1 Molar Zinc Nitrate) : 20 μΐ-

Gum ghatti solution (1% solution of lyophilized powder) : 4 ml

Water (Quantity sufficient) : 10 mL Microwave Time: 30 Sec

Microwave power: 700 watt

In brief; for heating process, lyophilized powder is mixed with required quantity of water under constant predetermined stirring followed by heating till desired temperature are reached and it was maintained throughout the experiment, followed by addition of metallic solution upto desired time. Changing of colour indicates synthesis of metallic nanoparticles.

While, for Microwave process, lyophilized powder is mixed with required quantity of water under constant predetermined stirring, followed by addition of metallic solution and microwave radiation are used there of above mentioned strength upto desired time. Changing of colour indicates synthesis of metallic nanoparticles.

STEP 3: Loading of drug to formulation The drugs as mentioned in the claim 2 here, may be loaded in various ways for preparation of combined dosage form from the above prepared metallic nanoparticle formulation. Way 1 example: Optimized formula Code L-1

Gold nanoparticles solution: 100 μL Carbamazepine: 10 mG

Both are incubated together overnight. % of drug loading can be analysed by removing free drug part after processing of centrifugation. This art is well known so need not to further described much in details here.

Way 2 example: Optimized formula Code L-2

Gold nanoparticles solution: 100

Carbamazepine: 10 mG

Lyophilizing agent: Mannitol (2%) Both are incubated together overmght. % of drug loading can be analysed by removing free drug part after processiang of centrifugation. This art is well known so need not to further described much in details here. The drug loaded solution further lyophilized using suitable any cryoprotectant agent to prepare lyophilized powder, preferably stored at 4°C,

Way 3 example: Optimized formula Code L-3 Gold nanoparticles solution: 100 μL· Carbamazepine: 10 mG

Both are stored in two isolated containers in fixed amount at required temperature, more preferably 4°C, however both are now ready to be mixed just before the dosing.

Way 4 example: Optimized formula Code L-4 Gold nanoparticles powder: equivalent to 100 μL. gold solution Carbamazepine: 10 mG

Both are in the powder forms and to be mixed together with water before dosing. Further Notes:

Here in above example water may also be substituted with other solvents or appropriate buffer system as per need.

The all above prepared formulations may be immediately stored at 4°C or better to immediately lyophilize or spray dried or using others well known arts to prepare in dry powder form.

Formula 1: For Capsules Optimized formula Code C-1

For the capsule preparation, we may take either liquid formulation or lyophilized form of previous prepared composition as per suitability or need - Lyophilized powder: 10 mg

HPMC: 100 mg

Capsule type: Hard gelatine

Such powder preparation may be filled in a hard gelatine capsules and stored at cool and dry place, better if stored at 4 °C. Formula 2: For Tablet

Optimized formula Code T-1 and Lyophilized powder: 10 mg HPMC: 100 mg

Method: direct compression punch method For the Tablet preparation, we may take final composition as mentioned above (in Formula 1 : For Capsules) followed by compression under the punch machine. Method can be optimized further as well as other pharmaceutical active agents such as disintegrating agent, compressing agent, binders or diluents etc., may be further incorporated as per suitability and if need arises thereof, These powder may be filled in capsules or formulated as tablets either alone or in presence of pharmaceutically acceptable excipients, such arts are well known and suitable process may be accepted here too. Hence here, we here don't want to discuss on that anymore, seeming not much relevant to current invention.

ANIMAL STUDY

Model Name: MES model in mice Model Used: MES model

Some of investigated Animal Study results of MES model in mice showing synergistic efficacy and potentiating effects of few selected compositions.

Effect of formulations on THLE and NO- THLE

Interpretation of above graphs:

As shown in above graphs, the lowest dose of carbamazepine (CB 0 = lmg) as well as metallic nanoparticles (GF 0 = 150 μΐ,; SF 0 = 200 μί, ZF 0 = 100 μί) if used alone, were therapeutic not effective (therapeutic ineffective dose) i.e., show no safety there, but if their combination were used it started to exhibit therapeutic response in term of safety in the mice there. Such types of synergistic effects were also achieved in a composition containing combination of other dose strengths of these such as, CB 1 with GF 1 or SF 1 or ZF 1. From the graph above its clearly shown that if the composition containing combination of Carbamazepine, and the metallic nanoparticles then two components mutually enhance and potentiate the therapeutic effects in synergistic manner.

Further the research established that monotherapy with the increasing dose of gold nanoparticles From GF 0 to upto highest investigated dose here i.e., GF 2 yielded better therapeutic effects , which in case of silver and zinc nanoformulation we found that initially increasing dose of such nanoparticles from SF 0 to SF 1 or ZF 0 to ZF 1 , the therapeutic efficacy were enhanced , in other terms we may also say that at the intermediate investigated dose here which i.e., SF 1 and ZF 1 dose strength therapeutic response of such nanoparticles formulation ,if used alone, were better wrt both highest (SF 2 and ZF 2) and lowest dose strength (SF 0 and ZF 0 ) respectively, in this study

FIELD OF INVENTION:

The present invention relates to the diseases and treatments of the state of uncontrolled electrical conductance activity or the state/condition which arise/lead due to such abnormal activity including such state of central nervous systems which would lead to or expected to lead to seizures and/or epileptic seizures and/or convulsions.

BACKGROUND OF INVENTION:

The neurons communicate with one another by firing tiny electric signals that pass from cell to cell. The activities of brain such as thinking, seeing, feeling, hearing, controlling the movement of muscles, etc. depends upon the precise location of these signals. A seizure triggers when electrical signals in the brain fire in an abnormal and intense manner. However the abnormality can be located in a specific area of brain called partial seizures or throughout the brain called generalized seizures. Rarely, seizures can continue for many minutes or even hours (a serious medical emergency called status epilepticus). A seizure can be provoked by any situation that seriously disturbs the physical or chemical environment of the brain. The current antiepileptic medications include phenobarbital, primidone, phenytoin, carbamazepine, and valproate; as well newer antiepileptic drugs that act by novel mechanisms of voltage-dependent ion channel blockade, enhancement of inhibitory neurotransmission i.e GABA, and/or reduction of excitatory neurotransmission i.e Glutamate. Examples include glutamate antagonism at N-methyl-D- aspartate (NMDA) receptors (e.g. , felbamate) and a- amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMP A) receptors (e.g., felbamate, topiramate) and inhibition of γ-aminobutyric acid (GABA) reuptake in neurons and astrocytes (e.g., tiagabine). Despite the use of these anticonvulsant drugs either alone or in combination, 30% to 40% of patients continue to have seizures (Kwan et al. , N. Engl. J. Med. 2000; 342: 314-319). Epileptic seizures are thus considered as disturbance in the propagation of electrical signal on the path which may be because of abnormality in function of various biological channels. Hence present invention discuss general methods for the synthesis, formulation of gold nanoparticles and their synergistic effect when combined with anti-seizure or anti-convulsant or anti-epileptic drugs. The most important aspect or part of disclosure are clinical usage or therapeutic advantages of said nanoparticles in a way which may be either alone or/and in bimetallic form ; or/and in a way as conjugates or/and their combination with one or more therapeutic agents which may be one or more convulsant and/or antiepileptic agents or else, and/or pharmaceutically acceptable excipients; for the treatment of and/or in the management of and/or in the regulation of pathological state with uncontrolled electrical conductance and/or convulsions and/or seizures and/or epileptic seizures and/or epilepsy and/or related disorders by administering the said nanoparticles to a subject in need thereof. Here we tend to modulate and/or control or/and regulate uncontrolled and/or unbalanced flow and/or maintain electric current at an ionic path by using any external agents, metallic nanoparticles, as said above, in a way as mentioned above so that it would be clinically significant for the treatment of pathophysiological and/or pathological conditions of uncontrolled electrical conduction and/or disorder related to and/or similar to such disorders and/or convulsions/epilepsy and/or seizures using/administering such nanoparticles to the subjects in need thereof.

DISCLOSURE: The present invention discloses here to use of above mentioned surface functionalized and/or encapsulated and/or conjugated form of metallic nanoparticles with/without one or more therapeutic active agents and/or anti-convulsants and/or anti-epileptics and/or anti-seizures and/or pharmaceutical acceptable excipients and/or with semisynthetic and/or natural and/or synthetic compound in a way as mentioned so that it would be used as potential therapeutic agent in the treatment of pathological and/or physiological state of uncontrolled electric conductance activity or state similar to or related to such state and/or epilepsy and/or seizure and/or convulsions etc. .

Here, such method can be adopted to regulate or to treat or to manage the pathological and/or physiological state of uncontrolled electrical conductance and/or conditions, with metal ions which are present either as in nobel form and/or as zero valent metal nanoparticles and/or in oxide forms and/or other stable chemical forms. Such metals can be any one which ranges from as mentioned here ; but not limited to ; e.g., Silver (Ag), Palladium (Pd), Gold (Au) and Iron (Fe) , Zinc (Zn) , Manganese (Mn), Copper (Cu), Platinum (Pt), and Indium or as their bimetals forms; and/or their concentration as low as to 0.01 mM and/or mean particle size upto few μπι; due to and/or related to and/or conditions similar to such pathological and/or physiological state as well as in conditions expected to lead to activity like, but not limited to, seizures and/or epileptic seizures and/or epilepsy and/or related disorders or else in a subject, by administering to a subject metallic nanoparticles, provided synthesized either by any living cells and/or by providing energy and/or on the basis of thermodynamic principles and/or involving any chemical reaction which may be in the presence of sufficient quantity of reducing agent/s sourced from either synthetic or semisynthetic- or plant extract or microorganisms, with or without any solvent and/or surfactant as well as used into the subjects as either alone and/or as bimetallic form and/or in conjugation or in combination with one or more therapeutic agents and/or anti-convulsant and/or anti-epileptic agents and/or pharmaceutical acceptable excipients and/or other agents having properties or potential against such condition .

SUMMARY OF THE INVENTION

There remains a long-felt need for treatments which provide a favourable safety profile and effectively provide relief to the patient suffering from all type of pathological as well as physiological conditions of uncontrolled electric conductance or which are supposed to be attributed by these conditions and/or linked with and/or related to such condition or conditions which arise by such state or lead by such state and/or seizures and/or epileptic seizures and/or convulsions.

Hence the present invention includes a method for amelioration and/or establishment of treatment and/or management of uncontrolled electrical conductance and/or conditions related to such abnormal conductance and/or supposed to be or likely to arise from same conditions and/or seizures and/or epileptic seizures and/or convulsions, by administering and/or using such metallic nanoparticle either alone and/or as bimetals and/or in combination preferably with antiseizure/ anti-epileptic drugs and/or as conjugated thereof.

Hence if such metallic nanoparticles, used in any one or more ways as mentioned above, may be adopted for their usage in the treatment of and/or to manage and/or to regulate the pathophysiological/pathological conditions which are supposed to be due to uncontrolled nature of electrical conductance activity and/or in conditions which are expected to lead due to the activity because of uncontrolled electrical conductance and/or similar to or expected to lead the physiological state and/or pathological state and/or seizures and/or epileptic seizures and/or convulsions etc

Claims

1. A pharmaceutical composition comprising at-least one metallic nanoparticles used either alone or in theirs's combination.

a) These nanoparticles may be (but not limited to); Gold (Au), Silver (Ag), Selenium (Se), Zinc (Zn), Copper (Cu), Iron (Fe), Platinum (Pt), Palladium (Pd), Manganese (Mn), Ruthenium (Ru), Indium (In), Iridium (Ir), Cobalt (Co), Titanium (Ti), Zirconium (Zr), Hafnium (Hf), Vanadium (V), Niobium (Nb), Tantalum (Ta), Chromium (Cr), Molybdenum (Mo), Tungsten (W), Osmium (Os), Nickel (Ni), Rhodium (Rh) etc.

b) These nanoparticles may be either in their nobel form (zero valent forms) or bimetallic form or oxide forms or conjugate form or as complexes, or in the form which involve the process such as ionic bonding or co valent bonding.

c) These metals may also be in any other forms of metals which would change into nano-size during in- vivo dose administration.

d) These nanoparticles of metals may be also present as doped metals inside any inorganic, organic or hybrid type of composite materials.

e) As mentioned in preceding claim, wherein mean size of such metallic nanoparticles should fall below 1000 nm, more precisely 1 nm - 1000 nm , or more preferred to say that mean particle size is below 300 nm. While, Shape of these nanoparticles may be identical to any geometrical forms which may vary from spherical or triangular or cubical or tetrahedral or tetragonal, while their preferred distribution is relatively monodisperse in nature (in a broad sense).

f) As in claim above, dose of any such metallic nanoparticles in the composition ranges preferably above the ^g and up to 1000 mg, or 10 μg-1 mg , or 100 μg - 900 μg or 150 μg - 500 μg. However, such dose of metallic nanoparticles may also be preferably lower than their respective LD-50 dose.

2. These nanoparticles may be synthesized by providing some source of external energy which may suitably involve thermodynamic process or may follow the chemical reaction in presence of at-least one reducing agents which are sourced either from chemical reaction or may be sourced from semisynthetic route or obtained from nature itself. Alternative method of synthesis may also involve process using any living system as a bioreactor or to provide reducing environment, such system at least may be any one of the plant and their culture, microorganisms, animals etc.

a) These nanoparticles, may be either functionalized or capped or stabilized with at least one pharmaceutically acceptable excipients which would help to incorporate at least one of the features in the final formulation such as higher stability, better delivery profile or enhanced efficacy or it may also work as spacer group so that it would carry and deliver any moiety, if present together, to the site of action (for example glucose like molecules for facilitating of attached molecules to the Brain, b) Such pharmaceutical composition above here, may have other pharmaceutically acceptable excipients selected from the group comprising of diluents, lipids, surfactants, penetration enhancer, mucoadhesive, dyes, binders, disintegrating agent, colorants, lubricants, plasticizers, coating agents, opacifiers, antioxidants, flavouring agents, taste masking agents, buffers, stabilizing agents, polymers etc., either alone or in combination thereof.

3. Here, we also claim gum ghatti (Preferred source: Anoaeissus latifolia) as preferred reducing agent due to its inherent reducing properties, its activity as stabilizing agent as well as capping agent, using microwave oven as preferred source of energy.

4. Here, we also claim first medical use of atleast one of the above mentioned metallic nanoparticles or their combination in the manufacture of a medicament for the therapeutic and/or prophylactic treatment of pathological or physiological conditions, including at-least any one of neurosignaling disorders and/or neurotransmitter disorders and/or neurodegenerative conditions which more specifically may be (but not limited to) epilepsy, seizures, convulsions, Parkinsonism etc.

5. We also claim here, composition of metallic nanoparticles, preferably with at-least one of anti-epileptic/anti-seizure drugs which may include (but not limited to); Phenobarbitone, Primidone, Phenytoin, Fosphenytoin, Ethosuximide, Sodium Valproate, Valproic acid, Carbamazepine, Oxcarbazepine, Lamotrigine, Levetiracetam, Vigabatrin, Tiagabine, Lacosamide, Zonisamide, Retigabine, Safinamide, Topiramate, Gabapentin, Pregabalin, Diazepam, Lorazepam, Clobazam, Clonazepam e.t.c, and/or anti-oxidant and/or nutraceuticals and/or other therapeutic active substances which may exert or supposed to exert any protective effects against pathological or physiological condition on their prophylactic or therapeutic usage against any neurosignaling disorders or neurotransmitter disorders or neurodegenerative conditions which may be but not limited to, epilepsy, convulsions, seizures, Parkinsonism etc. The drugs mentioned here may be either in their pharmaceutically acceptable salts, esters, hydrates, derivatives or prodrugs thereof accordingly.

6. We further claim here, A preferred pharmaceutical composition,

a) wherein, metallic nano formulation is of gold and the anticonvulsant/anti-epileptic drug is carbamazepine. b) We further claim here, A preferred pharmaceutical composition wherein, the metallic nanoformulation is of silver and the anticonvulsant/anti-epileptic drug is carbamazepine.

c) We further claim here, A preferred pharmaceutical composition wherein, metallic nanoformulation is of zinc and the anticonvulsant/anti-epileptic drug is carbamazepine.

Here, dose of such metallic nanoparticles is preferably atleast ^g to 1000 mg, or 10 μg- 1 mg, or 100 μg - 1000 μg or 150 μg - 500 μg ; as well as, the amount of anticonvulsant/anti-epileptic drugs is atleast above 1 mg but below their LD-50 value (preferred equivalent dose of carbamazepine 2 - 100 mg/250 g body weight of mice).

7. We also claim here, a synergistic and potentiating formulation of above mentioned composition of metallic nanoparticles along with any drugs (as mentioned in above category) preferably in their therapeutic effective dose range which may be used in the mammals for treatment of epilepsy, seizures, convulsions or other electrical conductance related disorders in need thereof

8. As claimed above, any such composition may be further designed as encapsulated delivery system or matrix delivery system or a hybrid of both, so that release of active constituents be either in controlled manner or follow fast release pathway or may be mixed of both to deliver such preparation by means of either any sophisticated machine or any carriers using novel delivery system approach.

9. Such above mentioned any composition may also formulated as tablets, capsules, powers, syrups etc. as well as fabricated in such a way so that to fulfil the purpose for meeting the criteria of either implantable, topical, injectable or orally or inhalation or nasal type delivery system but not limited to these.