DESC:Technical field:
The present invention relates to synergistic compositions of bioactive agents that improve neuronal function by repairing damaged nerves and simultaneously controlling nerve pain and inflammation.

Particularly, the invention relates to potent bioactive compositions comprising of synergistic combination of fatty acid amide and pyrimidine nucleotides along with pharmaceutically acceptable excipients; wherein the fatty acid amide is palmitoylethanolamide (PEA), the pyrimidine nucleotides are cytidine-5'-monophosphate (CMP) and uridine 5'-monophosphate (UMP).

Further the invention provides synergistic composition for treating or controlling neuropathic pain, moreover the composition is useful for treatment of polyneuropathies, neuritides, myopathies, complex neuropathies, chronic inflammation, demyelinating polyneuropathy diabetic polyneuropathies, multiple sclerosis, spinal syndromes, and neuralgia.

Background and Prior art:
Pain is felt when the nerve endings sense that something is wrong with the body and send pain signals to the spinal cord and brain. Neuropathic pain occurs when the nerves themselves are damaged by disease or injury, causing them to misfire and send pain signals to the brain.

Neuropathic pain is a chronic pain condition. It’s usually the result of, or accompanied by, an injury, disease, surgery or infection. Neuropathic pain is caused by damage or inflammation of the nervous system. It is a complex syndrome. Particularly it is caused by damage to the somatosensory nervous system. The damage can be due to an illness or injury, and may affect the brain, spinal cord, or the nerves in the rest of the body (such as face, arms, legs and torso).

There are several known causes for neuropathic pain, which include injury; spinal cord damage; diabetes; nerve compression (trapped nerve), such as in carpal tunnel syndrome or sciatica; nerve invasion by a tumor; infections such as shingles and HIV/AIDS; multiple sclerosis; stroke; some types of chemotherapy; and vitamin B12 or vitamin B1 (thiamine) deficiency.

Neuropathic pain is a kind of pain that escalates with time. It can be acute or chronic. Chronic manifestations commence with subtle pain and develop gradually. On the contrary, pain in an acute form develops spontaneously, advances quickly, and resolves slowly as the injured nerve cells begin to heal.

Nerves are specialized cells, which carry messages from one part of the body to another, in the form of tiny electrical signals. Nerves are made up of cable-like bundles of nerve cells (called neurons) and each neuron has three main parts, which include dendrites; cell body; and axon.

The dendrites receive impulses from sensory receptors or other neurons and send them towards the cell body, which contains the nucleus.

Impulses are then conducted along the full length of the axons such that the impulses move away from the cell body to connect with the dendrites of another neuron, muscle, organ or gland of some kind.
NeuronsNeurons are the basic building blocks of the nervous system. These specialized cells are the information-processing units of the brain responsible for receiving and transmitting information. Each part of the neuron plays a role in communicating information throughout the body.

Neurons carry messages throughout the body, including sensory information from external stimuli and signals from the brain to different muscle groups in the body. The unique structure of the neuron allows it to receive and transmit signals to other neurons as well as other types of cells.

The myelin surrounding the neurons protects the axon and aids in the speed of transmission. The myelin sheath is broken up at points known as the nodes of Ranvier or myelin sheath gaps. Electrical impulses are able to jump from one node to the next, which plays a role in speeding up the transmission of the signal.

Myelin is a lipid-rich (fatty) substance formed in the central nervous system (CNS) by glial cells called oligodendrocytes, and in the peripheral nervous system (PNS) by Schwann cells. Myelin insulates nerve cell axons and assembles voltage-gated sodium channel clusters at discrete nodes along its length. This helps to increase the speed of electrical impulses in neurons, thereby increasing the speed at which information (encoded as an electrical signal) travels from one nerve cell body to another (as in the CNS) or, for example, from a nerve cell body to a muscle (as in the PNS).

The myelin sheath is a fatty layer that protects nerve cells, extending to cover the nerve throughout its length, except in the nodes of Ranvier. Myelin assists in accelerating the speed at which an impulse travels along a nerve by causing the impulse to jump from one node of Ranvier to the next. Myelin improves the conduction of action potentials, which are needed to send information down the axon to other neurons. It facilitates conduction in nerves while saving space and energy. Myelin helps to prevent the electric current from leaving the axon. It permits a larger body size by maintaining agile communication between distant body parts.

When the myelin sheath is damaged, nerves do not conduct electrical impulses normally. Damage to the myelin sheath, called demyelination, compromises the nerve's ability to conduct impulses that results in chronic nerve pain which includes multiple sclerosis, optic neuritis, neuromyelitis optica, transverse myelitis, acute disseminated encephalomyelitis, adrenoleukodystrophy and adrenomyeloneuropathy, inherited demyelinating diseases such as leukodystrophy, and Charcot-Marie-Tooth disease. Subacute combined degeneration of spinal cord secondary to pernicious anaemia can lead to slight peripheral nerve damage to severe damage to the central nervous system, affecting speech, balance, and cognitive awareness.

A demyelinating disease is any condition that results in damage to the protective covering (myelin sheath) that surrounds nerve fibers in brain, optic nerves and spinal cord. When the myelin sheath is damaged, nerve impulses slow or even stop, causing neurological problems. Breakdown in myelin resulting in desheathing of the axon has been considered as an eminent factor in neuropathic pain. When myelin degrades, conduction of signals along the nerve can be impaired or lost, and the nerve eventually shrinks.

If the sheath is able to repair and regenerate itself, normal nerve function may return. However, if the sheath is severely damaged, the underlying nerve fiber can die. Nerve fibers in the central nervous system (brain and spinal cord) cannot fully regenerate themselves. Thus, these nerve cells are permanently damaged.

In adults, the myelin sheath can be damaged or destroyed by stroke, inflammation, infections, immune disorders, metabolic disorders, nutritional deficiencies (such as lack of vitamin B12), poisons (such as carbon monoxide), drugs (such as the antibiotic ethambutol), excessive use of alcohol, diabetes complications, etc. Other viruses such as human immunodeficiency virus (HIV), the measles virus, and herpes zoster, the virus associated with chickenpox, can also damage the myelin. Also, lack of blood supply causes demyelination.

Hence, there is a need to repair or regenerate the damaged myelin sheath, so that neurological function in the body can be restored. Besides medications, scientists are examining the potential role of various supplements in the myelin repair. These dietary supplements include Alpha lipoic acid, Curcuma longa, or turmeric, Biotin, Vitamin D, Omega-3 fatty acids, however, these nutrients are found less effective and they shift the body's metabolic demand away from protecting the brain and spinal cord, allowing the immune system to wreak havoc.

It is observed by some researchers that nucleotide based drugs like nucleocytidine monophosphate [CMP] seem to be useful for managing peripheral neurological disorders like trigeminal neuralgia, diabetic neuropathy and lumbosciaticalgia, but its central role remains to be elucidated.

One of the most famous brands ‘Nucleo CMP forte’ contain uridine tri¬phosphate, which together with cytidine monophosphate induce biosyn¬thesis of neuronal glycolipid, phospholipid, RNA and DNA ( J Young Pharm, 2016; 8(1): 12-17).

Further it is reported that Nucleo CMP forte is useful in the regeneration of nerve cells by stimulating the synthesis of phospholipids and sphingolipids (the major components of neuronal cell membranes and myelin sheath). Furthermore, Nucleo CMP forte has an essential role in the activation of Schwann cells (Alfaris AA et al. Volume 1(2): 2018, Vet Sci Med).

Further combination of UTP, CMP and hydroxocobalamin shows analgesic effects in peripheral neuropathic pain secondary to spine structural disorders. It supports nerve cell protein synthesis, myelin sheath synthesis, Myelin basic protein synthesis, etc. (M. A. Mibielli et al. / Pain Studies and Treatment 2 (2014) 6-10).

In addition to above literatures there are some patents which disclose therapeutic effect of pyrimidine nucleotides.

EP1032402B1 discloses use of citicoline for the preparation of a medicament for treating multiple sclerosis.

US5583117A relates to the use of acyl derivatives of uridine and cytidine to treat physiological and pathological conditions, including treatment of liver disease or damage, cerebrovascular disorders, bone fractures, respiratory distress syndromes, senile dementias, cardiac damage, and other clinical conditions.

Currently, there are few active substances that affect the physiological regeneration of peripheral nerves. These active ingredients can include pyrimidine nucleotides (uridine monophosphate and cytidine monophosphate). These naturally derived substances play an important role in the synthesis of phospholipids and glycolipids of neuron membranes, affecting the synthesis of myelin membranes and accelerate the regeneration of nerve fibers.

Indeed, dietary pyrimidines (mainly cytosine and uracil derivatives) are able to pass into the circulation, where they are salvaged starting from their nucleosides (cytidine and uridine, respectively).

It is observed that pyrimidine nucleotides show significant effect on regeneration of nerves, however infiltration of immune cells into the site of injury in response to damage to the PNS or CNS may cause inflammation to nervous system. The excessive inflammation in both the peripheral and central nervous system, may contribute to the initiation and maintenance of persistent neuropathic pain.

‘Neuroinflammation’ is detrimental when it manifests itself as multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), various types of dementia, Huntington's disease, or other diseases. A wide range of neurodegenerative diseases, including those affecting the CNS, such as Alzheimer's disease (AD), Parkinson's disease (PD), ALS, and MS, are associated with chronic inflammation. Although inflammation may not be the initiating factor, emerging evidence suggests that sustained inflammatory responses involving microglia and astrocytes contribute to disease progression.

To reduce the extent of neuroinflammation, anti-inflammatory agents such as NSAIDs are usually prescribed but there are several adverse effects associated with administration of synthetic drugs, therefore there is a need for biologically active, naturally derived agents that can considerably reduce the nerve damage induced inflammation.

In the past decades, fatty acid amides were found to be effective to relieve the chronic inflammatory and neuropathic pain. Fatty acid amides include anandamide (N-arachidonoylethanolamine), oleamide, palmitoylethanolamide (PEA), and oleoyl ethanolamide (OEA).

Among fatty acid amides, PEA belonging to the N-acylethanolamines family, exhibit efficacious results in patients with chronic pain associated to a variety of pathological conditions.
Therapeutic activities of PEA along with other agents are cited in some patent applications.

WO2016146453A1 relates to composition comprising palmitoylethanolamide (PEA) and vitamin B for use as an analgesic pharmaceutical in the alleviation of neuropathic pain.

US8663701B2 discloses combination of PEA and antioxidant to control of the peripheral neuropathic pain.

EP2985037B1 discloses a pharmaceutical composition containing palmitoylethanolamide and cytidine-diphosphocholine for the treatment of pathologies of the central nervous system of a traumatic, vascular, degenerative nature, associated with neurodegeneration.

US9512091B2 discloses oxazoline combination with palmitoylethanolamide, for use in the therapy of inflammatory diseases.

There are limited drugs or dietary supplements reported in the art which are effective in neuroregeneration as well as neuroinflammation; much research still needs to be done to optimize the environment for better functioning of nervous system.

Efforts to solve the problem of patient response and improved efficacy of drug therapy has become a primary focus of nervous system treatment. Also, often what is clearly needed is a safe and effective pharmaceutical combination designed for long-term treatment in patients exhibiting nervous system disorders.

By exploring potential areas of neuropathic pain associated with pain control and nerve regeneration, the present inventors have developed efficient nutritional remedy comprising combination of bioactive molecules that synergistically result in improvement in nerve health by ameliorating neuroregeneration and simultaneously controlling neuroinflammation.

Objective:
The primary object of the invention is to provide a synergistic nutritional composition for improving nervous system.

Further object of the invention is to provide potential and therapeutically effective outcome in the treatment of neuropathic pain.

Another object of the invention is to provide synergistic bioactive compositions comprising active nutrients that significantly reduce neuropathic pain without any adverse effects.

Yet another object of the invention is to provide bioactive compositions that give synergistic effect for alleviating nerve pain by repairing damaged nerves and simultaneously controlling the induced neuro inflammation.

Another object of the invention is to provide synergistic nutrients remedy for treating disorders related to nerve damage induced neuropathic pain.

Summary:
To meet the above objectives, the inventors of the instant invention carried out thorough experiments to establish significant effect of the active nutrients present in the composition that improve nerve damage induced neuropathic pain in a subject in need thereof.

In an aspect, the invention relates to synergistic bioactive composition comprising naturally derived moieties for reliving neuropathic pain.

In another aspect, the invention relates to synergistic nutritional composition comprising of synergistic combination of pyrimidine nucleotide and fatty acid amide along with pharmaceutically acceptable excipients; wherein the fatty acid amide is palmitoylethanolamide and the pyrimidine nucleotide is selected from the group consisting of cytidine and/or uridine and phosphate adduct thereof.

In another aspect, the invention relates to bioactive composition which is composed of synergistic combination of palmitoylethanolamide (PEA) and cytidine monophosphate (CMP), optionally uridine monophosphate (UMP), which are present in therapeutically effective amount.

Further the composition provides significant neuropathic pain relieving effect with improved bioavailability and efficacy.

In another aspect, the instant invention provides synergistic nutritional compositions comprising combination of palmitoylethanolamide and cytidine monophosphate and optionally uridine monophosphate for treating a subject suffering from nerve damage induced neuropathic pain.

In yet another aspect, the invention relates to synergistic compositions comprising combination of palmitoylethanolamide present in the range of 10 to 3600 mg, and cytidine monophosphate which is present in the range of 1 to 250 mg, optionally uridine monophosphate which is present in the range of 1 to 250 mg, along with pharmaceutically acceptable excipients/carriers.

In yet another aspect, the present invention provides bioactive compositions that significantly manage nerve damage induced neuropathic pain, wherein the active moieties synergistically regulate neuroregeneration and neuroinflammation.

Further aspect, the invention discloses synergistic composition for treating or controlling neuropathic pain, moreover the composition is useful for treatment of polyneuropathies, neuritides, myopathies, diabetic polyneuropathies, multiple sclerosis, spinal syndromes, and neuralgia.

Abbreviations:
CMP: cytidine monophosphate
UMP: uridine monophosphate
PEA: Palmitoylethanolamide
UTP: uridine triphosphate
PNS: peripheral nervous system
CNS: central nervous system

Brief description of figures:
Fig.1 depicts modulatory effect of test substances on Tail Immersion Latency reaction time at 180 mins- G1-Pain Control; G2-Bio-optimized PEA; G3-CMP; G4-UMP; G5-Bio-optimized PEA+ CMP; G6-Bio-optimized PEA+ UMP; G7-CMP+UMP; G8-Bio-optimized PEA+ CMP+ UMP (PALMINUCLEOTIDE™); G9-Pregabalin (Lyrica® 75 mg).
Fig.2 depicts effect of test substances on percentage maximum possible effect- G1-Pain Control; G2-Bio-optimized PEA; G3-CMP; G4-UMP; G5-Bio-optimized PEA+ CMP; G6-Bio-optimized PEA+ UMP; G7-CMP+UMP; G8-Bio-optimized PEA+ CMP+ UMP (PALMINUCLEOTIDE™); G9-Pregabalin (Lyrica® 75 mg).

Detailed Description:
The invention will now be described in detail in connection with certain preferred and optional embodiments, so that various aspects thereof may be more fully interpreted and comprehended. However, any skilled person or artisan will appreciate the extent to which such embodiments could be generalized in practice.

It is further to be understood that all terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting in any manner or scope.

Unless defined otherwise, all technical and scientific expressions used herein have the same meaning as commonly understood by one of ordinary skill in the art to which embodiments of the invention pertain.

In describing and claiming the embodiments of the present invention, the following terminology will be used in accordance with the definitions set out below which are known in the state of art.

The singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Also the term ‘composition’ does not limit the scope of the invention for multiple compositions that can be illustrated for best mode of the invention.

The term “pharmaceutically/nutraceutically acceptable salt,” as use herein, represents those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response and the like and are commensurate with a reasonable benefit/risk ratio. Particularly the term “pharmaceutically-acceptable salts” refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds, alkali or alkaline earth metal salts, as well as solvates, co-crystals, polymorphs and the like of the salts.

In preferred embodiment, the invention provides synergistic bioactive composition(s) for treating neuropathic pain, wherein the composition comprising exogenous therapeutic blend of fatty acid amide and pyrimidine nucleotides along with pharmaceutically acceptable excipients.

The term ‘bioactive’ is related to any compound present in the foods of humans, animals, or plants that influence physiological or cellular activities in the organism consuming it. The bioactive compounds include flavonoids, carotenoids, carnitine, choline, coenzyme Q, creatine, dithiolthiones, phytosterols, polysaccharides, phytoestrogens, glucosinolates, polyphenols, anthocyanins, prebiotics, taurine, vitamins, enzymes, proteins, amino acids, dietary nucleotides, nucleosides, peptides antibiotic and like thereof.

The term bioactive composition can be interchanged with ‘nutritional synergistic composition’ or ‘composition comprising biologically active compounds or biomolecules’.

In another embodiment, the invention discloses synergistic bioactive composition, wherein the fatty acid amide moiety is palmitoylethanolamide (PEA), for controlling nerve pain and inflammation; wherein the neuroinflammation is regulated by reducing or suppressing response to stimulation of innate immune system cells.

PEA, an endogenous fatty acid amide, is a congener of the endocannabinoid anandamide (N-arachidonoylethanolamine or AEA) that belongs to a class of lipid mediators, the superfamily of N-acylethanolamine (NAE) endogenous biologically active lipids including the endogenous cannabinoid receptor ligand anandamide and the satiety factor oleoylethanolamide.
PEA is a natural compound, and found in a variety of food products, such as soybean, lecithin, egg yolk, and peanut meal. The chemical formula (I) of PEA is as below:

(I)
PEA can also be denoted as ‘Hydroxyethylpalmitamide’, ‘Palmidrol’, ‘N-Palmitoylethanolamine’, and ‘Palmitylethanolamide’.

In another embodiment, the PEA used in the composition is present in the combination of suitable solubilizer or bioenhancer to enhance the solubility and bioavailability of poorly water soluble PEA, the efficacy of micronized PEA is improved under optimum condition by using bio-enhancers, that are collectively termed as ‘Bio-optimized PEA’.

In additional embodiment, the micronized PEA can be incorporated in a matrix to form micelles, inclusion complex, encapsulates.

Moreover, the nerve damage or injury or crush induces inflammation due to activation of innate immune system cells such as mast cells, glial cells, astrocytes and microglia, and this activation can cause neuropathic pain in the peripheral and central nervous system. Particularly activation of two principle innate immune cell populations microglia and mast cells, leads to neuroinflammation.

Glias are non-neuronal cells that maintain homeostasis, form myelin, and provide support and protection to neurons in both central and peripheral nervous systems. They also play an important role in the synthesis, release, and uptake of various neurotransmitters.

In another embodiment, the invention provides PEA as active moiety that control the activation of mast and glia cells interaction which are responsible for neuronal pain sensitization caused due to nerve injury or damage.

The main etiopathogenesis of neuroinflammation is glia–mast cell neuro-immune interaction that contributes to promotion of pain transmission pathways. The therapeutic potential of naturally occurring fatty acid ethanolamides, preferably palmitoylethanolamide controls systemic inflammation or blockade of signalling pathways from the periphery to the brain by modulating the activation of non-neuronal cells and thereby regulating neuronal sensitization-both peripherally and centrally.

In another embodiment, the invention provides significant role of PEA as mast cells stabilizer. Nervous injury triggers neuroinflammation, which activates mast cells, and activated mast cells in turn release inflammatory factors, such as bradykinin, prostaglandins, histamine, and substance P. The neuropathic pain induces mast cell activation, degranulation and neutrophil and macrophage infiltration, which is reversed by treatment with a mast cell stabilizer.

The efficacy of PEA in this group confirms that its actions are independent from those of other therapies and confirms that the concomitant control of mast cell activity in the periphery and microglia activation in the CNS significantly reduces the intensity of neuropathic pain.

In another embodiment, the PEA can be used in various forms including but not limited to non-micronized form (nm-PEA), micronized form (m-PEA), or ultra-micronized form (um-PEA). It is observed that PEA, when administered in an appropriate form e.g., micronized and/or ultra micronized indicates significant pain relief.

In another embodiment, the PEA is present in the micronized form with improved bioavailability and solubility.

In some embodiment the composition comprising micronized palmitoylethanolamide (m-PEA) having particle size in the range of 2 µm -10 µm, preferably 2µm -6µm.

In another embodiment, the synergistic bioactive composition comprises therapeutically effective amount of PEA and pharmaceutically acceptable salts thereof, wherein PEA is present in the range of 1-3600 mg, preferably in the range of 5-1000 mg, more preferably in the range of 5-650 mg, of total composition.

In another preferred embodiment, the invention provides the bioactive composition comprising pyrimidine nucleotides, wherein the pyrimidine nucleotides are selected from the group consisting of cytidine monophosphate, cytidine diphosphate, cytidine triphosphate, uridine monophosphate, uridine diphosphate, uridine triphosphate either alone or combination thereof.

Preferably pyrimidine nucleotide is cytidine monophosphate; which is also known as cytidine 5'-monophosphate, cytidylic acid; 5'-cytidylic acid; cytidine 5'-monophosphate; cytidine 5'-phosphate; cytidylate; 5'-CMP or simply cytidylate.

Further, cytidine monophosphate salt is preferably sodium salt, which can be also referred as cytidine 5'-monophosphate disodium salt; sodium ((2R,3S,4R,5R)-5-(4-amino-2-oxopyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl phosphate; disodium CMP; CMP disodium salt; cytidine-5'-monophosphate disodium salt.

‘Cytidine monophosphate or CMP’ is a nucleotide of cytosine. It is an ester of phosphoric acid with the nucleoside cytidine. CMP consists of the phosphate group, the pentosesugar ribose, and the nucleobase cytosine; hence, a ribonucleoside monophosphate. The chemical formula (II) of Cytidine 5'-monophosphate disodium salt is as below:

(II)

Cytidine is a nucleoside molecule that is formed when cytosine is attached to a ribose ring (also known as a ribofuranose) via a ß-N1-glycosidic bond. Cytidine is a component of RNA. Cytidine 5'-monophosphate (CMP) is used as a substrate of uridine monophosphate (UMP)/cytidine monophosphate (CMP) kinase to form Cytidine Diphosphate (CDP) which upon phosphorylation to Cytidine triphosphate (CTP) supports DNA and RNA biosynthesis.

The food sources of cytidine include beer, breastmilk, barley, baker’s yeast, sugar cane extract, and vegetables. The monophosphate portion of cytidine is the transporter that assists cytidine to move through the gastrointestinal tract unharmed and to cross the blood-brain barrier (BBB).
It is observed that nerve injury damages myelin sheath that causing an interruption in conduction of the impulse down the nerve fiber. Most commonly, this involves compression of the nerve or disruption to the blood supply (ischemia).

Peripheral nerve injury has been shown to be associated with statistically significant increases in the requirement for pyrimidine nucleotides, as damaged nervous tissue is unable to synthesize these compounds because of depletion of the stores of the relevant enzymes. The use of pyrimidine nucleotides (particularly cytidine monophosphate or uridine monophosphate) significantly accelerates the regeneration of nerve pathways after nerve tissue damage and also influences the energy-producing metabolic pathways, myelin sheath synthesis, and levels of various neurotransmitters.

In another embodiment, the pyrimidine nucleotide, preferably cytidine monophosphate accelerates myelination of peripheral nerve fibers and promotes their regeneration. It helps to restore myelin sheath which is impaired due to nerve crush or damage.

Cytidine monophosphate is involved in the synthesis of the complex of lipids forming sphingomyelin, which is the main component of the myelin sheath. Uridine triphosphate acts as a coenzyme in the synthesis of glycolipids, neuronal structures, and myelin sheaths, supplementing the action of cytidine monophosphate.

This demonstrates the significant effect of the CMP, and its ability to restore the functional state of myelin and to stimulate maximally the potential for reinnervation.

Regeneration/protection of the myelin sheath after a nerve lesion is crucial element for normal neuron functioning. Nucleotides CMP and UMP have proven to be efficacious in treating the cause of the myelin sheath lesion in several experimental and clinical models. Notably nucleotides CMP and UMP enhance stimulation of myelin sheath synthesis and hence effective in central and peripheral nervous system regeneration.

In another embodiment, the synergistic composition comprises therapeutically effective amount of cytidine monophosphate and pharmaceutically acceptable salts thereof, wherein the cytidine monophosphate is present in the range of 1-250 mg, preferably in the range of 1-100 mg, more preferably in the range of 1-50 mg, of total composition. Preferably cytidine monophosphate salt is sodium (5 CMP Na2).

In another embodiment, the instant synergistic bioactive composition optionally comprises uridine monophosphate as additional pyrimidine nucleotide to enhance neuroregeneration accompanied by cytidine monophosphate, wherein the uridine monophosphate and its pharmaceutically acceptable salts thereof is present in the range of in the range of 1-250 mg, preferably in the range of 1-100 mg, more preferably in the range of 1-50 mg, of total composition. Preferably uridine monophosphate salt is sodium salt (5 UMP Na2).

The chemical formula (III) of Uridine 5'-monophosphate disodium salt is as below:

(III)

Uridine monophosphate is also known as 5' - uridylic acid. It is an ester of phosphoric acid with the nucleoside uridine. UMP consists of the phosphate group, the pentose sugar ribose and the nucleobase uracil. It is a ribonucleoside monophosphate.

In another preferred embodiment, the invention provides a pain relieving synergistic nutritional composition, wherein the composition comprising combination of Bio-optimized PEA and CMP, optionally UMP along with pharmaceutically acceptable excipients.

In some embodiment, the invention provides synergistic effect of bioactive compounds for controlling nerve damage induced neuropathic pain, wherein one moiety cytidine monophosphate in an effective amount enhances nerve regeneration by stimulating myelin sheath synthesis; simultaneously other moiety palmitoylethanolamide moderates pain sensitization and inflammation by suppressing mast and glia cells activation.

In a preferred embodiment, the present invention provides bioactive composition for treating neuropathic pain, wherein the said composition comprises a synergistic combination of bio-optimized PEA and CMP and UMP along with pharmaceutically acceptable excipient.

In one preferred embodiment, the invention provides synergistic bioactive composition, comprising combination of fatty acid amide and pyrimidine nucleotides, wherein fatty acid amide is bio-optimized PEA and pyrimidine nucleotides are cytidine 5’monophosphate and uridine 5’monophosphate.

In another preferred embodiment the invention discloses synergistic bioactive composition comprising combination of bio-optimized PEA and cytidine 5’monophosphate (CMP) and uridine 5’monophosphate (UMP) which are present in the ratio of 1: 0.001: 0.001 to 1: 0.05 : 0.02 respectively.

In another preferred embodiment, the present invention provides a synergistic bioactive composition, wherein the bio-optimized PEA: CMP: UMP are present in the ratio of 1: 0.008: 0.005.

In another embodiment, the present composition comprises micronized, highly bioavailable and soluble form of PEA, which is present in the range of 70% to 99% by weight of the total composition.

In yet another embodiment, the present synergistic bioactive composition comprises effective amount of CMP which is present in the range of 0.5% to 1.5 % by weight of the total composition.

In yet another embodiment, the present synergistic bioactive composition comprises effective amount of UMP, which is present in the range of 0.1% to 1.5% by weight of the total composition.

In another embodiment, the invention provides a pain relieving synergistic nutritional composition comprising combination of palmitoylethanolamide (PEA) and cytidine-5'-monophosphate (CMP) disodium salt and uridine 5'-monophosphate (UMP) disodium salt present in the ratio of 1: 0.001: 0.001 to 1: 0.01 : 0.01 along with pharmaceutically acceptable excipients.

In the present invention, regeneration of damaged nerves (neuroregeneration) and its normal functioning is carried out synergistically; wherein cytidine monophosphate promotes neuroregeneration by rapid production of myelin sheath in the nervous system; simultaneously palmitoylethanolamide control neuronal sensitization by downregulating activation of mast cell-microglial intracellular pathways thereby controlling nerve pain and inflammation.

The term "neuropathic pain" as used herein has its conventional meaning and has been defined by the International Association for the Study of Pain (IASP, 2011) as 'pain caused by a lesion or disease of the somatosensory nervous system'. The IASP further specifies: 'Neuropathic pain is a clinical description (and not a diagnosis) which requires a demonstrable lesion or a disease that satisfies established neurological diagnostic criteria. The presence of symptoms or signs (e.g., touch-evoked pain) alone does not justify the use of the term neuropathic. Neuropathic pain can be divided according to the IASP in two different pain states:
1. Central neuropathic pain, defined by the IASP as 'pain caused by a lesion or disease of the central somatosensory nervous system'; and
2. Peripheral neuropathic pain, defined by the IASP as 'pain caused by a lesion or disease of the peripheral somatosensory nervous system'.

Particularly nerve damage causes an inflammatory response in which cells associated with the nerve release inflammatory mediators such as eicosanoids; these mediators may contribute to the hyperalgesia which results from nerve injury. A better understanding of the mechanisms involved should lead to improved therapies for neuropathic pain.
In another embodiment, the invention provides synergistic composition for treating nerve damage induced neuropathic pain.

Further neuropathic pain conditions include, but are not limited to, diabetic peripheral neuropathy, herpes zoster, post herpetic neuralgia, trigeminal neuralgia, complex regional pain syndrome, reflex sympathetic dystrophy, migraine headache, phantom limb syndrome, neuropathic pain due to chronic disease (multiple sclerosis, HIV, etc), neuropathic pain due to trauma (causalgia), neuropathic pain due to impingement (i.e. sciatica, carpal tunnel, etc.), neuropathic pain due to drug exposure or toxic chemical exposure, neuropathic pain due to infection or post infection, neuropathic pain due to impaired organ function, neuropathic pain due to vascular disease, neuropathic pain due to metabolic disease, neuropathic pain due to cancer or cancer treatment, neuropathic pain due to autoimmune disease, neuropathic low back pain, neuropathic pain due to fibromyalgia, and neuropathic pain with no known cause (idiopathic), progressive multifocal leucoencephalopathy (PML).

According to the invention , “neuropathic pain” is intended to pain caused by post-stroke pain, spinal cord injury-associated pain, complex regional pain syndrome, complex regional pain syndrome, sensory polyneuropathies, central pain syndrome, headaches, joint pain, backaches, sinus pain, muscle pain, sciatic nerve pain, and pain affecting specific parts of the body, such as shoulders, pelvis, hipbone, and neck, and/or pain that is associated with lower back pain, lumbopelvic pain, vision loss, arthritis, shingles, cancer, a demyelinating neuropathy, muscle weakness, muscle stiffness, muscle spasms, chemotherapy induced neuropathy, post herpetic neuropathy or postoperative pain.

In another embodiment, the instant synergistic bioactive combination is useful for treating peripheral neuropathies such as diabetic neuropathy, chemotherapy-induced peripheral neuropathy, carpal tunnel syndrome, sciatic pain (sciatica), osteoarthritis, low-back pain, failed back surgery syndrome, dental pains, neuropathic pain in stroke and multiple sclerosis, chronic regional pain syndrome, chronic pelvic pain, chronic idiopathic neuropathies, spinal cord injury, fibromyalgia, facial palsy, compression neuralgias, postherpetic neuralgia, and vaginal pains.

In additional embodiment, the instant synergistic composition is useful for treating many types of pain besides neuropathic pain which, incidentally, may also be due to the growing appreciation of the role of neuropathic pain in conditions such as arthritis and other infammatory pain conditions as well as visceral pain syndromes including endometriosis, interstitial cystitis and inflammatory bowel disease, chronic kidney diseases.

As used herein, the term “therapeutically effective amount” is intended to mean the amount of a compound of the present invention effective to reduce or eliminate the neuropathic pain by treatment and/or prophylaxis.

As used herein, the term “pharmaceutically acceptable carriers, diluents or excipients” is intended to mean, without limitation, any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, emulsifier, or encapsulating agent, such as a liposome, cyclodextrins, encapsulating polymeric delivery systems or polyethyleneglycol matrix, which is acceptable for use in the subject, preferably humans. Excipients may also include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, fragrances, glidants (flow enhancers), lubricants, preservatives, sorbents, suspending or dispersing agents, sweeteners, surfactant, anticaking agent, food additives, or waters of hydration.

In some embodiment of the invention, the diluents are selected from starches, hydrolyzed starches, and partially pregelatinized starches, anhydrous lactose, cellulose powder, lactose monohydrate, and sugar alcohols such as sorbitol, xylitol and mannitol, silicified microcrystalline cellulose, ammonium alginate, calcium carbonate, calcium lactate, dibasic calcium phosphate (anhydrous/ dibasic dehydrate/ tribasic), calcium silicate, calcium sulfate, cellulose acetate, corn starch, pregelatinized starch, dextrin, ß-cyclodextrin, dextrates, dextrose, erythritol, ethylcellulose, fructose, fumaric acid, glyceryl palmitostearate, magnesium carbonate, magnesium oxide, maltodextrin, maltose, medium-chain triglycerides, polydextrose, polymethacrylates, sodium alginate, sodium chloride, sterilizable maize, sucrose, sugar spheres, talc, trehalose, xylitol, vehicles like petrolatum, dimethyl sulfoxide and mineral oil or the like.

In some embodiment of the invention, the amount of diluent in the nutritional composition/formulation is present in the range of 1 % to 30% by wt. of the total composition/formulation.

In some embodiment, the binder is selected from disaccharides such as sucrose, lactose, polysaccharides and their derivatives like starches, cellulose or modified cellulose such as microcrystalline cellulose and cellulose ethers such as hydroxypropyl cellulose (HPC); hydroxypropyl methyl cellulose (HPMC); sugar alcohols such as xylitol, sorbitol or mannitol; protein like gelatin; synthetic polymers such as polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), starch, acacia, agar, alginic acid, calcium carbonate, calcium lactate, carbomers, carboxymethylcellulose sodium, carrageenan, cellulose acetate phthalate, chitosan, copovidone, corn starch, pregelatinized starch, cottonseed oil, dextrates, dextrin, dextrose, ethylcellulose, guar gum, hydrogenated vegetable oil, mineral oil, hydroxyethyl cellulose, hydroxymethyl cellulose hydroxyl ethylmethyl cellulose, hydroxypropyl cellulose, inulin, cellulose, methyl cellulose, polyvinylpyrrolidone and polyethylene glycol, lactose, liquid glucose, hypromellose, magnesium aluminum silicate, maltodextrin, maltose, methyl-cellulose, microcrystalline cellulose, pectin, poloxamer, polydextrose, polymethacrylates, povidone, sodium alginate, stearic acid, sucrose, sunflower oil, various animal vegetable oils, and white soft paraffin, paraffin, flavorants, colourants and wax.

In some embodiment of the invention, the amount of binder in the nutritional composition/formulation is present in the range of 0.1 to 30% by wt. of the composition/formulation.

Further according to the invention, the lubricant is selected from magnesium stearate, zinc stearate, calcium stearate, glycerin monostearate, glyceryl behenate, glyceryl palmitostearate, hydrogenated castor oil, hydrogenated vegetable oil, light mineral oil, magnesium lauryl sulfate, medium-chain triglycerides, mineral oil, myristic acid, palmitic acid, poloxamer, polyethylene glycol, sodium benzoate, sodium chloride, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc, potassium benzoate or the like.

In some embodiment of the invention, the amount of lubricant in the nutritional composition/formulation is present in the range of 0.1% by wt. to 5.0% by wt. of the total composition/formulation.

The solubilizing agent is selected from polysorbate 80, sodium lauryl sulfate, anionic emulsifying wax, nonionic emulsifying wax, glyceryl monooleate, phospholipids, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, polyoxylglycerides, sorbitan esters, triethyl citrate, vitamin E, polyethylene glycol succinate, microcrystalline cellulose, carboxymethylcellulose sodium, diethanolamine, ethylene glycol palmitostearate, glycerin monostearate, hypromellose, hypromellose, acetate succinate, lecithin, polyethylene alkyl ethers, aluminum oxide, poly(methylvinyl ether/maleic anhydride), calcium carbonate, crospovidone, cyclodextrins, fructose, hydroxpropyl betadex, oleyl alcohol, povidone, benzalkonium chloride, benzethonium chloride, benzyl alcohol, benzyl benzoate, cetylpyridinium chloride, inulin, meglumine, poloxamer, pyrrolidone, sodium bicarbonate, starch, stearic acid, sulfobutylether beta cyclodextrin, tricaprylin, triolein, docusate sodium, glycine, alcohol, self-emulsifying glyceryl monooleate, cationic benzethonium chloride, cetrimide, xanthan gum, lauric acid, myristyl alcohol, butylparaben, ethylparaben, methylparaben, propylparaben, sorbic acid or the like.
In some embodiment of the invention, the amount of solubilizing agent or surfactant in the nutritional composition/formulation ranges from 0.1% to 10%, preferably 0.1% to 5.0% by wt. of the composition/formulation.

In some embodiment, the glidant is selected from colloidal silicon dioxide, magnesium stearate, fumed silica (colloidal silicon dioxide), starch, talc, calcium phosphate tribasic, cellulose powdered, hydrophobic colloidal silica, magnesium oxide, zinc stearate, magnesium silicate, magnesium trisilicate, silicon dioxide or the like.

In some embodiment of the invention, the amount of glidant present in the nutritional composition/formulation ranges from 0.1% by wt. to 5.0% by wt. of the total composition/ formulation.

In some embodiment, the solvent is selected from water, alcohol, isopropyl alcohol, propylene glycol, mineral oil, benzyl alcohol, benzyl benzoate, flavored glycol, carbon dioxide, castor oil, corn oil (maize), cottonseed oil, dimethyl ether, albumin, dimethylacetamide, ethyl acetate, ethyl lactate, medium-chain triglycerides, methyl lactate, olive oil, peanut oil, polyethylene glycol, polyoxyl, castor oil, propylene carbonate, pyrrolidone, safflower oil, sesame oil, soybean oil, sunflower oil, water-miscible solvents, organic polar or non-polar solvents or mixtures thereof.

In some embodiment of the invention, the amount of solvent in the nutritional composition/formulation is used in a quantity sufficient to 100% by wt. of the composition/formulation.

The additional additives include polymer, a plasticizer, a sweetener, and a powdered flavor, preservative, colorant, surfactant and other excipients. The powdered flavor composition includes a flavourant associated with a solid carrier, coating materials are used, for example synthetic polymers, shellac, corn protein (zein) or other polysaccharides, gelatin, fatty acids, waxes, shellac, plastics, and plant fibers and like thereof. In some embodiment of the invention, the amount of additives in the nutritional composition/formulation are used in the range of 1 to 30 % w/w of unit dose.
Notably, the instant synergistic composition is non-hazardous, non-toxic and safe for human consumption without any adverse/side effects, therefore the instant composition can also be used under preventive therapy in healthy subjects.
The present nutritional composition is used to manage pain conditions in the subject in need thereof, means the administration of the remedy is either to prevent occurrence or for treatment of the pre-existing cause of neuropathic pain such as chronic pain.

In another embodiment, the invention provides a method of treating a subject suffering from neuropathic pain disorders or diseases, the method comprising administering to the subject an effective amount of the present synergistic nutritional composition to alleviate nerve damage and inflammation thereof.

The ‘subject in need thereof’ pertains to subject preferably mammal, more preferably humans having pre-existing or onset symptoms of neuropathic pain, like sciatic pain. The subject may be healthy person which can use the composition under preventive therapy.

In therapeutic applications, compositions can be administered to a patient suffering from pain (e.g., neuropathic pain, complex neuropathy, diabetic peripheral neuropathy) in an amount sufficient to relieve the symptoms of pain like discomfort, soreness, tightness, stiffness, fatigue, sleeplessness, weakened immune system, depression, anxiety, stress, irritability, or disability.

The dosage is likely to depend on such variables as the type and extent of progression of the pain (e.g., as determined by the "Pain Ladder" guideline from the World Health Organization), the severity of the pain (e.g. acute, subacute, or chronic), the age, weight and general condition of the particular patient, the relative biological efficacy of the composition selected, formulation of the excipient, the route of administration, and the judgment of the attending clinician.

An effective dose is a dose that produces a desirable clinical outcome by, for example, improving a sign or symptom of pain or slowing its progression. Accordingly, the effective unit dose can be formulated in the range of 100 to 1000 mg, preferably 150-500 mg and administered one to four times a day, based on the intensity of the pain. The oral administration of an effective dose of the present composition controls neuropathic pain significantly.

In another embodiment, the invention relates to synergistic nutritional composition which can be prepared in a manner well known in the pharmaceutical art, and can be administered by a variety of routes, depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical, parenteral, intravenous, intra-arterial, subcutaneous, intramuscular, intracranial, intraorbital, ophthalmic, intraventricular, intracapsular, intraspinal, intracisternal, intraperitoneal, intranasal, aerosol, suppositories, sub-lingual or oral.

In some embodiment, the instant synergistic nutritional composition can be administered to the subject in need thereof, in the form which is suitable for oral use, such as a tablet, capsule (in the form of delayed release, extended release, sustained release, enteric coated release); hard gelatin capsules, soft gelatin capsules in an oily vehicle, granulate for sublingual use, veg capsule, hard cellulose capsule, effervescent or carbon tablets, aqueous or oily solution, suspension or emulsion, encapsulate, matrix, coat, beadlets, nanoparticles, caplet, granule, particulate, agglomerate, spansule, chewable tablet, lozenge, troche, solution, suspension, rapidly dissolving film, elixir, gel, tablets, pellets, granules, capsules, lozenges, aqueous or oily solutions, suspensions, emulsions, sprays or reconstituted dry powdered form with a liquid medium or syrup; for topical use including transmucosal and transdermal use, such as a cream, ointment, gel, aqueous or oil solution or suspension, salve, parch or plaster; for nasal use, such as a snuff nasal spray or nasal drops; for vaginal or rectal use, such as a suppository; for administration by inhalation, such as a finely divided powder or a liquid aerosol; for sub-lingual or buccal use, such as a tablet, capsule, film, spray. Further the composition can be formulated for parenteral use including intravenous, subcutaneous, intramuscular, intravascular, infusion, intraperitoneal, intracerebral, intracerebroventricular, or intradermal.

In a preferred embodiment, the nutritional composition/formulation is formulated for oral administration. Specifically, the solid nutritional compositions, for example, can be in the form of tablets, capsules, pills, hard capsules filled with liquids or solids, soft capsules, sachets, powders, granules, suspensions, solutions or modified release formulations.

In another preferred embodiment, the oral administration of effective dose of the nutritional composition/formulation attenuates pain sensation and inflammation in the subject of need at an early stage of neuropathic pain. The oral administration of the composition controls chronic pain.

In the context of the present invention, the terms “treatment” and the like refer to alleviate, slow the progression, prophylaxis, attenuation, or cure the pre-existing or occurrence of nerve damage induced neuropathic pain. The instant composition is used for treating nerve damage induced neuropathic pain in a subject in need thereof, means either the administration of the remedy to prevent occurrence or pre-existing cause of neuropathic pain.

In the context of the present invention, the terms “treatment” and the like refer to alleviate, mitigate, prophylaxis, attenuate, manage, regulate, modulate, control, minimize, lessen, decrease, down regulate, up regulate, improve, moderate, prevent, inhibit, stabilize, ameliorate or cure, heal the indications of neuropathic pain.

The ‘subject in need thereof’ pertains to subject preferably mammal, more preferably human with pre-existing symptoms of nerve pain or in a subject to prevent occurrence of neuropathic pain.

The therapeutically effective amount of such bioactive will vary depending upon the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the manner of administration and the like, which can readily be determined by one of ordinary skill in the art.

Thus, a "therapeutically effective" amount is an amount that reduces the risk, potential, possibility or occurrence of a disease or disorder, or provides some alleviation, mitigation, and/or reduction of at least one indicator/biomarker (e.g., blood or serum CRP level), and/or decrease in at least one clinical symptom of a disease or disorder (e.g., neuropathic pain as disclosed herein).

Therapeutic (prescription) supplements are generally administered by the oral or parenteral routes for the treatment of indications including neuropathic pain. The therapeutic administration of materials of the present invention may be in conjunction with other therapies.

The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

While in the foregoing specification this invention has been described in relation to certain embodiments thereof, and many details have been put forth for the purpose of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein can be varied considerably without departing from the basic principles of the invention.

All references cited herein are incorporated by reference in their entireties. The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof and, accordingly, reference should be made to the appended claims, rather than to the foregoing specification, as indicating the scope of the invention.

The invention may be further illustrated by the following examples, which are for illustrative purposes only and should not be construed as limiting the scope of the invention in anyway.
This invention may be embodied in other forms or carried out in other ways without departing from the spirit or essential characteristics thereof. The present disclosure is therefore to be considered as in all respects illustrative and not restrictive, the scope of the invention being indicated by the appended claims, and all changes or alterations which come within the ambit of equivalency are intended to be encompassed therein.

EXAMPLES:

Example: 1
i. Composition 1a: Synergistic Blend
Ingredient w/w %
Bio-Optimized PEA 92±7%
Cytidine 5’ monophosphate-CMP 0.75±0.7%
Uridine 5’monophosphate-UMP 0.45±0.7%

The proprietary blend PALMINUCLEOTIDE™ containing Bio-Optimized PEA™ % + CMP + UMP
The therapeutic proprietary blend with the proportionate excipients filled in soft gel, hard gel, soft cellulose or hard cellulose veg capsule by known technique. Further the blend with the proportionate excipients is compressed to get tablet in coated or uncoated form.

ii. Composition 2: Tablet / Capsule
Ingredient w/w% unit dose
Bio-Optimized PEA 70 -99%
Cytidine 5’monophosphate-CMP 0.5-1.5%
Uridine 5’monophosphate-UMP 0.1-1.5%
Excipient 10-20%
Average Wt 100%
Average wt in mg 300-400

iii. Composition 3: Tablet / Capsule
Ingredient w/w% unit dose
Bio-Optimized PEA 98 ±1%
Cytidine monophosphate-CMP 0.8± 0.1%
Uridine monophosphate-UMP 0.5± 0.1%
Diluent 1-30%
Binder 0.1-30%
Glidant 0.1-5.0%
Lubricants 0.1-5.0%
Additives 1-30%
Solvent QS

iv. Composition 4: Tablet / Capsule
Ingredient mg per unit dose
Bio-Optimized PEA 300
Cytidine 5’ monophosphate-CMP disodium salt 2.5
Uridine 5’ monophosphate-UMP disodium salt 1.5
Microcrystalline cellulose 1-10
Silicon dioxide 1-10
Hydroxypropyl methylcellulose 1-10
Zinc Stearate 1-10
PVP K-30 1-10
Talc 1-5
Magnesium stearate 1-10
Sodium lauryl sulphate 1-10
Manitol 1-15
Propylene Glycol QS
Water QS
Average weight 300-350mg

v. Composition 5: Tablet / Capsule
Ingredient mg per unit dose
Bio-Optimized PEA 300
Cytidine 5’monophosphate-CMP disodium salt 2.5
Uridine 5’ monophosphate-UMP disodium salt 1.5
Microcrystalline cellulose 2-20
Silicon dioxide 1-10
Hydroxypropyl methylcellulose 2-20
Magnesium stearate 2-10
PVP K-30 2-10
Talc 1-20
Polysorbate 80 5-20
Manitol 2-20
IPA QS
Water QS
Average weight 350-400 mg

The present composition is stable for 06 months under the accelerated condition [40°C, 75% RH], where the purity of the active ingredients is above 98% as on dried basis.

Example 2:
Animal Study
Evaluation of analgesic potential of the test substances against tail immersion model in experimental rat.

Animal House conditions-
Lighting: 12 / 12 hour light-dark cycle
Temperature: 22 ± 3 °C
Relative Humidity: 30 to 70%
Animals had continuous access to fresh, potable, uncontaminated drinking water.
Feed: Normal chow diet
Each animal was marked by picric acid and numbering was given individually to each animal. Each cage was numbered separately to identify the group. In each cage single animal was housed in standard stainless-steel cage having facilities for pelleted food and drinking water in bottle.
The study was approved by Institutional Animal Ethical Committee (IAEC) of Radiant Research Services Pvt. Ltd.

Test System and Animal Husbandry
Species: Rat
Strain: Wister
Sex: Male
No. of animals: 54 Animals (n=6 per group)
Body weight: 180-200 gm
CPCSEA Registration Number-1803/PO/RcBi/S/2015/CPCSEA

Group, Designation and Dose Levels

Table 1: Animal grouping and treatment details
Group Treatment Dose No. of Animals
G1 Pain Control Normal saline 6
G2 Bio-Optimized PEA 372 mg/kg 6
G3 CMP 15.5 mg/kg 6
G4
UMP 18.6 mg/kg 6
G5 Bio-Optimized PEA+ CMP 372 mg/kg+15.5 mg/kg 6
G6 Bio-Optimized PEA+ UMP 372 mg/kg+18.6 mg/kg 6
G7 CMP+UMP 15.5 mg/kg +18.6 mg/kg 6
G8 Bio-Optimized PEA + CMP+UMP 372 mg/kg+15.5 mg/kg+18.6 mg/kg 6
G9 Pregabalin (Lyrica®-75mg) 20 mg/kg
6

Experimental Procedure:
Male Wister rat weighing between 180-200 gm were used. Rat were placed into individual cylindrical mice holders leaving the tail hanging out freely. The animals were allowed to get acclimatized to rat holders for 30 min before testing. The lower 5 cm portion of the tail was marked. This part of the tail was immersed in a cup of freshly filled water at 55°C. The reaction time was recorded using a stopwatch. After each determination the tail was carefully dried. The reaction time was determined before and periodically 0 min, 30 min, 60 min, 90 min, 120 min, and 180 min after the oral administration of test substance. The cut off time of immersion was 15 second. The percentage maximum possible effect (%MPE) was calculated using the following formula:
{(Post drug latency-Pre drug latency)/ (15-Pre drug latency)} X100.
The significance of in vivo data was analyzed by One-way ANOVA followed by Dunnet test

Result:
Table 2: Modulatory effect of test substances on Tail Immersion Latency reaction time
TAIL IMMERSION LATENCY REACTION TIME (Seconds)
Group
0 Min 30 Min 60 Min 90 Min 120 Min 180 Min
G1 4.50±0.22 4.33±0.21 4.00±0.26* 4.00±0.26* 4.17±0.17* 3.83±0.17*
G2 4.00±0.26 8.50±0.22* 9.83±0.40* 10.83±0.31* 11.17±0.31* 11.33±0.21*
G3 4.00±0.26 8.00±0.26* 9.17±0.17* 9.83±0.17* 10.50±0.22* 10.83±0.17*
G4 4.17±0.17 8.00±0.26* 9.00±0.26* 10.00±0.26* 10.50±0.22* 11.00±0.26*
G5 4.17±0.17 8.50±0.22* 10.00±0.45* 11.00±0.45* 11.50±0.43* 11.67±0.21*
G6 4.00±0.26 8.50±0.22* 10.17±0.17* 11.17±0.31* 11.50±0.34* 11.83±0.17*
G7 4.17±0.17 8.33±0.21* 9.33±0.33* 10.33±0.21* 10.67±0.21* 11.17±0.31*
G8 4.00±0.26 9.17±0.17* 10.83±0.17* 11.67±0.21* 12.00±0.00* 12.67±0.21*
G9 4.00±0.26 8.83±0.40* 10.83±0.31* 11.50±0.22* 11.83±0.17* 12.33±0.21*

*The values were expressed in Mean±SEM. P < 0.05 was considered as statistically significant.
Table 3: Effect of Test substances on percentage maximum possible effect
PERCENTAGE MAXIMUM POSSIBLE EFFECT
Group
0 Min
(%) 30 Min
(%) 60 Min
(%)
90 Min
(%)
120 Min
(%) 180Min
(%)
G1 -- -- -- -- -- --
G2 -- 40.91 53.03 62.12 65.15 66.67
G3 -- 36.36 46.97 53.03 59.09 62.12
G4 -- 35.38 44.62 53.85 58.46 63.08
G5 -- 40.00 53.85 63.08 67.69 69.23
G6 -- 40.91 56.06 65.15 68.18 71.21
G7 -- 38.46 47.69 56.92 60.00 64.62
G8 -- 47.00 62.09 69.73 72.73 78.82
G9 -- 43.94 62.12 68.18 71.21 75.76

Discussion
In the present study analgesic activity was evaluated by tail immersion model. Table 2 and Figure 1 showed the tail immersion of latency reaction time after the administration of test substances at different time intervals. Treatment groups (G2), (G3), (G4), (G5), (G6), (G7), (G8) and standard (Pregabalin) (G9). The study showed significantly increased latency reaction time in tail immersion when compared with Pain control (G1). Treatment groups (G2), (G3), (G4), (G5), (G6), (G7) (G8) and standard Pregabaline (G9) showed significantly increased percentage maximum possible effect when compared with pain control (G1) at different intervals. Percentage maximum possible effect was more in present composition 1a (G8) (78.82%), when compared with other treated groups at 180Mins. Standard Pregabaline (G9) (75.76%), (G6) (71.21%), (G5) (69.23%), (G2) (66.67%), (G7) (64.62%), (G4) (63.08 %) and (G3) (62.12%) (Table 3 and Figure 2).

Conclusion
Overall results concluded that, test substances treated in combination of test group (G2+G3+G4) i.e. G8 shows more percentage maximum possible effect (78.82%) and exhibits better analgesic activity than the other treated groups i.e. G2 to G7 and standard G9.
,CLAIMS:1. A synergistic bioactive composition(s) for treating neuropathic pain comprising an exogenous therapeutic blend of fatty acid amide and pyrimidine nucleotides along with pharmaceutically acceptable excipients.

2. The synergistic bioactive composition as claimed in claim 1, wherein the fatty acid amide is palmitoylethanolamide (PEA) and the pyrimidine nucleotides are selected from a group consisting of cytidine-5'-monophosphate (CMP) and uridine 5'-monophosphate (UMP) along with pharmaceutically acceptable salts thereof.

3. The synergistic bioactive composition as claimed in claim 1, wherein the salt of cytidine-5'-monophosphate (CMP) and uridine 5'-monophosphate (UMP) is sodium.

4. The synergistic bioactive composition as claimed in claim 1, wherein the palmitoylethanolamide (PEA) and cytidine-5'-monophosphate (CMP) and uridine 5'-monophosphate (UMP) are present in the ratio of 1: 0.001: 0.001 to 1: 0.05: 0.02.

5. The synergistic bioactive composition as claimed in claim 1, wherein the amount of PEA is present in the range of 70-99% by weight of the total composition.

6. The synergistic bioactive composition as claimed in claim 1, wherein the PEA is present in micronized form with particle size in the range of 2µm -6µm, with enhanced bioavailability and solubility.

7. The synergistic bioactive composition as claimed in claim 1, wherein the amount of CMP is present in the range of 0.5% to 1.5 % by weight of the total composition.

8. The synergistic bioactive composition as claimed in claim 1, wherein the amount of UMP is present in the range of 0.1% to 1.5% by weight of the total composition.

9. The synergistic bioactive composition as claimed in claim 1, wherein the pharmaceutically acceptable excipients are selected from a diluent, a binder, a surfactant, a lubricant, a glidant, an additive, a solvent or mixtures thereof.

10. The synergistic bioactive composition as claimed in claim 9, wherein the amount of diluent is present in the range of 1 to 30%; the amount of binder is present in the range of 0.1 to 30%; the amount of lubricant is present in the range of 0.1 to 5%; the amount of glidant is present in the range of 0.1 to 5%; the amount of additive is present in the range of 1 to 30%; the amount of surfactant is present in the range of 0.1 to 5%, by weight of the total composition.

11. The synergistic bioactive composition as claimed in claim 1, wherein oral administration of an effective dose of the composition controls neuropathic pain.

12. The synergistic nutritional composition as claimed in claim 1, wherein the effective unit dose for oral administration is in the range of 150 to 500 mg.

13. A pain relieving synergistic nutritional composition comprising combination of palmitoylethanolamide (PEA) and cytidine-5'-monophosphate (CMP) disodium salt and uridine 5'-monophosphate (UMP) disodium salt present in the ratio of 1: 0.001: 0.001 to 1: 0.01: 0.01 along with pharmaceutically acceptable excipients.

14. The pain relieving synergistic nutritional composition as claimed in claim 13, wherein the
amount of PEA is present in the range of 70 to 99% by weight of the total composition.

15. The pain relieving synergistic nutritional composition as claimed in claim 13, wherein the amount of CMP disodium salt is present in the range of 0.5 to 1.5% by weight of total composition.

16. The pain relieving synergistic nutritional composition as claimed in claim 13, wherein the amount of UMP disodium salt is present in the range of 0.1 to 1.5% by weight of total composition.