DESC:Technical Field:

The present invention relates to novel pain relieving fatty acid amide compositions comprising synergistic combination of fatty acid amide and hydrolytic enzymes inhibitors.

Particularly, the present invention relates to potent pain relieving synergistic compositions, composed of fatty acid amide along with two hydrolytic enzymes inhibitors, preferably hydrolytic enzymes inhibitors are fatty acid amide hydrolase (FAAH) inhibitor and N-acylethanolamine-hydrolyzing acid amidase (NAAA) inhibitor present in an effective amount.

More particularly, the invention relates to pain relieving synergistic compositions, wherein fatty acid amide is ‘Palmitoylethanolamide’ (PEA); FAAH inhibitor is ‘Biochanin A’ and NAAA inhibitor is ‘Diacerein’, along with pharmaceutically acceptable excipients.
Further, the instant novel synergistic composition is useful for improving/treating neuropathic pain.

Background and Prior art:

Pain has a profound impact on the quality of life and can have physical, psychological and social consequences. It can lead to reduced mobility and consequent loss of strength, compromise the immune system and interfere with a person’s ability to eat, concentrate, sleep, or interact with others. A World Health Organization (WHO) study found that people who live with chronic pain are four times more likely to suffer from depression or anxiety. The physical and psychological effects of chronic pain influence the course of the disease. Chronic pain can indirectly influence disease outcomes by reducing a person’s adherence to the treatment.

Considering pain as a public health priority helps to explain its strong linkage with social and economic determinants of health. The social epidemiologic evidence suggests strongly that patterns of chronic non-communicable diseases from the local to the global level are strongly determined by the conditions in which people and communities live, work, and play.

The four largest causes of pain are considered to be cancer, osteo- and rheumatoid arthritis, operations and injuries, and spinal problems, which make the etiology of pain a complex, transdisciplinary affair. Pain has multiple, serious consequences including but not limited to depression, inability to work, disrupted social relationships and suicidal thoughts. It can create negative impact on life, lead to serious disabilities and complications, including sleeping problems, insomnia, anxiety, stress and more.

Further, pain is a response, or a signal sent to the brain to indicate a problem in the body. It tells us that something is not right, and we may need some form of medical treatment. Pain can be mild to severe, occasional, pulsating (throbbing) or constant. It can also be described as an aching, burning, prickling or sharp stabbing.

There are usually two obvious pains found in human body – chronic pain and acute pain, wherein chronic pain is long lasting and usually does not reduce with most treatments and is associated with conditions such as osteoarthritis or fibromyalgia. On the other hand, acute pain occurs suddenly but is temporary and associated with anxiety and emotional stress and resolves as the injury heals.

Chronic pain is further classified into two categories:
First category is nociceptive pain, which is caused by damage to the body tissue and is usually described as a sharp, aching, or throbbing pain. This kind of pain can be due to benign pathology; or by tumors or cancer cells that are growing larger and damaging healthy cells by crowding other parts of the body near the cancer site.

Further nociceptive pain arises from various kinds of troubles in tissues, which are reported to the brain by the nervous system. This is the type of pain everyone is most familiar with, everything from bee stings, and burns, and toe stubs, to repetitive strain injury, nausea, tumours, and inflammatory arthritis. Nociceptive pain typically changes with movement, position, and load.

The second category is neuropathic pain, which occurs when there is an actual nerve damage. Nerves connect the spinal cord to the rest of the body and allow the brain to communicate with the skin, muscles and internal organs. Nutritional imbalance, alcoholism, toxins, infections or auto-immunity can all damage this pathway and cause pain. Other conditions which can predispose patients to develop neuropathic pain include diabetes, small fiber or large fiber neuropathy, vitamin deficiencies, cancer, human immunodeficiency virus (HIV), stroke, multiple sclerosis, shingles, and cancer treatments. The neuropathic pain, characteristic of neuropathies caused by traumatic, compressive, dysmetabolic, and infective injuries, is characterized by spontaneous pain, allodynia, and hyperalgesia.

When the sensory system is impacted by injury or disease, the nerves within that system cannot work to transmit sensation to the brain. This often leads to a sense of numbness, or a lack of sensation. However, in some cases when this system is injured, individuals experience pain in the affected region. Neuropathic pain does not start abruptly or resolve quickly; it is a chronic condition which leads to persistent pain symptoms. Although neuropathic pain is thought to be associated with peripheral nerve problems, such as neuropathy caused by diabetes or spinal stenosis, injuries to the brain or spinal cord can also lead to chronic neuropathic pain.

A cannabinoid is one of a class of diverse chemical compounds that acts on cannabinoid receptors in cells that alter neurotransmitter release in the brain. Ligands for these receptor proteins include the endocannabinoids (produced naturally in the body by animals), the phytocannabinoids (found in cannabis and some other plants), and synthetic cannabinoids (manufactured artificially).

Endogenous cannabinoids fall into two classes, esters and amides. The first putative endocannabinoid is anandamide, also known as N-arachidonoylethanolamine (AEA,) which is a partial agonist at both cannabinoid CB1 and CB2 receptors, and the second endocannabinoid is 2-arachidonoylglycerol (2AG).
Both cannabinoid receptors and endocannabinoids, such as 2AG and anandamide, have been shown to control numerous physiological and pathological processes, including the central nervous system (CNS). Thus, regulating endocannabinoid levels in-vivo represents an interesting therapeutic perspective in several CNS-related diseases. Till date four enzymes - Fatty Acid Amide Hydrolase (FAAH), N-Acylethanolamine-hydrolyzing Acid Amidase (NAAA), Monoacylglycerol Lipase (MAGL), a/ß-Hydrolase Domain 6 (ABHD6) - are shown to control endocannabinoid levels in tissues or in intact cells.

In the past few years, fatty acid amides are found to be effective to relieve the chronic inflammatory and neuropathic pain. Fatty acid amides include anandamide (N-arachidonoylethanolamine), oleamide, palmitoyl ethanolamide (PEA), and oleoyl ethanolamide (OEA). N-Acylethanolamines (NAEs) involve a family of lipid molecules existent in animals and plants, one of which is N-palmitoylethanolamide (PEA) that arouses great attention owing to its anti-inflammatory, analgesic, and neuroprotective activities.

Probably due to the fact that PEA is an endogenous modulator as well as a compound in food, such as eggs and milk, no serious side effects or any drug-drug interactions have been reported. Further, the application and potential efficacy and safety of PEA, in the treatment of various syndromes associated with chronic pain which are poorly responsive to standard therapies, are the key features of PEA that makes it superior than other NAEs.

Notably PEA, an endogenous fatty acid amide belonging to the N-acylethanolamines family, exhibits efficacious results in patients with chronic pain associated with a variety of pathological conditions. The signaling lipid PEA is known to activate intracellular, nuclear and membrane-associated receptors and regulate many physiological functions related to the inflammatory cascade. Therefore, it is of high interest in the treatment of neuropathic pain.

It is observed that neuropathic pain tends to exhibit a relatively poor response to traditional analgesics. PEA is a peroxisome proliferator-activated receptor alpha (PPAR-a) ligand that exerts anti-inflammatory, analgesic and neuroprotective actions. PEA is synthesized from phospholipids through the sequential actions of N-acyltransferase and N-acylphosphatidylethanolamine-preferring phospholipase D (NAPE-PLD), and its actions are terminated by its hydrolysis by two enzymes, FAAH and NAAA.

N-palmitoyl-phosphatidyl-ethanolamine (N-APE) is converted into palmitoylethanolamide and phosphatidic acid by a plasma membrane-associated N-acylated phosphatidylethanolamine-phospholipase D (PLD). PEA is broken down to palmitic acid and ethanolamine by FAAH (which also catabolizes other fatty acid amides) as well as the more selective NAAA.

Recent studies with NAAA and FAAH inhibitors put forth that these enzymes are capable of increasing PEA levels in-vivo in inflammatory processes, and were identified as interesting targets for drug discovery research. Thus, PEA hydrolytic enzyme inhibitors could constitute potential therapeutic alternatives in chronic inflammatory and neurodegenerative diseases. Further, their effective consumption does not show any detrimental effects on the body.

Mireille Alhouayek et al. [FASEB J. 29, 000–000, 2015] discloses the effect of NAAA inhibitor for controlling levels of PEA, wherein NAAA inhibitor shows efficient result for controlling PEA levels in the colon during inflammation. There is no synergistic action of FAAH and NAAA blockers evaluated by Mireille.

Further WO2011027373A1 relates to a pharmaceutical composition containing combination of palmitoylethanolamide in the ultra-micronized form with an antioxidant compound, such as quercetin, resveratrol, polydatin, luteolin, tocopherol, and thioctic acid for treating neuroimmunogenic inflammatory diseases.

EP2921167B1 discloses pharmaceutical composition comprising palmitoylethanolamide (PEA) and L-acetylcarnitine (LAC) for controlling the inflammatory and/or neuropathic pain.
US9512091B2 discloses combination of palmitoylethanolamide with oxazoline derivatives for use as an inhibitory modulator of the activity of the FAAH and NAAA enzymes.
S9321743B2
US9321743B2 relates to oxooxetan compounds for inhibiting N-acyl-ethanolamine-hydrolyzing acid amidase (NAAA), wherein this compound is used as therapeutic agent in the treatment of inflammatory diseases.

Further EP2797584 B1 discloses a composition for use in the treatment of pain, inflammation, fever, and osteoarthritis, comprising diacerein and non-steroidal anti-inflammatory agents consisting of flurbiprofen, loxoprofen, zaltoprofen, ibuprofen, naproxen, ketoprofen, dexketoprofen, fenoprofen, benoxaprofen, suprofen, ibuproxam, alminoprofen, dexibuprofen, and indoprofen.

Furthermore, US20140377343A1 relates to pharmaceutical formulations of flurbiprofen and diacerein having anti-inflammatory, analgesic, antipyretic, and osteoarthritis-treating activities.

WO2011123719A2 discloses method of treating or preventing abdominal, visceral or pelvic pain in a patient in need thereof, comprising administering a therapeutically effective amount of oxoacetamide derivaties as FAAH inhibitor.

Additionally, US20070155747A1 claims esters of alkylcarbamic acids as FAAH inhibitors and the processes for the preparation of thereof, whereas WO2017201103A1 discloses piperazinyl methanone analogues as NAAA inhibitors for treatment of a pathological state, including pain, an inflammatory condition, or a neurodegenerative disorder.

It may be noted that most of the compositions cited in the prior arts are administered in presence of nonsteroidal anti-inflammatory drugs (NSAID) drugs. However, NSAIDs are generally not recommended for people with kidney disease, heart failure, or cirrhosis, or for people who take diuretics. Some patients who are allergic to aspirin may be able to take selective NSAIDs safely. Therefore, the above compositions are subject specific and show limited effect. Further nowhere in the art discloses the synergistic composition of NAAA and FAAH inhibitors which are naturally derived to inactivate fatty acid amide hydrolysis.

Therefore, to avoid possible side effects during the administration of pain reliving medicine, there is a requirement to develop potent dietary and bioactive phytochemicals to inhibit the activity of respective enzymes for PEA hydrolysis with no side effect.

The present inventors have successfully developed novel compositions, which are accomplished by rigorous research and experiments. The composition comprises combination of PEA and bioactive phytochemicals that inactivate the activity of hydrolytic enzymes responsible for PEA degradation. Consequently, this hydrolytic enzyme inhibition regulates the level of PEA in the body.

Objective:

The primary object of the invention is to provide synergistic nutritional compositions for relieving chronic pain.

Another object of the invention is to provide synergistic composition comprising bioactive phytochemicals that significantly reducing neuropathic pain without any adverse effects.

Another object of the invention is to provide synergistic PEA composition comprising effective amount of hydrolytic enzyme inhibitors that obviate degradation of PEA.

Yet another object of the invention is to cater nutritional compositions that give synergistic effect for relieving nerve pain by enhancing the cellular concentration of PEA.

Summary:

To meet the above objectives, the inventors of the instant invention have carried out thorough experiments to establish significant effects of the active nutrients present in the composition for improving pain and/or pain related disorders, in a subject in need thereof.

In an aspect, the invention relates to synergistic composition of bioactive phytochemicals for relieving chronic pain.

In another aspect, the invention relates to synergistic fatty acid amide composition comprising of fatty acid amide in combination with fatty acid amide hydrolase (FAAH) inhibitor and N-acylethanolamine acid amidase (NAAA) inhibitor in an effective amount.

In another aspect, the instant invention provides potent pain relieving synergistic compositions, wherein the fatty acid amide is PEA; FAAH inhibitor is ‘Biochanin-A’ and NAAA inhibitor is ‘Diacerein’, along with pharmaceutically acceptable excipients.

In yet another aspect, the invention relates to synergistic compositions comprising combination of PEA, which is present in the range of 1 to 500 mg; Biochanin-A present in the range of 1 to 250 mg, and Diacerein present in the range of 1 to 500 mg along with pharmaceutically acceptable excipients / carriers.

In yet another aspect, the invention relates to synergistic phytochemicals composition, which is particularly useful for treating neuropathic pain.

Abbreviations:
PEA: Palmitoylethanolamide
FAAH: Fatty Acid Amide Hydrolase
NAAA: N-acylethanolamine-hydrolyzing acid amidase
NAE: N-acylethanolamine
AEA: N-arachidonoylethanolamine
ECL: Endogenous Cannabinoid Lipid
GPR55: G protein-coupled receptor 55
TRPV1: Transient Receptor Potential cation channel subfamily V member 1
PPAR-a: Peroxisome proliferator-activated receptor alpha
COX: Cyclooxygenase
iNOS: Inducible nitric oxide synthase
eNOS: Endothelial nitric oxide synthase
2AG: 2-arachidonoylglycerol
CNS: Central nervous system
MAGL: Monoacylglycerol Lipase
ABHD6: a/ß-Hydrolase Domain 6
NAPE-PLD: N-acylphosphatidylethanolamine-preferring phospholipase D
N-APE: N-palmitoyl-phosphatidyl-ethanolamine
PLD: N-acylated phosphatidylethanolamine-phospholipase D
LAC: L-acetylcarnitine
NSAID: nonsteroidal anti-inflammatory drugs

Brief description of figures:

Fig.1 depicts modulatory effect of test substances on Tail Immersion Latency reaction time at 180 mins.

Fig.2 depicts effect of test substances on percentage maximum possible effect G1-Pain Control; G2-Bio-optimized PEA; G3-Biochanin-A; G4-Diacerein; G5-Bio-optimized PEA+ Biochanin-A; G6-Bio-optimized PEA+ Diacerein; G7-Biochanin-A+ Diacerein; G8-Bio-optimized PEA+ Biochanin-A+ Diacerein (RITANURON™); 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.

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, as well as solvates, co-crystals, polymorphs and the like of the salts.

In preferred embodiment, the invention discloses synergistic bioactive phytochemicals compositions, comprising specific combination of therapeutically active phytochemicals, particularly the phytochemical composition is composed of fatty acid amide and NAAH inhibitor and FAAH inhibitor and pharmaceutically acceptable salts thereof, for treating neuropathic pain in a subject in need thereof, wherein the fatty acid amide is Palmitoylethanolamide (PEA); FAAH inhibitor is ‘Biochanin-A’ and NAAA inhibitor is ‘Diacerein’, along with pharmaceutically acceptable excipients.

In one preferred embodiment, the invention provides a synergistic nutritional composition(s) for treating neuropathic pain, wherein the composition comprising therapeutic blend of PEA and Biochanin-A and Diacerein along with pharmaceutically acceptable excipients.

Further PEA and Biochanin-A and Diacerein are present in a specific ratio to give synergistic therapeutic efficacy.

In another preferred embodiment, the invention provides a pain relieving synergistic nutritional composition, wherein the composition comprising combination of PEA and Biochanin-A and Diacerein along with pharmaceutically acceptable excipients.

The present invention is directed to compounds and compositions of NAAA and FAAH inhibitors, which are suitable for treatment of various diseases associated with reduced PEA levels in a cell, organ, or other body structure (or even the entire body). Most preferably, such modulation will result in treatment and/or prevention of neuropathic pain, inflammation, and other disorders in which abnormal N-acylethanolamine levels are associated with a disorder.

N-acylethanolamines (NAEs) are a class of naturally occurring lipid mediators composed of a fatty acid and ethanolamine the so-called fatty acid ethanolamines (FAEs). The principal FAEs include the endocannabinoid N-arachidonoylethanolamine (anandamide), and its congeners N-stearoylethanolamine, N-oleoylethanolamine and N-palmitoylethanolamine (PEA or palmitoylethanolamide).

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). PEA is a natural compound, and found in a variety of food products, such as soybean, lecithin, egg yolk, and peanut meal.

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

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

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

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 bioenhancer 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.

It may be noted that activation of inflammatory cascades and non-neuronal cells, such as glial cells, astrocytes, and mast cells, contributes to the pathogenesis of chronic pain and to peripheral and central sensitization. PEA reduces mast cell migration and degranulation and reduces over-activation of astrocytes and glial cells. Both mast cells and glial cells shift under the influence of PEA from activated immune cells to resting phenotypes. Further on the molecular level PEA reduces the activity of the pro-inflammatory enzyme COX, as well as eNOS and iNOS.

PEA has a number of other properties, related to its affinity for various receptors: the orphan cannabinoid receptors (G protein-coupled receptors like (GPR55) and (GPR119), the vanilloid receptor-transient receptor potential vanilloid 1 (TRPV1) and the nuclear (PPAR-a).

Particularly PEA inhibits mast cell, glial cells and astrocyte activation, as well as Nerve growth factor (NGF)-related inflammation cascades to get relief from nerve sensation.

Further be reported that immune responses are mainly dependent on immune cells (like mast cells, microglia, neutrophils, macrophages, Schwann cells, and T cells) and some of their inflammatory mediators.

Mast cells can modify sensory transmission via a wide spectrum of mediators, including biogenic amines such as histamine and serotonin, cytokines (interleukin-1b (IL-1b) and tumor necrosis factor-a (TNF-a) in particular), enzymes, lipid metabolites, adenosine triphosphate (ATP), neuropeptides, nerve growth factor (NGF), and heparin-most of which can interact with sensory nerve terminals. Sensory neurons, in turn, by releasing neuropeptides may provoke mast cell activation / degranulation.

Mast cell-nerve terminal activity results in nociceptor sensitization, reduced pain threshold at the site of inflammation and, ultimately, dysfunctional pain signaling and hyperalgesia and when it endures, increased responsiveness of nociceptors can also sensitize spinal cord neurons, leading to central sensitization.

It may be noted that, in a normal state, mast cells have granules that contain a variety of bioactive chemicals. 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 and degranulation and neutrophil and macrophage infiltration, which is reversed by treatment with a mast cell stabilizer. Therefore, mast cells are powerful neuropathic pain mediators.

Glial cells mediate pain processing at the spinal level. Sensitization of central somatosensory neurons is responsible for the development of chronic neuropathic pain. Microglia, macrophages found in the brain, interact with neurons at the site of injury or disease and can be activated through exposure to a number of molecules, including pro-inflammatory signals released from mast cells. This interaction between mast cells and microglia regulates peripheral pain signaling. Microglia activation causes pain states by releasing proinflammatory cytokines, chemokines, and proteases.

Astrocytes, the most abundant glial cell type in the CNS, also play a major role in pain facilitation and are fundamental contributors to the neuropathic pain involved in neuroinflammation. When activated, astrocytes release of Interleukin 1beta (IL-1b), Interleukin-6 (IL-6), and Tumor Necrosis Factor-alpha (TNF-a), and prostaglandin E2. Chronic astrocytic activation in nerve injury results in down-regulation of glutamate transporters, ultimately resulting in decreased glutamate uptake and increased excitatory transmission and facilitation of chronic pain.

In another embodiment the instant pain reliving composition of PEA, down-modulates mast cell activation and controls microglial cell activity. Moreover, it acts on the ‘‘roots of the problem’’, i.e., on the cells involved in the generation and maintenance of pain. The administration of instant composition in therapeutically effective amount enhances the PEA level by inhibiting the degradation mediated by the enzymes such as NAAA and FAAH.

Accordingly, the instant PEA composition is used as promising therapeutic target for managing neuropathic pain that mediate mast and glial cell reactivity.

The instant synergistic composition not only down-modulate mast cell activation but also control glial cell behaviors in more effective manner as PEA alone.

The efficacy of PEA composition 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 reduce the intensity of neuropathic pain.
In another embodiment, the synergistic composition comprises therapeutically effective amount of fatty acid amide i.e., PEA or pharmaceutically/nutraceutically acceptable salts thereof, wherein PEA is present in the range of 1-500 mg, preferably in the range of 5-450 mg of total composition or unit dose.

The instant compositions regulate in-vivo level of PEA by inhibiting enzyme degradation in presence of two active enzyme inhibitors ‘Biochanin A’ and ‘Diacerein’. The presence of inhibitors controls the activation of non-neuronal cells and relieving the neuropathic pain in significant manner.

In another embodiment, the instant invention provides FAAH inhibitor or blocker that synergistically controls the deactivation of PEA and maintains its sufficient therapeutic levels in the body that counter the nerve pain, wherein the FAAH inhibitor is ‘Biochanin A’.

‘Biochanin-A’, an O-methylated isoflavone, chemically known as [5,7-Dihydroxy-3-(4-methoxyphenyl) chromen-4-one] is a natural organic compound in the class of flavonoids. Biochanin-A is particularly found in red clover, soy, alfalfa sprouts, peanuts, chick pea (Cicer arietinum) and other legumes. It is observed that ‘Biochanin-A’ exhibits remarkable effect for alleviating neuropathic pain by inhibiting FAAH, the destructive enzyme of PEA.

In another embodiment, the synergistic composition comprises therapeutically effective amount of FAAH inhibitor or pharmaceutically/nutraceutically acceptable salts thereof, wherein FAAH inhibitor i.e., Biochanin A, is present in the range of 1-250 mg, preferably in the range of 1-200 mg of the total composition or unit dose.

In another embodiment the instant invention provides NAAA inhibitor or blocker that synergistically moderates the deactivation of PEA and maintains its sufficient therapeutic levels in the body that counter the nerve pain, wherein the NAAA inhibitor is ‘Diacerein’.

Diacerein (INN), is also known as diacetylrhein, [IUPAC name 4,5-diacetyloxy-9,10-dioxoanthracene-2-carboxylic acid] is insoluble in water. It is a slow-acting medicine of the class anthraquinone.

In the instant composition, Diacerein is potent and selective NAAA inhibitor, which inhibits NAAA enzyme both in the cell-free preparations and intact cells. It acts as a modulator of endogenous PEA that effectively imparts pain relief in a subject in need thereof. It is obtained from a natural source.

In another embodiment, the instant synergistic composition comprises therapeutically effective amount of NAAA blocker or inhibitor i.e., Diacerein and pharmaceutically/nutraceutically acceptable salts thereof, wherein Diacerein is present in the range of 1-500 mg, preferably in the range of 1-400 mg of total composition or unit dose.

In one preferred embodiment, the invention provides synergistic nutritional composition, comprising combination of fatty acid amide and FAAH inhibitor and NAAA inhibitor, which are present in the ratio of 1: 0.001: 0.01 to 1: 0.02: 0.1 respectively.

In yet another preferred embodiment, the invention provides synergistic nutritional composition, comprising bio-optimized PEA and Biochanin-A and Diacerein which are present in the ratio of 1: 0.001: 0.01 to 1: 0.02 : 0.1 respectively, preferably 1: 0.005 :0.02 to 1: 0.01: 0.08.

In another preferred embodiment, the present invention provides a synergistic nutritional composition wherein the ratio of bio-optimized PEA: Biochanin-A: Diacerein is in a range of 1: 0.008: 0.04.

In another embodiment, the present composition comprises micronized, highly bioavailable and soluble form of PEA, which is present in the range of 50-98% by weight of the total composition.

In yet another embodiment, the present composition comprises effective amount of Biochanin-A which is present in the range of 0.1-2.0% by weight of the total composition.

In yet another embodiment, the present composition comprises comprising effective amount of Diacerein, which is present in the range of 1.0-8.0% by weight of the total composition.

In a preferred embodiment, the present invention provides a nutritional composition for treatment of neuropathic pain, wherein the said composition comprises a synergistic combination of bio-optimized PEA and natural FAAH inhibitor and natural NAAA inhibitor along with pharmaceutically acceptable excipient.

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. Some disease entities, such as trigeminal neuralgia, are currently defined by their clinical presentation rather than by diagnostic testing. Other diagnoses such as post-herpetic neuralgia is normally based upon the history.

‘Neurogenic pain’ is also defined as pain due to dysfunction of the peripheral or central nervous system, in the absence of nociceptor (nerve terminal) stimulation by trauma or disease.

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'.
The term "lesion" as used herein has its conventional meaning and refers to a situation when diagnostic investigations (e.g., imaging, neurophysiology, biopsies, lab tests) reveal an abnormality or when there was obvious trauma.

Further neuropathic pain can take a variety of forms depending on its origin and can be characterized as acute, subacute, or chronic depending on the duration.

In certain embodiments, the neuropathic pain include diabetic peripheral neuropathy, post-herpetic neuralgia, postoperative pain, trigeminal neuralgia, phantom limb pain, carpal tunnel syndrome, sciatica, pudendal neuralgia, complex regional pain syndrome, sensory polyneuropathies, mono-neuropathies, or central pain syndrome, headaches, joint pain, backaches, sinus pain, muscle pain, nerve pain, and pain affecting specific parts of the body, such as shoulders, pelvis, and neck, and/or pain that is associated with lower back pain, lumbopelvic pain, arthritis, headache, multiple sclerosis, fibromyalgia, shingles, nerve damage, cancer, a demyelinating neuropathy, chemotherapy induced neuropathy, HIV neuropathy, causalgia, peripheral or polyneuropathic pain, toxic neuropathy, chronic neuropathy caused by chemotherapeutic and antiviral agents, or pruritus induced by uremia, malignancies of various origin, polycythemia, jaundice or cholestasis, iron deficiency, athlete's foot, xerosis, wound healing, thyroid illness, hyperparathyroidism, or menopause.

In another embodiment, the instant synergistic combination is useful for treating peripheral neuropathies such as diabetic neuropathy, chemotherapy-induced peripheral neuropathy, carpal tunnel syndrome, sciatic pain, osteoarthritis, low-back pain, failed back surgery syndrome, dental pains, neuropathic pain in stroke and multiple sclerosis, chronic regional pain syndrome, chronic pelvic pain, 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.
The term ‘phytochemical bioactive’ refers to non-nutrient compounds derived from plants that have biological activity in humans and are considered to be bioactive phytochemicals. These include phenolic compounds, terpenoid compounds, and alkaloid, carbohydrate, lipids, flavonoids, carotenoids and like thereof. They are simple bioactive non-nutrient compounds derived from fruits, vegetables, grains, and other plant foods that are linked to reductions in the risk of major noncommunicable chronic diseases (Journal of Food Science Vol. 78, S1, 2013).

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 chronic pain. The instant composition is used for treating neuropathic pain or pain related disorder in a subject in need thereof, means either the administration of the remedy will prevent the onset or occurrence of pain, or treat pre-existing cause of pain.

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

An “effective amount of bioactive phytochemicals” is therapeutically effective dose of NAAH inhibitor, FAAH inhibitor and PEA to prevent, treat, reduce, and/or ameliorate the symptoms and/or underlying causes of chronic pain or pain related disorders.

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., pain such as neuropathic pain as disclosed herein).
A “pharmaceutically acceptable excipient,” as used herein, refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being non-toxic and non-inflammatory in a patient. Excipients may include, for example, antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, sorbents, suspending or dispersing agents, sweeteners, surfactant, anticaking agent, food additives, solvents or waters of hydration.

In another embodiment, the invention relates to synergistic 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, ophthalmic, aerosol, by suppositories, or oral administration.

Particularly, the composition can be administered to subject in a form 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, effervescent tablets, aqueous or oily solution, suspension or emulsion; for topical including transmucosal and transdermal use, such as a cream, ointment, gel, aqueous or oil solution or suspension, balm, salve, patch or plaster; for nasal use, such as a snuff nasal spray or nasal drops; for vaginal or rectal use, such as a suppositories; for administration by inhalation, such as a finely divided powder or a liquid aerosol; for sub-lingual or buccal use, such as a tablet or capsule, film or for parenteral use including intravenous (IV), intramuscular (IM), subcutaneous (SC) and intradermal (ID), infusion such as a sterile aqueous or oil solution or suspension.

Moreover the term “specific or effective amount” is intended to mean the therapeutically effective dose of instant bioactive compounds namely bio-optimized PEA and Biochanin-A and Diacerein in combination to give significant therapeutic efficacy in the treatment of neuropathic pain, which is otherwise not obtained by use of single ingredient of the composition.
The term “pharmaceutically acceptable salt” refers to a salt prepared from pharmaceutically acceptable non-toxic acids or bases, halides, sulphates, phosphates, nitrate, metal ions, minerals, chelates, complex, esters, oxide, amines which are well known in the art.

As used herein, the term “pharmaceutically acceptable carriers/vehicles/diluents or excipients” is intended to mean, without limitation, any adjuvants, carriers, excipients, sweetening agents, diluents, preservative, dye/colorants, flavor enhancers, surfactants, wetting agents, dispersing agents, suspending agents, complexing agents, stabilizers, isotonic agent, solvent, emulsifier, encapsulating agent, polymers, coating agent, wax, encapsulating polymeric delivery systems. Excipients may also include, antiadherents, antioxidants, binders, pH-modifier, solvents, coatings, compression aids, disintegrants, emollients, , fillers (diluents), film formers, fragrances, glidants (flow enhancers), lubricants, preservatives, sorbents, 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 40% by wt. of the total composition/formulation.

In further 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 0.2 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. The amount of solubilizing agent or surfactant in the nutritional composition/formulation of the present invention 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. The additives 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 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 pain sensation 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 chronic 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, 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 300-600 mg and administered daily once or twice or thrice based on the intensity of the pain.

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, by suppositories, or oral administration.

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, effervescent 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.

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 50-98%
Biochanin-A 0.1-2.0%
Diacerein 1.0-8.0%

ii. Composition 1b: Synergistic Blend
Ingredient w/w %
Bio-Optimized PEA 95±5%
Biochanin-A 0.80±5%
Diacerein 4.0±5%

The proprietary blend RITANURON™ containing Bio-Optimized™ PEA 50-98% + Biochanin-A 0.1-2.0% + Daicerein 1.0-8.0%.

The therapeutic proprietary blend with the proportionate excipients filled in soft gel, hard gel, veg capsule by known technique. Further the blend with the proportionate excipients is compressed to get tablet in coated or uncoated form.

iii. Composition 2: Tablet / Capsule
Ingredient w/w% unit dose
Bio-Optimized PEA 80±5%
Biochanin-A 0.65±5%
Diacerein 3.35±5%
Excipient 10-20%
Average Wt 100%
Average wt in mg 320-400 mg

iv. Composition 3: Tablet / Capsule
Ingredient w/w% unit dose
Bio-Optimized PEA 80±5%
Biochanin-A 0.65±5%
Diacerein 3.35±5%
Diluent 1-20%
Binder 0.2 - 20 %
Glidant 0.1-5%
Lubricants 0.1-5%
Additives 1-10%
Solvent QS

v. Composition 4: Tablet / Capsule
Ingredient mg per unit dose
Bio-Optimized PEA 300
Biochanin-A 2.5
Diacerein 12.5
Microcrystalline cellulose 2-20
Silicon dioxide 1-15
Hydroxypropyl methylcellulose 2-10
Zinc Stearate 2-10
PVP K-30 2-10
Talc 1-10
Magnesium stearate 1-10
Sodium lauryl sulphate 1-20
Manitol 1-20
Propylene Glycol QS
Water QS
Average weight 330-350 mg

vi. Composition 5: Tablet / Capsule
Ingredient mg per unit dose
Bio-Optimized PEA 300
Biochanin-A 2.5
Diacerein 12.5
Microcrystalline cellulose 1-15
Silicon dioxide 1-15
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

vii. Composition 6: Tablet / Capsule
Ingredient mg per unit dose
Bio-Optimized PEA 300
Biochanin-A 2.5
Diacerein 12.5
Microcrystalline cellulose 2-20
Silicon dioxide 5-15
Hydroxypropyl Methylcellulose 2-10
Magnesium Stearate 2-10
Polyvinylpyrrolidone 5-10
Talc 1-10
Polysorbate 80 5-20
Sorbitol 5-20
Polyethylene glycol 1-10
IPA QS
Water QS
Average weight 400-460 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%.

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: 200-220 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 Biochanin-A 10.29 mg/kg 6
G4 Diacerein 10.29 mg/kg 6
G5 Bio-optimized PEA+ Biochanin-A 372 mg/kg+10.29 mg/kg 6
G6 Bio-optimized PEA+ Diacerein 372 mg/kg+10.29 mg/kg 6
G7 Biochanin-A + Diacerein 10.29 mg/kg +10.29 mg/kg 6
G8 Bio-optimized PEA+ Biochanin-A+ Diacerein
(RITANURON™) 372 mg/kg+10.29 mg/kg+10.29 mg/kg 6
G9 Pregabaline
(Lyrica® 75 mg) 20 mg/kg 6

Experimental Procedure:
Male Wister rat weighing between 200-220 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.

Result:
Table 2: Modulatory effect of test substances on Tail Immersion Latency reaction time (Seconds)
TAIL IMMERSION LATENCY REACTION TIME (Seconds)
Group
0 Min
(Sec) 30 Min
(Sec) 60 Min
(Sec) 90 Min
(Sec) 120 Min
(Sec) 180Min
(Sec)
G1 4.00±0.26 4.17±0.31 3.83±0.31 4.33±0.21 4.50±0.22 4.33±0.21
G2 4.17±0.17 8.50±0.22*
10.17±0.31*
10.83±0.31*
11.33±0.33*
11.50±0.22*

G3 4.00±0.26 7.33±0.42* 8.83±0.31* 9.50±0.22* 10.00±0.26* 10.17±0.31*
G4 3.83±0.31 7.17±0.31* 8.67±0.21* 9.50±0.22* 9.83±0.40* 10.00±0.26*
G5 4.17±0.17 8.50±0.22* 10.50±0.22* 11.33±0.33* 12.00±0.26* 12.67±0.21*
G6 4.33±0.21 8.33±0.21* 10.33±0.21* 11.17±0.31* 11.83±0.17* 12.50±0.22*
G7 4.33±0.21 8.17±0.31* 10.17±0.17* 11.00±0.26* 11.67±0.21* 12.33±0.21*
G8 4.50±0.22 9.00±0.26* 11.00±0.26* 12.00±0.26* 12.33±0.33* 13.00±0.26*
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*
*Values were expressed as Mean± SEM

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 -- 40.00 55.38 61.54 66.15 67.69
G3 -- 30.30 43.94 50.00 54.55 56.06
G4 -- 29.85 43.28 50.75 53.73 55.22
G5 -- 40.00 58.46 66.15 72.31 78.46
G6 -- 37.50 56.25 64.06 70.31 76.56
G7 -- 35.94 54.69 62.50 68.75 75.00
G8 -- 42.86 61.90 71.43 74.60 80.95
G9 -- 43.94 62.12 68.18 71.21 75.76

Discussion
In the present study analgesic activity was evaluated by tail immersion model. Rat were placed in to individual cylindrical mice holders leaving the tail hanging out freely. The animals were allowed to get acclimatized to mice 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± 0.5° 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.
Table - 03 shows the tail immersion latency reaction time after the administration of tests substance at different time interval. (G2), (G3), (G4), (G5), (G6), (G7), (G8), treated groups showed significant increased tail immersion latency reaction time when compared with Pain control group (G1).
(G2), (G3), (G4), (G5), (G6), (G7), (G8), and (G9) treated groups showed significant increased percentage maximum possible effect when compared with Pain control (G1) group at different intervals of time.

Conclusion: Overall result concluded that the combination of Bio-optimized PEA + Biochanin-A + Diacerein (G8) (80.95%), showed better percentage maximum possible effect at 180 min when compared with individual (G2) (67.69%), (G3) (56.06%), (G4) (55.22%) and marketed drug i.e., Lyrica® (G9) (75.76%).
,CLAIMS:We Claim:
1. A synergistic nutritional composition(s) for treating neuropathic pain, wherein the composition comprising a therapeutic blend of Palmitoylethanolamide (PEA), Biochanin-A and Diacerein along with pharmaceutically acceptable excipients, wherein the (PEA) and Biochanin-A and Diacerein are present in the ratio of 1: 0.001: 0.01 to 1: 0.02: 0.1 along with pharmaceutically acceptable excipients.

2. The synergistic nutritional composition as claimed in claim 1, wherein the PEA is present in the range of 50-98% by weight of total composition.

3. The synergistic nutritional composition as claimed in claim 1, wherein the PEA is in micronized form with improved bioavailability and solubility.

4. The synergistic nutritional composition as claimed in claim 1, wherein the Biochanin-A is present in the range of 0.1-2.0 % by weight of total composition.

5. The synergistic nutritional composition as claimed in claim 1, wherein the Diacerein is present in the range of 1.0 to 8.0 % by weight of total composition.

6. The synergistic nutritional compositions 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, solvent or mixtures thereof.

7. The synergistic nutritional compositions as claimed in claim 6, wherein the amount of diluent is present in the range of 1 to 40%; the amount of binder is present in the range of 0.2 to 30%; the amount of lubricant is present in the range of 0.1 to 5.0 %; the amount of glidant is present in the range of 0.1 to 5.0%; 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 5.0%, by weight of total composition.

8. The synergistic nutritional compositions as claimed in claim 1, wherein oral administration of an effective dose of the composition attenuates pain sensation and inflammation in a subject at early stage of neuropathic pain.

9. The synergistic nutritional compositions as claimed in claim 8, wherein the oral administration of the composition controls chronic pain.

10. The synergistic nutritional composition as claimed in claim 1, wherein the effective dose for daily oral administration is in the range of 300 to 600 mg.

11. A pain relieving synergistic nutritional composition comprising combination of Palmitoylethanolamide (PEA), Biochanin-A and Diacerein, wherein PEA and Biochanin-A and Diacerein are present in the ratio of 1: 0.001: 0.01 to 1: 0.02: 0.1 along with pharmaceutically acceptable excipients.

12. The pain relieving synergistic nutritional composition as claimed in claim 11, wherein the amount of PEA ranges from 50 to 98% by weight of total composition.

13. The pain relieving synergistic nutritional composition as claimed in claim 11, wherein the amount Biochanin-A ranges from 0.1 to 2.0 % by weight of total composition.

14. The pain relieving synergistic nutritional composition as claimed in claim 11, wherein the amount of Diacerein ranges from of 1.0 to 8.0% by weight of total composition.