DESC:Field of Invention
The present invention relates to nutraceutical composition for prevention and recovery from injury progression following spinal cord injury.
Background
Spinal cord injury is one of the most devastating traumatic conditions that induce primary and secondary tissue damage, resulting in postoperative complication, including spinal cord edema and cell death in the injured area.
The pathophysiology of SCI has two phases- primary and secondary phase. The primary injury serves as starting point from which secondary mechanism of injury extend; which involves cascades of cellular and biological events. Primary phase is the initial mechanical impact which directly leads to cell death and bleeding at impact site but it rarely fully disrupts the anatomical continuity of the spinal cord. Secondary phase starts with inflammation and haemorrhage within the gray matter leading to necrotic cell death or ischemia. The secondary phase sets in minutes after primary injury and can last for weeks to months. During this period the area of trauma distinctly enlarges and involves tissue destruction, which involves vascular dysfunction, edema, ischemia, excitotoxicity, electrolyte shifts, free radical production, inflammation, and strained apoptotic cell death [Tanhoffer et al. (2007) J Spinal cord Med. 30:140-146; Rowland et al. (2008). Neurosurg Focus 25:E2] Thus this period of secondary injury phase is a critical therapeutic target for the prevention of injury progression [Kim et al. (2017) Clin Orthop Surg 9:1-9].
The most immediate phenomenon of the secondary phase is neuro-inflammation. Pathological processes of neuro-inflammation include blood-spinal cord barrier (BSCB) damage. Structural damage to BSCB occurs almost at the moment of SCI, which usually leads to the increase of BSCB permeability. Thus, excess water, ions, inflammatory factors, immune cells (neutrophils, lymphocytes, etc) and foreign substances infiltrate into spinal cord parenchyma. It has also been reported that BSCB disruption is associated with increased mortality after endovascular therapy, and improvements in the BSCB functioning can significantly reduce secondary nerve injury. Therefore, prevention or recovery of the BSCB might improve outcome of SCI patients.
Another phenomenon of secondary phase of SCI which results in progression of the injury is characterized by increase production of reactive oxygen molecules (ROS) such as superoxide anions, hydrogen peroxide and peroxynitrite. Free radical-mediated oxidation of lipids, proteins and DNA injury, such as spinal cord hypoperfusion, development of oedema, axonal conduction failure and breakdown of energy metabolism. The importance of free radical mediated cell damage in the SIC is supported by the large number of experimental and clinical studies demonstrating the efficacy of antioxidants agents in improving neuronal damage. Inhibition or prevention of the free radical mediated damage to proteins, nucleic acids, lipids and extracellular matrix protein will prevent neuronal death.
The inventors of the present invention found that a multifactorial approach for preventing progression of SCI can be achieved by preventing or assisting in the recovery of BSCB and by inhibiting or decreasing the free radical mediated cell damage. Thus there is still need in the art to prepare a combination of active ingredient which will act synergistically to achieve aforementioned multifactorial approach for prevention of progression of SCI.
Multiple examples of use of ingredient for treatment of SCI are present in the art,
WO2019216380A1 discloses use of SSEA-3 (Stage-Specific Embryonic (Antigen-3) -positive pluripotent stem cells derived from living mesenchymal tissue or cultured mesenchymal cells for the treatment of spinal cord injury particularly include complete spinal cord injury or an incomplete spinal cord injury.
WO2007032396A1 discloses a therapeutic agent comprising a hepatocyte growth factor (HGF protein) or DNA encoding which are used for the treatment of spinal cord injury
US8481094B2 discloses method of treating neural injury e.g., spinal cord injury (SCI), or enhancing recovery from the neural injury with silymarin or silybin. Silymarin is natural extract obtained from the seeds of and fruits of the milk thistle Silybum marianum Gaertn. The prior art teaches silymarin and silybin are administered intrathecally via injection.
US10668032B2 discloses use of lithium benzoate for treating CND disorders. The prior art discloses lithium benzoate has potential therapeutic effect on spinal cord injuries.
But none of the existing art teaches any composition which will synergistically act to prevent the progression of SCI by preventing or assisting in the recovery of BSCB and by inhibiting or decreasing the free radical mediated cell damage. The inventors of the present invention surprisingly found that the same can be achieved by the use of a synergistic combination of polyphenol and Di-peptide. The polyphenol acts by assisting in the recovery of BSBC and preventing further deterioration of BSBC, whereas the dipeptide was found to act by inhibiting or decreasing the free radical mediated cell damage.
Objective
An objective of the present invention is the prevention of SCI progression, by assisting in the recovery of BSCB and by inhibiting or decreasing the free radical medicated cell damage.
Another objective of the present invention is to provide a composition for the prevention of SCI progression, by assisting in the recovery of BSCB and by inhibiting or decreasing the free radical medicated cell damage.
Summary
The present invention provides a synergistic nutraceutical composition comprising polyphenol and dipeptide to prevent second injuries resulting from spinal cord injury and process of preparation thereof.
In an another embodiment of the present invention provides an oral nutraceutical composition comprising polyphenol wherein the polyphenol is Curcumin and dipeptide is L- carnosine to prevent cell damage and inflammation associated with secondary injury resulting from spinal cord injury and assist in recovery of BSCB, wherein oral composition is a solid oral dosage form.
In an embodiment of the present invention provides a solid oral dosage form composition comprising curcumin and L- carnosine wherein solid oral dosage form is selected from tablet, capsule, pills, granules. In preferred embodiment the solid oral dosage form is capsule.
In an another embodiment the composition further comprises additional active agents wherein additional active agents are Vitamins and/or minerals.
Detailed Description
The present invention provides a nutraceutical composition comprising a fixed dose combination of effective amount of polyphenol and dipeptide to prevent or ameliorate secondary injuries resulting during SCI and process of preparation thereof.
In an embodiment the polyphenols are selected from but not limited to quercetin, kaempferol, catechins, stilbenes, curcumin or ellagic acid and/or combination thereof.
In a preferable embodiment the polyphenol is Curcumin. Curcumin is a polyphenol obtained from curcuma longa (turmeric), recently curcumin have been extensively studied for its role on spinal cord injuries. Nanwen Zhang et. al., studied the effect of curcumin injured spinal cord and found that curcumin inhibits secondary inflammation in the mice suffering from spinal cord injuries (SCI) and have neuroprotective effects on SCI.
In a preferred embodiment the nutraceutical composition of the present invention comprises curcumin in an amount ranging from 20 to 60%, most preferably 30% to 50% based on total weight of ingredients filled in the capsule.
In another embodiment the dipeptides are selected from carnosine, anserine, homoanserine, kyotorphin, balenine, pseudoproline, glycylglycine or combination thereof.
In a preferred embodiment the dipeptide is carnosine more preferably L-carnosine. L-carnosine is a dipeptide compound which is made up of two amino acids i.e. beta alanine and histidine. Carnosine is known to have antioxidant effect and have ability to suppress peroxidation and prevent free radical reactions; thus carnosine can be a potential anti-oxidant compound in eliminating reactive oxygen species (ROS) and oxidative stress, which play an important role in pathophysiology of SCI. [Quinn, P. J., et. al., Carnosine: Its properties, functions and potential therapeutic applications. Mol. Aspects Med. 13, 379–444.]
In an embodiment the nutraceutical composition of the present invention comprises L-carnosine present in amount ranging from 5-30%, more preferably 10-20% based on total weight of ingredients filled in the capsule.
Thus, in a preferred embodiment the nutraceutical composition of the present invention comprises 30% to 50% curcumin, 10-20% L-carnosine based on total weight of ingredients filled in the capsule.
In an another embodiment, the nutraceutical composition of the present invention is a solid dosage form selected from tablets, capsules, pills or granules. In a preferred embodiment, the solid dosage form is an oral capsule more preferably hard capsule
In an embodiment, the nutraceutical composition of the present invention provides a hard capsule comprising one or more pharmaceutical active agents and one or more pharmaceutical acceptable excipients. In another embodiment of the present invention, the pharmaceutical acceptable excipients are selected from binder, diluent, glidant and anti-caking agent or combinations thereof.
In another embodiment, the nutraceutical oral capsule comprises one or more active agents, pharmaceutical acceptable binder, diluent, glidant and anti-caking agent are in granules, pills or powder form. In most preferred embodiment, oral capsule comprises one or more active agents and pharmaceutical excipients in granule form.
In another embodiment, binders are selected from but not limited to starch, microcrystalline cellulose, lactose, guar gum, pectin, gelatine, polyvinyl pyrrolidone (PVP), sodium alginate or combination thereof.
In a preferred embodiment, binder is polyvinyl pyrrolidone (PVP) present in an amount ranging from 0.5-10%, more preferably 1-5% based on total weight of ingredients filled in the capsule.
In an another embodiment of the present invention, diluents are selected from but not limited to mannitol, lactose, starch, microcrystalline cellulose, calcium phosphate dihydrate, calcium phosphate trihydrate, dibasic calcium phosphate or combination thereof.
In a preferred embodiment of the present invention, diluent is microcrystalline cellulose present in an amount ranging from 10-50%, more preferably 20-40% based on total weight of ingredients filled in the capsule.
In an another embodiment of the present invention, glidants are selected from but not limited to talc, magnesium stearate, silicon dioxide, magnesium carbonate, silica or combination thereof.
In a preferred embodiment of the present invention, glidant is magnesium stearate present in an amount 0.5-10%, more preferably 0.5-5% based on total weight of ingredients filled in the capsule.
In a preferred embodiment of the present invention, anti-caking agent are selected from but not limited to magnesium stearate, calcium stearate, silica, talc, silicon dioxide, starch or combination thereof.
In a preferred embodiment of the present invention anti-caking agent is silicon dioxide present in an amount ranging from 0.5-10%, more preferably 0.5-5% based on total weight of ingredients filled in the capsule.
The hard capsule of the present invention can be manufactured by a method known to those skilled in the art, or a commercially available hard capsule can be also used. In an embodiment the hard capsule shell is made of hydroxypropyl methyl cellulose or gelatin more preferably hydroxypropyl methyl cellulose.
In an another embodiment of the present invention provides an oral nutraceutical composition in form of hard capsule comprising:
• Curcumin in an amount ranging from 30% to 50%.
• L-carnosine present in amount ranging from 10-20% based on total weight of ingredients filled in the capsule.
• Polyvinyl pyrrolidone (PVP) as a binder present in amount ranging from 1-5% based on total weight of ingredients filled in the capsule.
• Microcrystalline cellulose as diluent present in amount ranging from 20-40% based on total weight of ingredients filled in the capsule.
• Magnesium stearate as glidant present in an amount ranging from 0.5-5% based on total weight of ingredients filled in the capsule.
• Silicon dioxide as an anti-caking agent present in an amount ranging from 0.5-5% based on total weight of ingredients filled in the capsule.
In an another embodiment of the present invention, the composition comprises an additional active agent, wherein additional active agent is Vitamin D3. Oral Vitamin D3 helps in supplementing vitamin D3 deficiency in person suffering from SCI.
In a preferred embodiment, Vitamin D3 is present in an amount ranging from 0.05-5% more preferably 0.1-2% based on total weight of ingredients filled in the capsule with correspondence to the 200-800 IU per servings.
In an embodiment the nutraceutical composition of present invention ameliorates secondary injuries resulting from spinal cord injury by providing synergistic effect by-
i. Inhibiting or reducing secondary inflammation including blood-spinal cord barrier (BSCB) damage
ii. Preventing or inhibiting free radical mediated cell damage and oxidative stress.
In an another embodiment, the composition supplement vitamin D3 deficiency in a person suffering from spinal cord injury.
In an embodiment the present invention provides a hard capsule formulation comprises:
i. an effective amount of curcumin present in an amount ranging from 30% to 50% based on total weight of formulation and;
ii. an effective amount of l-carnosine present in an amount ranging from 10-20% based on total weight of formulation and;
iii. an effective amount of vitamin D3 present in an amount ranging from 0.05-5% more preferably 0.1-2% based on total weight of ingredients filled in the capsule with correspondence to the 200-800 IU per servings.
In a preferred embodiment, the present invention provides method of preparation of granules to be filled in hard capsule wherein the process involves wet or dry granulation.
The process of preparation is as follows:
1. L-carnosine and curcumin were dry mixed in rapid mixer granulator for 10 minutes and;
2. Binder solution was prepared by mixing Polyvinyl Pyrrolidone (PVPK 30) with isopropyl alcohol and;
3. The binder solution was then mixed with dry mixture obtained in step 1 in rapid mixer granulator to obtain wet granules and;
4. The wet granules were then dried in fluidised bed dryer and sized in multi-mill and;
5. The dried granules were mixed with Cholecalciferol and further lubricated with talc and magnesium stearate.
6. The granules obtained from step 5 were further filled in hard capsule.
The present invention is illustrated below by reference to the following examples. However, one skilled in the art will appreciate that the specific methods and results discussed are merely illustrative of the invention, and not to be construed as limiting the invention, as many variations thereof are possible without departing from the spirit and scope of the invention. The present invention is illustrated below by reference to the following examples. However, one skilled in the art will appreciate that the specific methods and results discussed are merely illustrative of the invention, and not to be construed as limiting the invention, as many variations thereof are possible without departing from the spirit and scope of the invention.
Example 1- Formula for oral nutraceutical composition:
Ingredients Wt.% per capsule Function
Vitamin D3 (Vitamin D3 500 IU/mg) 0.48 Active
Curcumin (Curcuma Longa Extract 7%) 44 Active
L- Carnosine 14.18 Active
Polyvinyl Pyrrolidone (PVPK 30) 2.84 Binding Agent
Iso Propyl Alcohol 1.70 Solvent
Microcrystalline Cellulose (MCC PH102) 34 Diluent
Magnesium Stearate 1.40 Glidant
Silicon Dioxide Amorphous 1.40 Anti-caking agent

Example-2: Process of preparation of present invention:
L-carnosine and curcumin were dry mixed in rapid mixer granulator for 10 minutes to obtain a dry mixture. The binder solution was prepared by mixing Polyvinyl Pyrrolidone (PVPK 30) with Alcohol. The binder solution was mixed with dry mixture comprising L-carnosine and curcumin in rapid mixer granulator to obtain wet granulation.
The wet granules were then dried in fixed bed dryer after dying the granules were sized in equal size. After sizing the granules were mixed with Cholecalciferol and were lubricated with magnesium stearate and talc. The granules were then filled in a hard capsule.

,CLAIMS:
1. A nutraceutical composition comprising synergistic combination of polyphenol and Di-peptide wherein nutraceutical composition is an oral solid dosage form.
2. An oral solid dosage form as claimed in claim 1 wherein polyphenol is Curcumin and dipeptide is L- carnosine or pharmaceutically acceptable salts thereof.
3. An oral solid dosage form as claimed in claim 2 wherein the formulation further comprises an additional active agent wherein additional active agent is vitamin D3.
4. An oral solid dosage form as claimed in claim 3 wherein solid oral dosage form is selected from tablet, capsule, pills, granules more preferably capsule.
5. A capsule formulation as claimed in claim 4 comprises:
iv. an effective amount of curcumin present in an amount ranging from 30% to 50% based on total weight of formulation and;
v. an effective amount of l-carnosine present in an amount ranging from 10-20% based on total weight of formulation and;
vi. an effective amount of vitamin D3 present in an amount ranging from 0.05-5% more preferably 0.1-2% based on total weight of ingredients filled in the capsule with correspondence to the 200-800 IU per servings.
vii. one or more pharmaceutically acceptable excipients.
6. A capsule formulation as claimed in claim 4 is a hard capsule wherein hard capsule shell is made of hydroxypropyl methyl cellulose (HPMC).
7. A hard capsule as claimed in claim 5 further comprises one or more excipients selected from binding agent, solvent, diluent, glidant and anti-caking agent.
8. A hard capsule as claimed in claim 6 comprises following excipients:
a) Polyvinyl pyrrolidone (PVP) as a binder present in amount ranging from 1-5% based on total weight of ingredients filled in the capsule and;
b) Microcrystalline cellulose as diluent present in amount ranging from 20-40% based on total weight of ingredients filled in the capsule and;
c) Magnesium stearate as glidant present in an amount ranging from 0.5-5% based on total weight of ingredients filled in the capsule and;
d) Silicon dioxide as an anti-caking agent present in an amount ranging from 0.5-5% based on total weight of ingredients filled in the capsule.
9. A hard capsule as claimed in previous claims wherein the capsule formulation ameliorates secondary injuries resulting from spinal cord injuries.
10. A process of preparation of hard capsule comprises following steps:
i. L-carnosine and curcumin were dry mixed in rapid mixer granulator for 10 minutes and;
ii. Binder solution was prepared by mixing Polyvinyl Pyrrolidone (PVPK 30) with isopropyl alcohol and;
iii. The binder solution was then mixed with dry mixture obtained in step 1 in rapid mixer granulator to obtain wet granules and;
iv. The wet granules were then dried in fluidised bed dryer and sized in multi-mill and;
v. The dried granules were mixed with Cholecalciferol and further lubricated with talc and magnesium stearate.
vi. The granules obtained from step 5 were further filled in hard capsule.

Dated this August 12, 2022
For Akums Drugs & Pharmaceuticals Ltd.

Siddhartha Dulakakhoria
(Authorized representative for Applicant)