FORM 2

THE PATENT ACT 1970

(39 of 1970)
&
The Patents Rules, 2003

COMPLETE SPECIFICATION

(See section 10 and rule 13)

1. TITLE OF THE INVENTION: ""NUTRITIONAL SUPPLEMENT" 2. APPLICANT
(a) NAME: CIPLA LTD.

(b)NATIONALITY: Indian Company incorporated under the Indian

Companies ACT, 1956
(c) ADDRESS: 289, Bellasis Road, Mumbai Central, Mumbai - 400 008,

Maharashtra, India
3.PREAMBLE TO THE DESCRIPTION
The following specification particularly describes the invention in which it is to be performed:

Technical field
The present invention relates to a nutritional supplement comprising green tea extract along with atleast one methylating agent and atleast one anti-oxidant or their pharmaceutically available salts, solvates, derivatives or enantiomers thereof in a pharmaceutically acceptable carrier. The invention further relates to pharmaceutical dosage forms and process thereof.
Background and prior art
Good nutrition is essential for optimal health; but finding time to eat a balanced diet on a daily basis seems a formidable task in this fast-paced, affluent society. Eating on the run and eating "fast-food" has become a part of our lifestyle leaving lots of holes in our seemingly healthy diets. In addition to this the body is constantly under attack from within as well as from external agents. Normal body functions such as breathing, physical activity, exposure to pollution and other lifestyle disorder such as stress produce substances called 'free radicals' that attack the healthy cells of the body. Hence the need for dietary supplements to help fight these free radicals
US6555142 describes a food supplement formulation comprising yi zhi ren, shan zhu yu, wu wei zi, ginseng, valerian root, passiflora incarnate, L-methionine, L-arginine, and a gender-specific complex selected from the group consisting of a mixture comprising beta-sitosteroi, saw palmetto, pollen extract, selenium, zinc, vitamin E, and lycopene, and a mixture comprising black cohosh, genistein, vitamin E, and cramp bark. More particularly, the invention is directed to a food supplement formulation which may additionally aid bladder control.
US2003133945 relates to a natural occurring food supplement for a human body's natural immune function, so as to be useful in the treatment of numerous medical problems, for the human being, specifically cancer.

WO 95/21542 Al discloses a beverage which is to be produced by the mixing of fruit juices and vegetable juices, oyster extract, glycyrrhizin, nonalcoholic red wine concentrate, wheat germ, soybean extract and seasonings. The beverage will contain provitamin A, vitamin A, vitamin E, vitamin C, trace elements, soluble fibers, flavonoids, glycyrrhizin, and taurine. It is also given in this citation that the red wine due to its flavonoid content has a preventative action against heart disease.
EP 0712630 A2 relates to an oral composition for prevention of sun allergies. The composition is based on a carotinoid, a tocopherol, ascorbic acid, and selenium. Carotene and lycopene are used as the carotinoids. EP 0712630 A2 mentions the already known combination of antioxidants with vegetable oil as the adjuvant in the production of capsules for oral use. EP 0 712 630 A mentions lecithin solely as the adjuvant for capsule production. Medical applications and/or effects besides the action against sun allergies of the oral administration of the preparation of EP 0712630 A2 are not disclosed.
WO03092413 relates to a food supplement which comprises folic acid, lycopene and an extract of Aspalathus linearis (common name Red Bush) or Cyclopia intermedia (common name Honeybush) or a mixture of extracts of Aspalathus linearis and Cyclopia intermedia.
Commercially many food supplements are available for help in and prevention of cardiac and circulatory conditions, relief as well as prevention of cancers, relief in rheumatic conditions, lipid-reducing action, antiarteriosclerotic action, strengthening of the immune system, action against premature aging processes, such as for example skin aging and/or organ aging.
Nevertheless there is still lacking a nutritional supplement that will fight the root cause of such disorders by regulating the metabolic pathways essential for normal growth, metabolism and repair the body and also protects the healthy cells from damage

Hence, the present invention aims to provide a novel, specifically designed combination of green tea extract along with atleast one methylating agent and atleast one anti-oxidant to protect against the progression of chronic ailments and degenerative disorders occurring due to micronutrient deficiency and free radical generation.
Object of Invention
An object of the present invention is to provide a nutritional supplement comprising green tea extract along with atleast one methylating agent and atleast one anti-oxidant or their phannaceutically available salts, solvates, derivatives or enantiomers thereof in a phannaceutically acceptable carrier.
The other object of the present invention is to provide a nutritional supplement that acts as a powerful disease retardant.
Another object of the present invention is to provide a method of preparation for the manufacture a nutritional supplement according to the present invention.
Another object of the present invention is to provide a nutritional supplement that has the ability to protect against the progression of chronic ailments and degenerative disorders occurring due to micronutrient deficiency and free radical generation.
Summary of Invention
According to the present invention there is provided a nutritional supplement comprising green tea extract along with atleast one methylating agent and atleast one anti-oxidant or their phannaceutically available salts, solvates, derivatives or enantiomers thereof in a phannaceutically acceptable carrier.

According to the second aspect of the present invention there is provided a nutritional supplement comprising antioxidants selected from the group comprising lycopene, selenium, ascorbic acid, vitamin E, P-carotene, lutein.
According to the third aspect of the present invention there is provided a nutritional supplement comprising methylating agents selected from the group comprising methionine, choline, folic acid Vitamin B6, Vitamin B12.
According to the fourth aspect of the present invention there is provided a nutritional supplement comprising methionine, lycopene and green tea extract.
According to the fifth aspect of the present invention there is provided a nutritional supplement comprising methionine, lycopene, choline and green tea extract.
According to the sixth aspect of the present invention there is provided a nutritional supplement comprising methionine, choline, folic acid, Vitamin B6, Vitamin B12, lycopene and green tea extract.
According to the seventh aspect of the present invention there is provided a nutritional supplement comprising green tea extract, methionine, choline, folic acid, Vitamin B6, Vitamin B12.
According to the eighth aspect of the present invention there is provided a nutritional supplement comprising green tea extract, vitamin E, ascorbic acid, selenium, lycopene, methionine, choline, folic acid, Vitamin B6, Vitamin B12.
According to the ninth aspect of the present invention there is provided a nutritional supplement comprising green tea extract, Vitamin E, ascorbic acid, selenium, lycopene, lutein, β- carotene, methionine, choline, folic acid Vitamin B6, Vitamin B12.

According to the tenth aspect of the present invention there is provided a process for the manufacture of a nutritional supplement comprising green tea extract along with atleast one antioxidant and atleast one methylating agent in a pharmaceutically acceptable carrier.
According to the eleventh aspect of the present invention there is provided a nutritional supplement comprising green tea extract along with atleast one antioxidants and atleast one methylating agents for retarding the progression of chronic diseases like hypertension, atherosclerotic cardiovascular diseases, cancer, diabetes, and neurological disorders and other disorders due to lack of nutrition.
Detailed Description
The need for a complete nutritional supplement to fight the root cause of diseases is evident from the above discussion. Hence, the present invention provides a novel, specifically designed combination of green tea extract along with atleast one methylating agent and atleast one anti-oxidant to protect against the progression of chronic ailments and degenerative disorders occurring due to micronutrient deficiency and free radical generation.
According to the present invention there is provided a nutritional supplement comprising green tea extract along with atleast one methylating agent and atleast one anti-oxidant or their pharmaceutically available salts, solvates, derivatives or enantiomers thereof in a pharmaceutically acceptable carrier.
Free radicals are electrochemically unbalanced molecules or atoms which have impaired electrons in their system. They acquire the missing electrons from their surroundings. This makes them highly aggressive chemicals, ready to react with cell components causing them permanent damage and leaving the body susceptible to cancer, heart disease, and many other degenerative diseases. It can also mutate cells and cause illness to the human being. Oxygen derived free radicals are the most reactive species which

attack the body. These toxic chemicals are formed as by-products during oxidative cellular metabolism.
Under physiological conditions, damage due to free radicals is countered by antioxidants. Antioxidant enzyme systems are a part of the body's immune system and nutrients that can protect us by stopping the free radical damage. The antioxidants neutralize free radicals by supplying the missing electron or removing an extra one to stabilize the molecule. The antioxidant then temporarily becomes a free radical but because of its structure it is not that damaging and it becomes stabilized through a chain reaction of other antioxidants.
According to the second aspect of the present invention there is provided a nutritional supplement comprising antioxidants selected from the group comprising lycopene, selenium, ascorbic acid, vitamin E, P-carotene, lutein.
Green tea (Camellia sinensis), an evergreen shrub native to eastern Asia, is cultivated extensively in India, China, Japan, Indonesia, Sri Lanka, Turkey, and Pakistan, as well as other parts of the world, such as Africa and South America. Green tea is prepared from the steamed and dried leaves of the shrub. Green tea contains polyphenols such as gallic acid and catechin, and their derivatives theogallin, gallocatechin, epiatechin, and epigallo catechin, epigallocatechin-3-gallate, epicatechin-3-gallate, other vitamins and minerals, tannins and flavonols. Tannin is a nutrient antioxidant and catechin is a powerful antioxidant. The antioxidants help our body to fight against free radicals. Catechins are powerful antioxidants that fight viruses, slow aging, and have a beneficial effect on health. Catechins also destroy free radicals, and have far-reaching positive effects on the entire body. One major element in Green tea is Epigallo Catechin - 3 - gallate (EGCG), a compound that is believed to block production of an enzyme required for cancer cell growth. EGCG works by suppressing the formation of blood vessels, a process called angiogenesis, thereby cutting off the supply of blood to cancer cells.
Other pharmacological actions of green tea include effects on lipid metabolism; antimicrobial, antioxidant, and antimutagenic effects; and a wide variety of anticancer

actions. It may also have thermogenic and antiinflamatory properties. It also reduces high blood pressure, lowers blood sugar, helps reduce body fat.
Generally the extract of green tea is used instead of using the leaves. Green Tea Extract can be obtained by different processes but preferably the aqueous extract is used. The aqueous extract can be produced by various processing steps, including filtration, concentration and drying.
The green tea extract can be of different grades depending on the content of polyphenols namely catechins, epigallocatechin gallate (EGCG) and caffeine. The extract may be available as comprising between 25% polyphenols and 98% polyphenols which comprise EGCG in a range between 12% and 55%, catechins in a range between 12% and 85%, caffeine in a range between 0% to 10%. The supplement may comprise green tea extract preferably between 125 and 1000mg
Lycopene is a carotenoid from the family of beta carotenes. Lycopene is one of the approximately 600 known carotenoids, chemicals that are responsible for the red, orange, and yellow colour in fruits and vegetables. Lycopene is found primarily in tomatoes and is known to have potent antioxidant capabilities and remarkable fighting power against degenerative diseases. Lycopene has a single oxygen quenching ability as high as that of beta-carotene and 10 times higher than that of alpha-tocopherol. Lycopene has the ability to donate electrons to quench and neutralize free radical oxygen molecules before they damage the cells. It is therefore known as a biological quencher of free radicals especially those derived from oxygen.
Lycopene acts as an antioxidant, may trap the oxygen species, increase the overall antioxidant potential or reduce the oxidative damage to lipids (lipoproteins, membrane lipids), proteins (important enzymes) and DNA (genetic material), thereby lowering oxidative stress. This reduced oxidative stress may lead to reduced risk of cancer and cardiovascular disease.

Increased Lycopene status may regulate gene functions, improve intracellular communication, modulate hormone and immune response, or regulate metabolism, thus lowering the risk for chronic disease. These mechanisms may also be interrelated and may operate simultaneously to provide health benefits. The antioxidant activity of lycopene may be additive or possibly synergistic with that of ascorbic acid and vitamin E
Lycopene has a role in preventing carcinogenesis and atherogenesis by protecting critical cellular biomolecules including lipids, lipoproteins , proteins and DNA. The supplement may comprise lycopene preferably between 0.5mg and 25mg
The Lycopene is available in different forms and can be suitably used as per requirement for desired formulation. It may be used as a 5% powder or as a 6% dispersion in oil. The oil may be any pharmaceutically acceptable oil. The oil used may be canola oil. Canola oil has a better fatty acid ratio as compared to other oils and is favorable in terms of health benefits and nutritionally balanced diet, and hence the preferred oil is canola oil. It has high percentage of monounsaturated fatty acid, oleic acid about 61%. Oleic acid has shown to reduce serum cholesterol levels and LDL cholesterol levels. Oleic acid does not affect levels of HDL cholesterol.
Vitamin E is a fat-soluble vitamin existing in eight different forms, of which alpha tocopherol is the most active form of vitamin E in humans, and is a powerful biological antioxidant. Alpha tocopherol is available in different forms of which the one desired can be used. Alpha tocopheryl acetate is an acetate ester of natural source of alpha tocopherol and a powerful antioxidant and may be preferred to formulate supplements. Vitamin E protects against damaging effects of free radicals, which may contribute to the development of chronic diseases such as cancer. Vitamin E may block the formation of nitrosamines, which are carcinogens formed in the stomach from nitrites consumed in the diet. It may also protect against the development of cancers by enhancing the immune function. Vitamin E may help prevent or delay coronary heart disease. Oxidative modification of LDL-cholesterol promotes blockages in coronary arteries that may lead

to atherosclerosis and heart attacks. Vitamin E helps prevent or delay coronary heart disease by limiting oxidation of LDL-cholesterol
Vitamin E may also have antithrombotic, anticoagulant, neuroprotective, antiproliferative, immunomodulatory, cell membrane stabilizing and antiviral actions. The supplement may comprise vitamin E preferably between 5 mg and 250mg
Preferably the Alpha-Tocopherol acetate can be used as a concentrate (powder form) which can be prepared either by finely dispersing alpha-Tocopherol acetate in a suitable carrier of suitable quality (for example gelatin, acacia, carbohydrates, lactoproteins or a mixture thereof) or by adsorbing alpha -Tocopherol acetate on silicic acid of suitable quality. It can be advantageously gelatin coated.
Selenium is an essential trace element in human nutrition and is involved in the defence against the toxicity of reactive oxygen species, in the regulation of thyroid hormone metabolism and the regulation of redox state of cells. Selenium is a part of the enzyme glutathione peroxidase, which metabolizes hydroperoxides formed from polyunsaturated fatty acids. Selenium is also a part of enzymes that deiodinate thyroid hormones. Further selenium is essential for normal functioning of the immune system and thyroid gland. Deficiencies of selenium have been associated with an increased cancer risk. Improved selenium nutrition may reduce the incidence of several kinds of cancer. The protective effects of selenium seem to be primarily associated with its presence in the glutathione peroxidases which are known to protect DNA and other cellular components from damage by oxygen radicals. Selenoenzymes are also known to play roles in carcinogen metabolism, in the control of cell division, oxygen metabolism, detoxification processes , apoptosis induction and the functioning of the immune system. This nutrient is an important part of antioxidant enzymes that protect cells against the effects of free radicals that are produced during normal oxygen metabolism. Selenium limits the oxidation of LDL cholesterol and thereby helps to prevent coronary artery disease.

Generally, selenium functions as an antioxidant that works in conjunction with vitamin E
Selenium is also required for the proper functioning of the immune system and appears to be a key nutrient in counteracting the development of virulence and inhibiting HIV progression to AIDS.
Selenium has antioxidant activity and may also have immunomodulatory activity. It may also have activity in detoxification of some metals and other xenobiotics and activity in fertility enhancement in males. The supplement may comprise selenium preferably between 0.02 and 0.25mg
Ascorbic acid is generally known as Vitamin C which is a water-soluble vitamin. Humans depend on ascorbic acid for many aspects of our biochemical functioning; yet we are among only a handful of animal species that cannot produce their own supply of vitamin C.
Vitamin C plays a major role in the manufacture and defense of our connective tissue, the elaborate matrix that holds the body together. It serves as a primary ingredient of collagen. Vitamin C enhances the immune system and supports the cardiovascular system by facilitating fat metabolism and protecting tissues from free radical damage. Vitamin C assists the nervous system by converting certain amino acids into neurotransmitters. As an antioxidant, primary role of vitamin C is to neutralize free radicals. Since ascorbic acid is water soluble, it can work on both, inside and outside the cells to combat free radical damage.
Vitamin C reduces the oxidative stress associated with chronic disease like cancer. Vitamin C may prevent this plaque formation by inhibiting the oxidative modification of low-density lipoproteins (LDLs). Vitamin C is found to significantly reduce the lipid peroxide levels and oxidative load in coronary heart disease patients. Vitamin C can enhance the body's resistance to an assortment of diseases, including infectious disorders and many types of cancer. It strengthens and protects the immune system by stimulating the activity of antibodies and immune system cells such as phagocytes and neutrophils.

Vitamin C works along with vitamin E and the enzyme glutathione peroxidase to stop
free radical chain reactions.
Vitamain C may also have antihypersensitive, antiviral , antihistaminic,
immunomodulatory, opthalmoprotective and airway protective actions. It may aid in the
detoxification of some heavy metals, such as lead and other toxic chemicals.
The supplement may comprise ascorbic acid between 50mg and 1000mg.
P-carotene is a member of a class of substances called carotenoids. Carotenoids are
principle pigments responsible for the red, orange, yellow, and green colors of vegetables
and fruits and is also responsible for the color of carrots. It has antioxidant activity and
may also have immunomodulatory, anticarcinogenic and antiatherogenic activities in
some cases. It quenches singlet oxygen, scavenges peroxyl radicals and inhibits lipid
peroxidation.
The supplement may comprise β-carotene preferably between 3mg and 15mg
Lutein is a member of the carotenoid family and appears to protect humans from phototoxic damage. It is found in the macula of the human retina as well as human crystalline lens. It is thought to play a role in protection against age related macular degeneration and age related cataract formation. The possible protection afforded by it may be accounted for in part by its reactive oxygen species scavenging ability. The supplement may comprise lutein preferably between 0.25mg and 20mg of lutein.
According to the third embodiment of the present invention there is provided a nutritional supplement comprising methylating agents selected from the group comprising methionine, choline, folic acid Vitamin B6; Vitamin B12.
Restoring SAM (S- Adenosyl Methionine) is the key to enhancing methylation. Many studies point to free radicals as a major cause of degenerative disease and free radicals are neutralized by anti-oxidants. The high prevalence of degenerative diseases such as cancer and atherosclerotic vascular diseases suggest that the so-called healthy diet is not

sufficient in terms of anti-oxidants. Methylation also affects free radical status, and vice versa. Enhancing one without the other is like a job half done.
Methylation is a critical biochemical reaction involving transfer of a "methyl group (-CH3) " to another molecule. This attachment of methyl group to different molecules is involved in dozens of important life functions such as synthesis and repair of DNA, synthesis of proteins, metabolism of lipids, maintaining an optimal immune system and controlling gene function.
Methylation is a biochemical process that starts with methionine. When methionine reacts with ATP(Adenosine triphosphate), S- Adenosyl Methionine (SAMe) is formed. SAMe contains the "methyl groups" (-CH3) necessary for methylation. In methylation, these groups break off from SAMe to catalyze other reactions. The leftover product eventually becomes homocysteine. Normally, homocysteine is converted to cysteine, or back to methionine in the presende of methylating agents. If not converted, homocysteine can inhibit methylation.
Folic acid, vitamin B12, vitamin B6, Methionine and choline all are important for optimum methylation. Deficiency of these methylating agents can lead to sub- optimal methylation. Folic acid, vitamin B6, vitamin B12 and choline are required for metabolism of homocysteine. Nutritional deficiencies of these agents or genetic defects of some enzymes in the homocysteine metabolism can lead to an increase in homocysteine plasma concentration and is associated to an increment in atherosclerotic vascular diseases. Methylation keeps check on cancer genes. Sub-optimal methylation can lead to development and progression of cancer.
Methionine is an essential amino acid that is not synthesized by the body and must be obtained from food. It is one of the sulphur containing amino acids and is important in many body functions. Through its supply of sulphur, it improves the tone and pliability of the skin, conditions the hair and strengthens nails. The mineral sulphur also protects the cells from airborne pollutants, such as smog, slows down the aging process in the cells,

and is involved with the production of protein. It also acts as a lipotropic agent to prevent excess fat buildup in the liver. S-Adenosyl Methionine, an amino acid, is manufactured from methionine. SAMe is the most active methyl-donor in the body, i.e. it donates methyl groups to other chemical compounds in the body and also plays role in DNA methylation. Methionine helps to eliminate fatty substances that obstruct arteries, including those that supply the brain, heart, kidneys. SAMe donates methyl groups to chemical compounds including neurotransmitters, changing them into other compounds. Supplementation with SAMe increases levels of serotonin and dopamine and improves serotonin and dopamine receptor site binding.
Methionine has anticarcinogenic, antiatherosclerotic action and also helps prevent neuorodegenerative disease. It is a excellent chelator of heavy metals and aid in their excretion from the body.Methionine is essential for the absorption and transportation and bioavailabilityof selenium and zinc in the body The supplement may comprise methionine preferably between 20mg and 1000mg.
According to the fourth aspect of the present invention there is provided a nutritional supplement comprising methionine, lycopene and green tea extract.
Choline is an essential nutrient. Although, humans can synthesize it in small amounts, choline must be consumed in the diet to maintain health. The three major metabolic functions of choline are as a precursor for phosphatidylcholine biosynthesis, as a precursor for acetylcholine biosynthesis and as a methyl donor. The majority of the body's choline is found in specialized fat molecules known as phospholipids, the most common of which is called phosphatidylcholine or lecithin. Choline and its metabolites serve a number of vital biological functions.
Choline is vital for maintaining structural integrity of cell membranes. It is used in the synthesis of phospholipids, phosphatidylcholine and sphingomyelin - the structural components of all human cell membranes. Choline is a pecursor for acetylcholine, an important neurotransmitter involved in muscle control, memory and many other

functions.Phosphatidylcholine is a required component of very low density lipoproteins
which are required for the transport of fat and cholesterol from liver through blood tissues
that require them. Without adequate phosphatidylcholine, fat and cholesterol accumulate
in the liver. Choine is important for normal cellular membrane composition and repair,
normal brain function and normal cardiovascular function. Choline also plays role in
decreasing the risk of cancer, cardiovascular diseases, pregnancy complications and
alzheimer's disease.
Choline can be used as its various available salts.Preferably the bitartarate salt may be
used..
The supplement may comprise choline preferably between 2mg and 50mg.
According to the fifth aspect of the present invention there is provided a nutritional supplement comprising methionine, choline, lycopene and green tea extract.
Folic acid is an essential, water soluble B-vitamin found in all living organisms. It is chemically unstable and is lost during the preparation of food. Folic acid is required for cell replication and growth. Folic acid is required for synthesis of DNA. Folic acid is needed for protein synthesis in all cells. Therefore rapidly growing tissue, such as the foetus, and rapidly repaired cells like red blood cells and immune cells have a high need for folic acid. Folic acid is also required for the metabolism of homocysteine. Folic acid is necessary not only for the synthesis of DNA, but also to maintain the integrity of DNA. Mutations and breaks in DNA strands can transform a normal cell into cancerous cell. High intake of folate has beneficial role in the prevention of colorectal and breast cancer. Higher intakes of folate lower the risk of colorectal adenoma and cancer. Folic acid supplements may help reduce hyperhomocysteinaemia. The lowering of lipid peroxidation due to reduction in plasma homocysteine levels reduce the risk of atherosclerosis and prevents or delays cardiovascular complications. Folic acid lowers the risk of neural tube defects and possibly other types of birth defects .It may also have neuroprotective and antidepressant actions. The supplement may comprise folic acid preferably between 0.2 and 1.5mg.

Vitamin B6 or pyridoxine is a water-soluble vitamin. There are six forms of vitamin B6 of which pyridoxal 5' phosphate (PLP) is the active coenzyme form, and has the most importance in human metabolism. PLP plays a vital role in the function of approximately 100 enzymes that catalyze essential chemical reactions in the human body. In red blood cell formation and function,Vitamin B6 also known as pyridoxine hydrochloride functions as a coenzyme in the synthesis of heme, a component of haemoglobin. In Nucleic acid synthesis, Vitamin B6 serves as a coenzyme for the synthesis of nucleic acids (DNA and RNA). In immune function, Low Vitamin B6 intake and status have been associated with impaired immune function due to decreased production of immune system cells known aslymphocytes and interleukin-2. In nervous system function,the synthesis of neutrotransmitter, serotonin, from the amino acid tryptophan, in the brain is catalysed by a VitaminB6 dependent enzyme. Other neurotansmitters such as dopamine, norepinephrine and gamma-aminobutyric acid are also synthesized using VitaminB6 dependent enzymes. Vitamin B6 dependent enzymes are required for metabolism of homocysteine to cysteine. Low Vitamin B6 intake leads to increased blood homocysteine levels and increased risk of cardiovascular diseases. Vitamin B6 supplementation has been found effective in lowering blood homocysteine levels. Decline in cognitive function for alzheimer's disease in the elderly is associated with inadequate nutritional status of folic acid, vitamin B6 and vitamin B12. Higher plasma levels of vitaminB6 have been associated with better performance in cognitive function.
Vitamin B6 has antineurotoxic activity and may have activity in a number of inborn
metabolism. It has putative antiatherogenic, anticarcinogenic and mood modulatory
activities.
The supplement may comprise vitamin B6 preferably between 0.5mg and 50mg.
Vitamin B12 also known as cyanocobalamin is effective in the treatment of pernicious anaemia. Absorption of vitamin B12 from foods is complex and a defect in any step can lead to deficiency.

Two cyanocobalamin dependent enzymatic reactions occur in humans. The first reaction converts methylmalonyl-coenzyme A (CoA) to succinyl-CoA using cobalamin as a cofactor. Vitamin B12 deficiency leads to an increase in serum methylmalonyl-coenzyme A and its metabolic product, methylmalonic acid (MMA). The second reaction uses cobalamin as a cofactor in the synthesis of methionine from homocysteine. Deficiency in B12 leads to an accumulation of homocysteine. DNA damage from micronutrient deficiencies is likely to be a major cause of cancer. Vitamin B12 deficiency may cause high uracil and chromosome breaks. Homocysteine lowering therapy with folic acid, vitamin B6 and vitamin B12 significantly reduces the incidence of major adverse events after percutaneous coronary intervention.
Vitamin B12 is used in the treatment of megaloblastic anaemia.it may have antiatherogenic, neuroprotective, anticarcinogenic and detoxifying activities. It has putative antiallergic and mood modulatory acivities. The supplement may comprise vitamin B12 preferably between 0.0l mg and 0.03mg
Thus the present invention is scientifically designed combination of the right methylating agents and the right antioxidants to reduce the risk of chronic diseases and degenerative disorders occurring due to micronutrient deficiency and free radical generation. The methylating agents maintain optimum methylation, whereas the green tea and the antioxidants reduce the oxidative stress associated with chronic diseases like cancer, atherosclerosis, diabetes etc.
According to the sixth aspect of the present invention there is provided a nutritional supplement comprising methionine, choline, folic acid Vitamin B6, Vitamin B12, lycopene and green tea extract.
According to the seventh aspect of the present invention there is provided a nutritional supplement comprising green tea extract, methionine, choline, folic acid, Vitamin B6 Vitamin B12.

Vitamin B12 is highly unstable and light sensitive. It is required in low doses and are also oily in nature and thus gives problem with respect to content uniformity. It is therefore diluted with diluents known in the art like starch, lactose. Alternatively it is gelatin coated whereby it gets diluted as well as stabilized. The alpha tocopheryl acetate, is also oily in nature and also required in low doses and hence would pose a problem in content uniformity, and hence it is diluted. Preferably the alpha tocopheryl acetate and Vitamin B12 are gelatin coated.
According to the eight aspect of the present invention there is provided a nutritional supplement comprising green tea extract, vitamin E, ascorbic acid, selenium, lycopene, methionine, choline, folic acid, Vitamin B6; Vitamin B12.
According to the ninth aspect of the present invention there is provided a nutritional supplement comprising green tea extract, vitamin E, ascorbic acid, selenium, lycopene, lutein, β- carotene, methionine, choline, folic acid Vitamin B6, Vitamin B12.
The active ingredients in the class of "methylating agents, antioxidants" are used in the entire specification and claims in a broad sense to include not only the active ingredients in the class of "methylating agents, antioxidants" per say but also their pharmaceutically available salts,solvates, derivatives or enantiomers thereof
It is preferred that the defined constituents be in a pharmaceutically acceptable dosage form. The dosage form can be in the form of tablets, hard gelatin capsules, soft gelatin capsules, paste, beads, dry syrup, sachet, rediuse, liquid orals, powder, pills, syrup, pharmaceutically acceptable oil, a solution or a suspension or similar conventional dosage forms, using conventional equipment and techniques known in the art. However tablets forms are preferred. When constituents are in oil, the oil can be within a soft gel capsule or a hard shell capsule.
According to the tenth aspect of the present invention there is provided a process for the manufacture of a nutritional supplement comprising green tea extract along with atleast

one methylating agent and atleast one anti-oxidant in a pharmaceutically acceptable carrier. The process may involve conventional techniques known in the art for the formulation of different dosage forms.
In solid dosage forms of the invention for oral administration (capsules, tablets, pills, powders and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, and/or any of the following:
Diluents comprising one or more of but not limited to dibasic calcium phosphate, magnesium oxide, light magnesim oxide, calcium carbonate, starch, starch 1500, maltose, calcium sulphate, magnesium carbonate, cellulose-microcrystalline, cellulose powdered, dextrates, dextrins, dextrose, fructose, lactose, mannitol, sorbitol, starch, starch pregelatinized, sucrose, sugar compressible, sugar confectioners and equivalents thereof. The diluent/s can preferably be used in a range of 10-90% w/w;
Binders comprising one or more of but not limited to methyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose (HPMC), polyvinylpyrrolidone, gelatin, gum arabic, ethyl cellulose, polyvinyl alcohol, pullulan, pregelatinized starch, tragacanth, sodium alginate, propylene glycol and alginate, and in particular may be gelatin, hydroxy ethyl cellulose , methyl acrylates dispersion, methyl cellulose, veegum, xanthan gum, maltodextrin. The binder/s can preferably be used in a range of 2-20% w/w. Disintegrants comprising one or more of but not limited to hydroxypropyl cellulose, prosolv, carboxymethylcellulose, microcrystalline cellulose, calcium carboxymethylcellulose, crospovidone, sodium starch glycollate, sodium carboxymethylcellulose, croscarmellose sodium, starch, crystalline cellulose, hydroxypropyl starch, partly pregelatinized starch and equivalents thereof. The disintegrant/s can preferably be used in a range of 5-30% w/w; Lubricants/glidants comprising one or more of but not limited to stearic acid, magnesium stearate, calcium stearate, talc, hydrogenated caster oil, colloidal silicon dioxide, sucrose esters of fatty acid, microcrystalline wax, yellow beeswax, white beeswax, silicon dioxide and equivalents thereof.The lubricant(s)/glidants(s) can preferably be used in a range of 0.25-3%w/w.

Optionally they may also comprise absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, but not limited to cetyl alcohol and glycerol monostearate; absorbents, such as but not limited to Colloidal silicon dioxide, cellulose powder and coloring agents. In the case of capsules, tablets, pills and powders, the therapeutic formulations may also comprise buffering agents. Solid formulations of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
Tablets may be made by compression or molding powdered components, optionally with one or more pharmaceutically acceptable carriers. Compressed tablets may be prepared using at least one of binder, lubricant, inert diluent, preservative, disintegrant), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered component moistened with an inert liquid diluent. In one preferred aspect green tea extract, methionine & choline may be put into lycopene dispersion on oil, and further granulated with selenium, ascorbic acid and one or more diluents and/or binders to form granules; further the granules can be mixed with folic acid, vitamin B6, vitamin B12 vitamin E and one or more pharmaceutically acceptable excipients to form a mixture; this may then either be encapsulated or compressed to form a tablet.
In another preferred aspect the methionine, choline and green tea extract may be granulated using suitable binder, selenium, vitamin B6 and ascorbic acid may be mixed and granulated using suitable binder. Granules from both the above steps and the remaining active ingredients can be blended with lubricants. This may then either be encapsulated or compressed to form a tablet.
The tablets, and other solid dosage forms of the therapeutic formulations of the present invention, such as capsules, powders and/or pills, may optionally be prepared with coatings and shells, such as coatings well known in the art. The coating can be any conventional coating . Preferably the coating may be film coating. The coating composition may contain all conventional film forming materials like but not limited to

cellulosic polymers such as xanthan gum, polyvinyl alcohol Hydroxypropyl methylcellulose, ethyl cellulose, hydroxy ethyl cellulose and the like, polyvinyl polymers such as polyvinyl acetate, polyvinyl pyrrolidone, polyvinylacetophthalate and the like, acrylates such as eudragit-E etc., plasticisers such as but not limited to propylene glycol, polyethylene glycol, triacetine, triethyl citrate, dibutyl sebecate, polysorbates. Opacifiers such as but not limited to titanium dioxide, light magnesium oxide, magnesium oxide , talc , coloring agents such as but not limited to water soluble and water insoluble dyes, drying agents such as starch, talc, magnesium oxide. The polymers may be used in vehicles such as but not limited to water, isopropyl alcohol, methylene chloride, ethanol, propylene glycol, dichloromethane etc or mixtures thereof Alternatively film coating redimix, color redimix may also be used. They may optionally be coated with a seal coat prior to the film coating using conventional seal coating agents known in the art. The coated formulation may further be polished using pharmaceutically acceptable waxes. The optional wax polish includes a waxy material. Suitable waxy materials include one or more of but not limited to shellac, modified shellac (opaglos), Opaglos II, carnuba wax, bees wax, paraffin wax, polyethylene glycol, microcrystalline wax, stearic acid and the like. One application is usually sufficient to obtain the desired effect.
Liquid form preparations include solutions, suspensions, and emulsions and the like. Aqueous solutions suitable for oral use are prepared by dissolving the active component in water or other suitable liquid and adding one or more of suitable colorants; flavors; preservative agents such as but not limited to such as methylparaben, propylparaben, and sodium benzoate, benzoic acids; stabilizing agents; surfaceactive agent such as but not limited to such as sodium lauryl sulfate, polysorbate 80, lutrols, polyethylene glycols; thickening agents; sweeteners; Buffering agents as desired and the like. Aqueous solutions suitable for oral use may also be made by dispersing the finely divided active component in water or other suitable liquid with viscocity modifying materials, such as but not limited to natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other suspending agents known in the art such as but not limited to microcrystalline cellulose, hydroxypropyl methyl cellulose, xanthan gum, sodium carboxymethyl cellulose , povidone, polyvinyl alcohol, colloidal silicon dioxide

and the like. The liquid formulations may also comprise vehicles such as but not limited to aqueous sorbitol, glycerin, sugar syrup, propylene glycol, polyethylene glycols, maltitol, oily vehicle bases such as but not limited to sunflower oil, arachis oil, soya oil etc; Chelating agents such as but not limited to dissolved EDTA, citric acid, Tartaric acid etc; antioxidants such as but not limited to BHA, BHT, tocopherol acetate, propyl gallate etc
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions, and emulsions. These particular solid form preparations are provided in unit dose form and as such are used to provide a single liquid dosage unit. Alternatively, sufficient solid preparation may be provided so that the after conversion to liquid form, multiple individual liquid doses may be obtained by measuring predetermined volumes of the liquid form preparation as with a, teaspoon, or other volumetric container.
The solid and liquid forms may contain, in addition to the active material, one or more of flavorants; colorants; stabilizers; buffers; artificial and natural sweeteners; dispersants; thickeners; solubilizing agents; glidants like but not limited to colloidal silicon dioxide and the like. The liquid utilized for preparing the liquid form preparation is suitably water, isotonic water, ethanol, glycerin, propylene glycol, and the like, as well as combinations thereof. The liquid utilized will be chosen with regard to the route of administration
According to the eleventh aspect of the present invention there is provided the use of the nutritional supplement comprising atleast one antioxidants and atleast one methylating agents for retarding the progression of chronic diseases like hypertension, atherosclerotic cardiovascular diseases, cancer, diabetes, and neurological disorders.
It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the spirit of the invention. Thus, it should be understood that although the present invention

has been specifically disclosed by the preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and such modifications and variations are considered to be falling within the scope of the invention.
The following examples are for the purpose of illustration of the invention only and are not intended in any way to limit the scope of the present invention.
Example 1: Nutritional supplement as a tablet

Sr. No. Ingredients Mg/tab
1. Lycopene (6% Dispersion in canola oil) 6.00
2. Selenious acid equivalent to selenium 0.05
3. DL-Methionine 50.00
4. Choline Bitartarate Equivalent toCholine 10.00
5. Green Tea extract 250.00
6. Ascorbic Acid (Vitamin C) 300.00
7. Light Magnesium oxide 80.00
8. Colloidal silicon dioxide 20.00
9. Microcrystalline cellulose 100.00
10. Gelatin 28.0
11. Purified water q.s
12. Folic acid 0.075
13. Pyridoxine Hydrochloride (vitamin Be) 24.00
14. Cyanocobalamin (Vitamin B12) 0.02
15. Alpha Tocopheryl Acetate Concentrate 19.46
16. Microcrystalline cellulose 281.023
17. Mannitol 30.0
18. Crospovidone 125.00
Seal coating
19. Hydroxypropylmethylcellulose 15.40
20. Ethyl cellulose 5.10
21. Talc 15.40

22. Light Magnesium oxide 10.30
23. Propylene Glycol 3.80
24. Isopropyl alcohol -
25. Dichloromethane -
Colour Coating
26. Hydroxy propyl methyl cellulose 15.95
27. Caramel 12.00
28. Colour Natural Caramel 16.50
29. Talc 4.85
30. Propylene Glycol 2.20
31. Purified Water -
Wax coating
32. Opaglos 6000 1.50
33. Isopropyl alcohol -
l.Selenious acid, Choline Bitartrate and DL Methionine were dispersed in Lycopene.
2. The above dispersion was adsorbed over Light Magnesium oxide and Colloidal
silicon dioxide and granulated with the binder.
3. These granules and the remaining active ingredients were blended with
lubricants. The blend so formed was compressed into tablets using suitable
toolings.
4. The tablets were further film coated with HPMC based coating solutions and
polished with wax coating.
Example 2:Nutritional supplement as a capsule

Sr. No. Ingredients Mg/cap
1. Lycopene (6% Dispersion in canola oil) 3.00
2. Selenious acid equivalent to selenium 0.025
3. DL-Methionine 25.00
4. Choline Bitartarate Equivalent toCholine 5.00
5. Green Tea extract 125.00
6. Ascorbic Acid (Vitamin C) 150.00
7. Light Magnesium oxide 40.00

1.Dispersion of Selenious acid, Choline Bitartrate and DL Methionine in Lycopene was prepared.
2. This dispersion was adsorbed over Light Magnesium oxide and Colloidal silicon dioxide and granulated with the binder.
3. The above granules and the remaining active ingredients were blended with lubricants.
4. The blend was then filled in hard gelatin capsules of suitable size.
Example 3:Nutritional supplement as a dry syrup



1. Actives were mixed with all excipients, buffers, sweetners, flavours, viscosity modifiers, to form premix in a suitable blender.
2. The above premix was diluted in geometric proportion with the diluent i.e. sugar & finally blended in a suitable blender.
3. This blend was then filled in dry syrup bottles as per the proposed fill weights.
Example 4: Nutritional supplement for rediuse



1. Actives were mixed in a suitable blender.
2. Preservatives methyl paraben & propyl paraben were dissolved in oil by slow heating.
3. To above antioxidants BHA, Anti-adherent, colloidal silicon dioxide & sweetner -sodium
saccharin were added to dissolve.
4.Active premix was added & volume mode up with Oil.
5. Final dispersion was filtered & then filled in bottles.

Example 5



1. Granules of DL Methionine, Choline Bitartarate and Green Tea Extract were prepared with suitable binder.
2. Selenious acid, Pyridoxine Hydrochloride and Ascorbic acid were mixed and granulated with suitable binder

3. Granules from both the above steps and the remaining active ingredients were blended with lubricants. The blend so formed was compressed into tablets using suitable toolings
4. The tablets were further film coated with HPMC based coating solutions and polished with wax coating

We claim,
1) A nutritional supplement comprising green tea extract along with atleast one methylating agent and atleast one anti-oxidant or their pharmaceutically available salts, solvates, derivatives or enantiomers thereof in a pharmaceutically acceptable carrier
2) A nutritional supplement as claimed in claim 1, wherein the antioxidants are selected from the group comprising lycopene, selenium, ascorbic acid, vitamin E, lutein, β- carotene used either alone or in combination thereof.
3) A nutritional supplement as claimed in claim 1, wherein the methylating agent
is selected from the group comprising methionine, choline, folic acid, Vitamin
B6, Vitamin B12 used either alone or in combination thereof
4) A nutritional supplement as claimed in claim 1, comprising methionine, lycopene and green tea extract.
5) A nutritional supplement as claimed in claim 1, comprising methionine, choline, lycopene and green tea extract.
6) A nutritional supplement as claimed in claim 1, comprising methionine, choline, folic acid Vitamin B6, Vitamin B12, lycopene and green tea extract.
7) A nutritional supplement as claimed in claim 1, comprising green tea extract, vitamin E, ascorbic acid, selenium, lycopene, methionine, choline, folic acid Vitamin B6, Vitamin B12.
8) A nutritional supplement as claimed in claim 1, comprising green tea extract, vitamin E, ascorbic acid, selenium, lycopene, methionine, choline, folic acid Vitamin B6, Vitamin B12, lutein, β- carotene.
9) A nutritional supplement as claimed in claim 1, comprising green tea extract,
methionine, choline, folic acid, Vitamin B6, Vitamin B12.
10) A nutritional supplement as claimed in any of the preceding claims comprising lycopene between 0.5 and 25 mg.
11) A nutritional supplement as claimed in any of the preceding claims comprising green tea extract between 125 and 1000 mg.

12) A nutritional supplement as claimed in any of the preceding claims comprising methionine between 20 and 1000 mg.
13) A nutritional supplement as claimed in any of the preceding claims comprising choline between 2 and 50 mg.
14) A nutritional supplement as claimed in any of the preceding claims comprising folic acid between 0.2 and 1.5mg.
15) A nutritional supplement as claimed in any of the preceding claims comprising vitaminB6 between 0.5 and 50mg.
16) A nutritional supplement as claimed in any of the preceding claims comprising vitaminBn between 0.01 and 0.03mg.
17) A nutritional supplement as claimed in any of the preceding claims comprising ascorbic acid between 50 and 1000mg
18) A nutritional supplement as claimed in any of the preceding claims comprising Vitamin E between 5 and 250mg
19) A nutritional supplement as claimed in any of the preceding claims comprising selenium between 0.02 and 0.25mg.
20) A nutritional supplement as claimed in any of the preceding claims comprising lutein between 0.25 and 20mg and/or, β- carotene between 3 and 15 mg.
21) A nutritional supplement as claimed in any of the preceding claims which, comprises, folic acid between 0.2 to 1.5 mg, vitamin B6 between 0.5 to 50 mg, vitamin B12 between 0.01 to 0.03 mg methionine between 20 to 1000mg, choline between 2 to 50mg, lycopene between 0.5 to 25mg, ascorbic acid between 50 to lOOOmg, vitamin E between 5 to 250mg, selenium between 0.02 to 0.25 mg, green tea extract between 125 to 1000mg.
22) A nutritional supplement as claimed in any of the claims 1 to 21 in the form of tablets, hard gelatin capsules, soft gelatin capsules, rediuse, paste, pills, beads, dry syrup, sachet, liquid orals, powder, syrup, pharmaceutically acceptable oil, a solution or a suspension or similar conventional dosage forms.
23) A nutritional supplement as claimed in any of the claims 1 to 22 in the form of a tablet.

24) A process for the manufacture of a nutritional supplement as claimed in any of the claims 1 to 23 which process comprises incorporating green tea extract along with atleast one antioxidant and atleast one methylating agent in a pharmaceutically acceptable carrier to provide a nutritional supplement according to any of the claims 1 to 23.
25) A nutritional supplement as claimed in claim 1, comprising green tea extract along with atleast one antioxidants and atleast one methylating agents for retarding the progression of chronic diseases like hypertension, atherosclerotic cardiovascular diseases, cancer, diabetes, and neurological disorders and other disorders related to lack of nutrition.
Dated this 30th day of August, 2005
Dr. Gopakumar G. Nair Agent for the Applicant