FIELD OF INVENTION

The present invention relates to a synergistic composition for treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders comprising Myricetin and Capsaicin. The present invention further relates a process for preparing the composition.

BACKGROUND OF THE INVENTION

Incretins are a group of gastrointestinal hormones that cause an increase in the amount of insulin released from the beta cells of the islets of Langerhans after eating, even before blood glucose levels become elevated. They also slow the rate of absorption of nutrients into the blood stream by reducing gastric emptying and may directly reduce food intake. They also inhibit glucagon release from the alpha cells of the Islets of Langerhans. Incretins are classified into two types: Glucagon-like peptide-1 (GLP-1) and Glucose-dependent insulinotropic polypeptide (GIP) (Drucker DJ and Nauck MA (November 2006). The incretin system: glucagon-like peptide-1 receptor agonists and dipeptidyl peptidase-4 inhibitors in type 2 diabetes. Lancet 368 (9548): 1696-705]).

Glucagon-like peptide-1 (GLP-1) is derived from the transcription product of the proglucagon gene. The major source of GLP-1 in the body is the intestinal L cell that secretes GLP-1 as a gut hormone. It is 30 amino acids in length. GLP-1 secretion by ileal L cells is dependent on the presence of nutrients in the lumen of the small intestine. The secretagogues (agents that cause or stimulate secretion) of this hormone include major nutrients like carbohydrate, protein and lipid. It is a potent anti-hyperglycemic hormone, inducing glucose-dependent stimulation of insulin secretion while suppressing glucagon secretion. GLP-1 appears to restore the glucose sensitivity of pancreatic (3-cells, with the mechanism possibly involving the increased expression of GLUT2 and glucokinase. GLP-1 is also known to inhibit pancreatic fJ-cell apoptosis and stimulate the proliferation and differentiation of insulin-secreting (3-cells. In addition, GLP-1 inhibits gastric secretion and motility. This delays and protracts carbohydrate absorption and contributes to a satiating effect.

Glucose-dependent insulinotropic peptide (GIP) is a 42-amino acid gastrointestinal regulatory peptide that stimulates insulin secretion from pancreatic beta cells in the presence of glucose. Glucose-dependent insulinotropic polypeptide (GIP) is secreted from the intestinal K-cells. However, the GIP receptor is widely distributed in peripheral organs, including the adipose tissue, gut, bone and brain, where GIP modulates energy intake, cell metabolism and proliferation, and lipid and glucose metabolism, eventually promoting lipid and glucose storage. In diabetes and obesity, the incretin effect of GIP is blunted, while the extrapancreatic tissues keep a normal sensitivity to this hormone.

Both GIP and GLP-1 are indispensable hormones to maintain normal glucose metabolism. However, patients with type 2 diabetes maintain normal or nearly normal GIP level in blood but show a reduced GLP-1 secretion level. GLP-1 is one of the strongest factors to increase insulin secretion and shows about 10 pmol/L of EC50 (half-maximal effective concentration).
Both GLP-1 and GIP are rapidly inactivated by the enzyme dipeptidyl peptidase-IV (DPP-IV). The synthesized GLP-1 is digested by dipeptidyl peptidase- IV (DPP- IV) and thereby, has 2 amino acids in the N-terminal excised and loses its activity. DPP- IV excises proline and alanine from the N-terminal of GLP- IV. However, simple change of their positions shows resistance against DPP- IV.

Classes of oral hypoglycemics that block DPP- IV are known as DPP- IV inhibitors. These DPP- IV inhibitors reduce glucagon and blood glucose levels. The mechanism of DPP- IV inhibitors is to increase incretin levels (GLP-1 and GIP) [Mcintosh C, Demuth H, Pospisilik J and Pederson, R. (2005). Dipeptidyl peptidase IV inhibitors: How do they work as new antidiabetic agents?. Regulatory Peptides 128 (2): 159-65; Behme Margaret T, Dupre John and McDonald Thomas J (2003). Glucagon-like peptide 1 improved glycemic control in type 1 diabetes. BMC Endocrine Disorders 3:3; Dupre J, Behme M T, Hramiak I M, McFarlane P, Williamson M P, Zabel P and McDonald T J. (1995).

Glucagon-like peptide I reduces postprandial glycemic excursions in IDDM. Diabetes 44 (6): 626-30], which inhibit glucagon release, which in turn increases insulin secretion, decreases gastric emptying, and decreases blood glucose levels.

WO 2006/074278 describes a composition that includes two compounds selected from a group of nine members, i.e., glucosidase inhibitor, an intestinal glucose transporter inhibitor, a glycation inhibitor, a nitric oxide production inhibitor, an aldose reductase inhibitor, a PPAR agonist, an adipocytokine activator, a glucose uptake enhancer, and a thermogenesis enhancer, and each compound is naturally occurring in a plant for treating diabetes or obesity. The document further discloses that the compounds are present in the form of a plant extract. However the document does not disclose the DPP-IV inhibition activity of the composition.

EP1925311 describes use of a plant extract or plant juice for the production of a pharmaceutical or nutritional preparation for treatment of metabolic syndrome, wherein the plant extract or juice is from thyme, oregano, glove, nutmeg, red clover, bay leaves or red onion. The metabolic syndrome includes diabetes. However the document does not disclose the DPP-IV inhibition activity of the pharmaceutical preparation.

Prior art literature reveals limited studies on DPP-IV inhibitory properties of natural compounds. Yogisha and Raveesha (2010) describe the DPP-IV inhibitory activity of methanolic extract of Mangifera indica leaves. The leaves of Mangifera indica were assessed for antidiabetic properties using normoglycaemic, glucose-induced hyperglycemia and streptozotocin (STZ) induced diabetic mice. The DPP-IV inhibitory activities of methanolic extract Mangifera indica was compared with Diprotin A, a reference standard (known DPP-IV inhibitor). However, the % inhibition of DPP-IV enzyme by methanolic extract of Mangifera indica leaves was 68% with respect to Diprotin A. The concentration of extract required to achieve 68% inhibition of DPP-IV was 320 ng/ml which is very high (S. Yogisha and K. A. Raveesha. Dipeptidyl Peptidase IV inhibitory activity of Mangifera indica, Journal of Natural Products, Vol. 3(2010):76-79).

Chakrabarti et al (Rituparna Chakrabarti, Singh Bhavtaran, Prakrith Narendra, Nimisha Varghese, Lalthanzama Vanchhawng, Mohamed Sham Shihabudeen H, Kavitha Thirumurgan; Journal of Natural Products, Vol. 4(2011): 158-163) describes the DPP-IV inhibitory activity of methanolic extract of the bark of tree turmeric Berberis aristata. The crude extract of the bark of Berberis aristata was assessed by in-vitro assay. The DPP-IV inhibitory activities of methanolic extract of the bark of Berberis aristata was compared with Diprotin A, a standard inhibitor of DPP-IV. However, the % inhibition of DPP-IV enzyme by methanolic extract of the bark of Berberis aristata was 65 % with respect to Diprotin A. This amount of % inhibition is quite low.
Thus, there is a need to provide a composition containing natural products for treatment of diabetes or diabetes related disease, wherein the composition has higher % of DPP-IV inhibition activity.

SUMMARY OF THE INVENTION

An aspect of the present invention relates to a synergistic composition for treatment of diabetes, obesity, overweight, cardiovascular diseases and/or other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

Figure 1 shows the performance of natural ingredients at 100 uM on DPP-IV.

Figure 2 shows the performance of natural ingredients at 500 uM on DPP-IV.

Figure 3 shows the performance of different concentrations (100 uM, 200 uM, 300 uM, 400 uM and 500 uM) of Myricetin on DPP-IV.

Figure 4 shows the performance of different concentrations (100 uM, 200 uM, 300 uM, 400 uM and 500 uM) of Capsaicin on DPP-IV.

Figure 5 shows the performance of synergy of Myricetin and Capsaicin in different concentrations on DPP-IV.

DETAILED DESCRIPTION OF THE INVENTION

Definition

For convenience, before further description of the present invention, certain terms employed in the specification, examples and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and understood as by a person of skill in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art.

The articles "a", "an" and "the" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article.

The term "plurality" means more than one.

The terms "at least two," "more than one" and "plurality" are used interchangeably.

The terms "comprise" and "comprising" are used in the inclusive, open sense, meaning that additional elements may be included. It is not intended to be construed as "consists of only".
Throughout this specification, unless the context requires otherwise the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated element or step or group of element or steps but not the exclusion of any other element or step or group of element or steps.

The term "including" is used to mean "including but not limited to". "Including" and "including but not limited to" are used interchangeably.

The term "treating" refers to the administration of an effective amount of a composition of the present invention to a subject, who has diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, or a symptom or a predisposition of such diseases, with the purpose to cure, alleviate, relieve, remedy, or ameliorate such diseases, the symptoms of them, or the predispositions towards them.

The term "administration" covers oral or parenteral delivery to a subject a composition of this invention in any suitable form, e.g., food product, beverage, tablet, and capsule.

The term "parenteral" refers to subcutaneous, intracutaneous, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection, as well as various infusion techniques.

The term "pharmaceutically acceptable carrier" is intended to refer to a carrier including but not limited to an excipient, diluent or auxiliary that can be administered to a subject as a component of a composition of the invention. Preferred carriers do not reduce the activity of the composition and are not toxic when administered in doses sufficient to deliver an effective amount of milk fat, a milk fat derivative or a component thereof, including, for example, cis-9, trans-11 CLA and TV A, or, when administered, of a lactoferrin polypeptide or functional variant or functional fragment thereof. The formulations can be administered orally, nasally or parenterally.

The term "oral administration" includes oral, buccal, enteral and intra-gastric administration.
The term "Flavonoids" also called bioflavonoids also collectively know as Vitamin P and citrin, are a class of plant secondary metabolites

The term "therapeutically effective amount" means any amount which, as compared to a corresponding subject who has not received such amount, results in improved treatment, healing, prevention, or amelioration of a disease, disorder, or side effect, or a decrease in the rate of advancement of a disease or disorder. The term also includes within its scope amounts effective to enhance normal physiological function.

The term "Synergistic effect/Synergize" refers to a combination of two or more treatments, which is more effective to produce advantageous results than the additive effects of these agents.
The term a "carrier" refers to, for example, a diluent, adjuvant, excipient, auxilliary agent or vehicle with which an active agent of the present specification is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water or aqueous saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions.
The term "effective amount" refers to the amount required to confer therapeutic effect. The interrelationship of dosages for animals and humans (based on milligrams per meter squared of body surface) is described by Freireich, et al. (1966). Body surface area can be approximately determined from height and weight of the subject. Effective doses also vary, as recognized by those skilled in the art, dependent on route of administration, excipient usage, and the like.
The term "a metabolic disorder" refers to any pathological condition resulting from an alteration in a patient's metabolism. Such disorders include those resulting from an alteration in glucose homeostasis resulting, for example, in hyperglycemia. According to this invention, an alteration in glucose levels is typically an increase in glucose levels by at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or even 100% relative to such levels in a healthy individual. Metabolic disorders include obesity and diabetes (e.g., diabetes type I, diabetes type II, MODY, and gestational diabetes).

The term "glucose transporter" refers to both glucose transporters and glucose co-transporters.
The term "glycation inhibitor" refers to non- enzymatic glycation inhibitors.

The present invention is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the invention, as described herein.
It is the primary object of the present invention to provide a composition for treatment of treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders.

It is an object of the present invention to provide a composition comprising Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof.
It is another object of the present invention to provide a composition comprising Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof, wherein Myricetin and Capsaicin or analogs of Myricetin and Capsaicin are present in a weight ratio between 4:1 and 1:4.

It is yet another object of the present invention to provide a composition comprising Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof, wherein Myricetin and Capsaicin or analogs of Myricetin and Capsaicin are present in a weight ratio of 3:2.

It is still another object of the present invention to provide a composition comprising Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof for treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders.

It is an object of the present invention to provide a food comprising a composition, wherein the composition comprises Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof.

It is another object of the present invention to provide a beverage comprising a composition, wherein the composition comprises Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof.

It is yet another object of the present invention to provide a feed supplement comprising a composition, wherein the composition comprises Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof.

It is still another object of the present invention to provide a feed product comprising a composition, wherein the composition comprises Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof.

The present invention provides a synergistic composition for the treatment of diabetes, obesity, cardiovascular diseases and other metabolic disorders comprising Myricetin and Capsaicin or their analogs.

Myricetin is a naturally occurring flavonol, a flavonoid found in many grapes, berries, fruits, vegetables, herbs, as well as other plants. Walnuts are a rich dietary source. Myricetin has antioxidant properties. In vitro research suggests that myricetin in high concentrations can modify LDL cholesterol such that uptake by white blood cells is increased.

Capsaicin is the active component of chili (Capsicum sps) and pepper (Piper nigrum), plants belonging to the genus Capsicum. Capsaicin and several related compounds are called capsaicinoids and are produced as a secondary metabolite by chili peppers. Pure capsaicin is a hydrophobic, colorless, odorless, and crystalline to waxy compound.

The compounds in the composition of this invention can be obtained by any suitable means. They can be obtained from commercial sources or, preferably, from various plants. Set forth below are a number of examples. Myricetin can be provided in an extract of onion, grape seed, berries, fruits, vegetables and herbs. Capsaicin can be provided in an extract of cayenne, black pepper or chilli.

Each of the extracts mentioned above can be prepared by first immersing a pulverized plant (or a part of a plant) in an aqueous solvent, an organic solvent, or a mixture of solvents. Examples of a suitable organic solvent include ethanol, dichloromethane, or hexane. The crude extract thus obtained can be filtered or centrifuged to remove any insoluble materials. A purified extract can then be obtained from the crude extract using liquid chromatography (e.g., high-pressure liquid chromatography) or other suitable methods. An extract can be produced either by a batch method or by a continuous method.

The composition of the present invention optionally comprises compounds selected from the group consisting of a-glucosidase inhibitor, an intestinal glucose transporter inhibitor, a glycation inhibitor, a nitric oxide production inhibitor, an aldose reductase inhibitor, a PPAR agonist, an adipocytokine activator, a glucose uptake enhancer, and a thermogenesis enhancer. The composition affects the following events that take place during carbohydrate metabolism in a human body: (1) hydrolysis of starch into monosaccharides, such as glucose; (2) entrance of glucose into the blood from small intestine; (3) reactions between glucose and proteins or nucleic acids in the blood; (4) binding of insulin to its receptors; (5) uptake of glucose into cells from the blood; and (6) oxidation of glucose in cells.

a-glucosidase inhibitor affects event (1). Specifically, it suppresses the activity of a-glucosidase, thereby reducing the hydrolysis of starch into oligosaccharides (lactose) or monosaccharides (e.g., glucose, fructose, or galactose). An intestinal glucose transporter inhibitor affects event (2). It reduces transport of monosaccharides from small intestine to the blood through glucose transporters located in cell membranes. Examples of glucose transporter include glucose transporter- 1 to glucose transporter- 12, proton myo-inositol transporter, and sodium-dependent glucose cotransporter-1 to sodium-dependent glucose cotransporter-6.

A glycation inhibitor, an aldose reductase inhibitor, and a nitric oxide production inhibitor affect event (3) by reducing the side effects generated by the high concentrations of glucose in the blood. Specifically, proteins and nucleic acids in the blood undergo glycosylation reactions in the absence of any enzyme. These reactions alter the structures and functions of the proteins and nucleic acids, thereby causing certain disorders. A high glucose concentration in the blood drives the glycosylation reactions. The glycation inhibitor suppresses non-enzymatic glycosylation reactions and therefore prevents certain diabetic disorders. An aldose reductase inhibitor prevents or reduces the action of aldose reductase. Aldose reductase is an enzyme present in the eye and many other parts of the body. It promotes conversion of glucose into sorbitol. Diabetic patients can have high concentrations of sorbitol in eye cells and nerve cells, which lead to retinopathy and neuropathy, respectively. An aldose reductase inhibitor reduces the formation of sorbitol, thereby preventing or delaying these complications of diabetes. A nitric oxide production inhibitor reduces excess production of inducible nitric oxide resulting from diabetes, which can cause diabetic complications and insulin resistance.

A PPAR agonist and an adipocytokine activator affect event (4) by enhancing the insulin action and ameliorating the insulin resistance. PPAR receptors belong to a family of nuclear receptors that regulate lipid metabolism. A PPAR agonist binds to PPAR receptors, thereby improving muscle insulin action and increasing insulin sensitivity. An adipocytokine enhancer improves adipocytokin secretion and up-regulates the adipocyte specific gene, thereby improving insulin sensitivity and glucose tolerance.

A glucose uptake enhancer affects event (5) by facilitating uptake of glucose from the blood to the cells, in which glucose is oxidized to generate energy. A thermogenesis activator affects event (6) by promoting oxidation metabolism of glucose (e.g., in TCA cycles) in the cells.

The composition of the present invention can be a dietary supplement or a pharmaceutical formulation. As a dietary supplement, additional nutrients, such as minerals or amino acids may be included. The composition can also be a beverage or a food product, e.g., tea, soft drink, juice, milk, coffee, cookie, cereal, chocolate, and snack bar. The composition of this invention can be in the form of a solution. For example, it can be an aqueous solution optionally containing a non-aqueous co-solvent, such as an alcohol. The composition can also be in the form of powder, paste, jelly, capsule, or tablet. Lactose and corn starch are commonly used as diluents for capsules and as carriers for tablets. Lubricating agents, such as magnesium stearate, are typically added to form tablets.

The composition of the present invention can be sweetened, if necessary, by adding a sweetener such as sorbitol, maltitol, hydrogenated glucose syrup and hydrogenated starch hydrolyzate, high fructose corn syrup, cane sugar, beet sugar, pectin, or sucralose.

The composition of the present invention comprises a pharmaceutically acceptable additives or a carrier. A pharmaceutically acceptable carrier includes wetting, emulsifying, or pH buffering agents or vehicles with which the pharmaceutical preparation can be contained in or administered. Examples are oils, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. Preferably, the agents of the pharmaceutical preparation are formulated as salt forms. Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric, butyric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

Preferably the composition of the present invention is an oral preparation, in particular preferred in form of a juice or tablet. The extracts, juices or compounds can be dried and formulated into tablets and administered orally. For example, the preparations can be formulated in the form of tablets, capsules, cachets, gelcaps, solutions, suspensions, and the like. Tablets or capsules can be prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulphate). The tablets may be coated by methods well-known in the art. Liquid preparations for oral administration may take the form of, but are not limited to, solutions, syrups or suspensions, or they may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, or fractionated vegetable oils); and preservatives (e.g., methyl or propyl- p-hydroxybenzoates or sorbic acid). The preparations may also contain buffer salts, flavoring, coloring, and sweetening agents as appropriate. Preparations for oral administration may be suitably formulated for slow release, controlled release, or sustained release of a prophylactic or therapeutic agent(s).

The composition, in any of the forms described above, is used for treating any type of diabetes or diabetes related diseases. Such disease include, but are not limited to type 1 diabetes, type 2 diabetes, hyperglycemia, type 1.5 diabetes, latent autoimmune diabetes (e.g., in adults), maturity onset diabetes, beta-cell apoptosis, hemochromatosis induced diabetes, impaired glucose tolerance, metabolic syndrome X, insulin resistance, cystic fibrosis related diabetes, polycystic ovarian syndrome, gestational diabetes, obesity, overweight, dyslipidemia, diabetic dyslipidemia, hyperlipidemia, hypertriglyceridemia, hyperlipoproteinemia, hypercholesterolemia, hypertension, essential hypertension, acute hypertensive emergency, arteriosclerosis, atherosclerosis, intermittent claudication (atherosclerosis oblilterens), cardiovascular disease, cardiomyopathy, cardiac hypertrophy, left ventricular hypertrophy, coronary artery disease, early coronary artery disease, heart insufficiency, exercise tolerance, chronic heart failure, mild chronic heart failure, arrhythmia, cardiac dysrythmia, syncopy, heart attack, myocardial infarction, Q-wave myocardial infarction, stroke, acute coronary syndrome, angina pectoris, unstable angina, cardiac bypass reocclusion, diastolic dysfunction, systolic dysfunction, non-Q-wave cardiac necrosis, catabolic changes after surgery, acute pancreatitis, irritable bowel syndrome, diabetic retinopathy, background retinopathy, preproliferative retinopathy, proliferative retinopathy, macular edema, cataracts, nephropathy, diabetic nephropathy, microalbuminuria, macroalbuminuria, neuropathy, diabetic neuropathy, distal symmetrical sensorimotor polyneuropathy, and diabetic autonomic neuropathy.

The subject method of the present invention includes the administration, to a subject, of a synergistic composition comprising two DPP-IV inhibitors in a specific ratio, which are able to inhibit the activity of DPP-IV thereby reducing the blood glucose level.

The invention relates to two specific natural ingredients namely Myricetin and Capsaicin or their analogs. Luteolin and Myricetin belong to class of flavonoids. The present invention provides a novel synergistic combination of two natural products of plant origin, such as Myricetin and Capsaicin or their analogs in a ratio in the range of 1:99 to 99:1 preferably, 3:2 for inhibition of DPP-IV enzyme.

The composition of the present invention uses a unique combination of natural ingredients which facilitate enhanced inhibition of DPP-IV. The present invention also provides an optimized combination of these natural ingredients at a desired ratio and molar concentration to inhibit DPP-IV enzyme for treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders.

The composition of the present invention having Myricetin and Capsaicin in the weight ratio of 3:2 shows enhanced inhibition activity of DPP-IV enzyme.

The present invention demonstrates an enhanced DPP-IV inhibition using a composition comprising Myricetin and Capsaicin in the weight ratio of 3:2. It is a surprise finding that at a given ratio, the effect is synergistic for its inhibition.

This work will have significance, especially to develop wellness and healthy indulgence food products for the management of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders.

The present invention provides a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders comprising Myricetin and Capsaicin or analogs of Myricetin and Capsaicin and/or pharmaceutically acceptable salts thereof, wherein Myricetin and Capsaicin or analogs of Myricetin and Capsaicin are present in a weight ratio between 4:1 and 1:4.

In an embodiment of the present invention, there is provided a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4

In another embodiment of the present invention there is provided a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio is 3:2.

The analogs of Myricetin and Capsaicin of the p[resent invention can be natural or synthetic analogs.

In an embodiment of the present invention, there is provided a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4, and a pharmaceutically acceptable additive or a carrier.

In still another embodiment of the present invention, there is provided a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4, wherein the composition comprises one or more other therapeutic agents selected from the group consisting of a-glucosidase inhibitor, insulin, Ml receptor antagonists, prolactin inhibitors, agents acting on an ATP-dependent channel of p-cells, metformin, an intestinal glucose transporter inhibitor, a glycation inhibitor, a nitric oxide production inhibitor, an aldose reductase inhibitor, a PPAR agonist, an adipocytokine activator, a glucose uptake enhancer, a dipeptidyl peptidase-IV inhibitor, and a thermogenesis enhancer.

In an embodiment of the present invention, there is provided a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4, and a pharmaceutically acceptable additive or a carrier, wherein the composition is in a form selected from the group consisting of an immediate-release composition, a controlled-release composition, sustained-release orally-administrable compositions, topically-administrable compositions, liquid solutions, liquid sprays, lozenges, throat sprays, ointments, solutions, foams, cough drops, dissolvable strips, a jelly, a mouthwash; a gargle, a lollipop, a gum, aqueous or oily suspensions, dispersible powders or granules, a syrup, an elixir, emulsions, a cream, a paste, a gel, a lotion, impregnated dressings, occularly-administrable compositions, inhalable particles, inhalable solutions, droplets, and aerosols.

The present invention further provides a process for the preparation of the composition, wherein the process comprising mixing Myricetin and Capsaicin or analogs of Myricetin and Capsaicin in DMSO in a weight ratio in the range between 4:1 and 1:4; and optionally adding a pharmaceutically acceptable carrier to obtain the composition.

An embodiment of the present invention provides a food, beverage or feed supplement comprising the synergistic composition of the present invention, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4.

Some embodiment of the present invention provides a method of treatment of diabetes, wherein the method comprises administering to a subject in need thereof a therapeutically effective amount of a synergistic composition comprising Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4, wherein the effective amount is sufficient to treat diabetes but not sufficient to suppress the immune system of the subject.

The method as claimed as disclosed in the present invention, wherein the composition is administered orally.

The composition of the present invention may be compounded with foods such as, but not limited to dairy products, grains, breads, meats, fruits, vegetables, rice and the like. The composition of the present invention can be compounded with additional ingredients. For example, the composition can be mixed with carbohydrates lipids, polypeptides, fatty acids, phytochemicals, and combinations thereof.

Carbohydrates that may be used in accordance with the teachings of the present invention include monosaccharides, disaccharides, oligosaccharides and polysaccharides such as, but not limited to trehalose, maltose, sucrose, dextrose, lactose, inulin, ribose, malt dextrin and the like. In one embodiment of the present invention the disaccharide is trehalose dihydrate, the oligosaccharide is fructo-oligosaccharide and the polysaccharide is malt dextrin.

Suitable lipids include, but are not limited to soy bean oil, olive oil, palm kernel oil, peanut oil, walnut oil, cannola oil and the like. Suitable polypeptides include whey protein, egg albumin, gelatin, milk proteins, and other animal and plant proteins. Finally, phytochemicals as used herein include such compounds as polyphenols, saponins, flavanoids, monoterpenes, allyl sulfides, lycopenes, carotenoids, polyactetylenes, silymarin, glycyrrhizin catechins and others.
The synergistic composition of the present invention can be taken orally as a bolus in the form of a gelatin capsule, pressed tablet, or gel cap. The composition can also be taken orally in the form of a liquid beverage. The liquid beverage may contain other ingredients such as, but not limited to flavor enhancers, sweeteners, viscosity enhancers and other food additives. The present invention can also be taken together with other foods either separately or compounded therewith.

Industrial Applicability

The present invention provides a useful composition for maintaining satisfactory blood glucose level control in patients suffering from diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders. The composition of the present invention for treating diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders comprising Myricetin and Capsaicin or their natural analogs as the main components is highly safe, useful and can be administered over a long term.

Further, the composition of the present invention can be used for development of anti¬diabetic and anti-obesity food, feed, and nutraceutical composition and for development of satiety enhancement foods.

Having thus described embodiments of the present invention it should be apparent to those skilled in the art that certain advantages have been achieved. It should also be appreciated that various modifications, adaptations, and alternative embodiments thereof may be made within the scope and spirit of the present invention.

EXAMPLES It should be understood that the following examples described herein are for illustrative purposes only and that various modifications or changes in light of the specification will be suggestive to person skilled in the art and are to be included within the spirit and purview of this application and the scope of the appended claims.

Example 1

Screening of Natural ingredients as DPP-IV Inhibitors

Some natural ingredients like Kaempferol, Luteolin, Myricetin, Quercetin, Caffeine and Capsaicin were selected for the DPP-IV inhibition assay. DPP-IV enzyme cleaves the chromogenic protein Glycyl-prolyl-p-nitroanilide [Gly-Pro-pNA] substrate at a position next to proline residue to give a yellow chromogen, p-nitroaniline. This results in an increase in absorbance at 405 nm. The natural ingredients (Kaempferol, Luteolin, Myricetin, Quercetin, Caffeine and Capsaicin) were tested individually for DPP-IV inhibition activity at concentrations in the range from l00uM to 500uM.

The DPP-IV inhibition assay was performed using chromogenic protein Glycyl-prolyl-p-nitroanilide [Gly-Pro-pNA] substrate and Human DPP-IV enzyme. The reaction mix consisted of 500uM of chromogenic protein Glycyl-prolyl-p-nitroanilide [Gly-Pro-pNA] substrate, lng of Human DPP-IV enzyme in a reaction volume of 200uL in 100 mM Tris-HC1 pH 8.0. The natural ingredients were taken at separate concentrations, lOOuM and 500uM. The natural ingredients were first pre-incubated with [Gly-Pro-pNA] substrate for 5 minutes and the reaction was initiated by addition of DPP-IV enzyme. The reaction mix was incubated at 37°C for 60 minutes, and absorbance was measured at 405 nm, which indicates the release of p-nitroaniline from Gly-L-Pro p-nitroanilide. Diprotin A (Ile-Pro-Ile) was used as a positive control in the DPP-IV inhibition assay at 25 uM. Percentage inhibition was calculated as, % Inhibition = (Absorbance Control - Absorbance Test)/ Absorbance Control x 100. The results are provided in Figure 1 and Figure 2. Figure 1 shows the performance of natural ingredients at 100 uM on DPP-IV. Figure 2 shows the performance of natural ingredients at 500 uM on DPP-IV. It was inferred from Figure 1 that none of the natural ingredients: kaempferol, Luteolin, myricetin, Quercetin, Caffeine and Capsaicin were able to inhibit DPP-IV enzyme at a concentration of 100 uM with respect to Diprotin A (a known inhibitor of DPP-IV enzyme) which was taken as positive control.

It was inferred from Figure 2 that Myricetin and Capsaicin were able to inhibit DPP-IV enzyme at a concentration of 500 uM '+. . m , mj mwith inhibition of 59% and 38% with respect to Diprotin A (a known inhibitor of DPP-IV enzyme) which was taken as positive control.

Based on the results obtained, DPP-IV inhibition assay was performed using Myricetin and Capsaicin at a concentration in the range from 100 uM to 500 uM. Figure 3 shows the performance of different concentrations (100 uM, 200 uM, 300 uM, 400 uM and 500 uM) of Myricetin on DPP-IV. Figure 4 shows the performance of different concentrations (100 uM, 200 uM, 300 uM, 400 uM and 500 uM) of Capsaicin on DPP-IV.

It was inferred from Figure 3 that maximum % inhibition of DPP-IV enzyme was obtained with 500 uM concentration of Myricetin with respect to Diprotin A which was taken as positive control. The % inhibition at 500 uM was calculated to be 60%.

It was inferred from Figure 4 that maximum % inhibition of DPP-IV enzyme was obtained with 500 uM concentration of Capsaicin with respect to Diprotin A which was taken as positive control. The % inhibition at 500 uM was calculated to be 36%.

Example 2

Synergy of Myricetin and Capsaicin for enhanced DPP-IV inhibition

Based on the results obtained, Myricetin and Capsaicin were selected for synergy studies. DPP-IV inhibition assay was performed using 500uM of chromogenic protein Glycyl-prolyl-p-nitroanilide [Gly-Pro-pNA] substrate, lng of Human DPP-IV enzyme in a reaction volume of 200uL in 100 niM Tris-HCl pH 8.0. The amount of Myricetin and Capsaicin were taken as provided in Table 1.

Table 1: Amount of Myricetin and Capsaicin taken for DPP-IV inhibition assay.

Myricetin and Capsaicin in the amount as provided in Table 1 were first pre-incubated with [Gly-Pro-pNA] substrate for 5 minutes and the reaction was initiated by addition of DPP-IV enzyme. The reaction mix was incubated at 37°C for 60 minutes, and absorbance was measured at 405 run, which indicates the release of p-nitroaniline from Gly-L-Pro p-nitroanilide. Diprotin A (Ile-Pro-Ile) was used as a positive control at 25 uM. Percentage inhibition was calculated as, % Inhibition = (Absorbance Control - Absorbance Test)/ Absorbance Control x 100.

The result is provided in Figure 5. Figure 5 shows the performance of synergy of Myricetin and Capsaicin in different concentrations on DPP-IV. It was inferred from Figure 5 that maximum % inhibition of DPP-IV with respect to Diprotin A was obtained in composition comprising 300 uM Myricetin and 200 uM Capsaicin. The % inhibition was calculated to be 83%.

Thus, it was demonstrated that Myricetin and Capsaicin in a particular combination in the weight ratio of 60:40 ratio shows enhanced inhibitory activity of DPP-IV as compared to Myricetin and Capsaicin alone. These results suggest that the synergistic effects of Myricetin and Capsaicin in inhibiting DPP-IV activity can be used for development of pharmaceutical compositions for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, for development of food and feed products for patients suffering from such disease and for development of satiety enhancement foods and nutraceutical composition.

I/We Claim:

1. A synergistic composition for treatment of diabetes, obesity, overweight, cardiovascular diseases and/or other metabolic disorders, wherein the composition comprises Myricetin and Capsaicin, analogs thereof, or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4.

2. The synergistic composition as claimed in claim 1, wherein the weight ratio of Myricetin to Capsaicin is 4:1.

3. The synergistic composition as claimed in claim 1, wherein the analogs are natural or synthetic analogs.

4. The synergistic composition as claimed in claim 1, wherein the composition optionally comprises pharmaceutically acceptable additives or a carrier.

5. The synergistic composition as claimed in claim 1, wherein the composition optionally comprises one or more other therapeutic agents selected from a group consisting of insulin, Ml receptor antagonists, prolactin inhibitors, agents acting on an ATP-dependent channel of P-cells, metformin and glucosidase inhibitors.

6. The synergistic composition as claimed in claim 1, wherein said composition is in a form selected from the group consisting of an immediate-release composition, a controlled-release composition, sustained-release orally-administrable compositions, topically-administrable compositions, liquid solutions, liquid sprays, lozenges, throat sprays, ointments, solutions, foams, cough drops, dissolvable strips, a jelly, a mouthwash; a gargle, a lollipop, a gum, aqueous or oily suspensions, dispersible powders or granules, a syrup, an elixir, emulsions, a cream, a paste, a gel, a lotion, impregnated dressings, occularly-administrable compositions, inhalable particles, inhalable solutions, droplets, and aerosols.

7. A food, beverage or feed supplement comprising the synergistic composition as claimed in claim 1.

8. A food, beverage or feed product comprising the synergistic composition as claimed in claim 1.

9. A method of treatment of diabetes, wherein the method comprises administering to a subject in need thereof a therapeutically effective amount of a synergistic composition as claimed in claim 1, wherein the effective amount is sufficient to treat diabetes but not sufficient to suppress the immune system of the subject.

10. The method as claimed in claim 9, wherein the composition is administered orally.