FIELD OF INVENTION

The present invention relates to a synergistic composition comprising Luteolin and Myricetin for treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders. The present invention also relates to a method of employing the composition for treating diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders.

BACKGROUND OF THE INVENTION

Glucosidase enzymes are involved in several biological processes such as the intestinal digestion, the biosynthesis of glycoproteins and the lysosomal catabolism of the glycoconjugates (Naoki, A. et al. (1998) Homonojirimycin Isomers and N-Alkylated Homonojirimycins: Structural and Conformational Basis of Inhibition of Glycosidases, Journal of Medicinal Chemistry, 41 (14), p.2565-2571). Intestinal alpha-glucosidases are involved in the final step of the carbohydrate digestion to break down carbohydrates into simple sugars (monosaccharides) which can be readily absorbed from the intestine. As a result of the catalysis effect produced by alpha-glucosidase enzyme in the final step of digestive process of carbohydrates, its inhibitors can retard the uptake of dietary carbohydrates and suppress postprandial hyperglycemia, and could be therefore useful to treat diabetic and obese patients and also patients suffering from metabolic disorders (Sou, S. et al. (2000) Novel alpha-glucosidase inhibitors with a tetrachlorophthalimide skeleton, Bioorganic & Medicinal Chemistry Letters, 10 (10), p.1081-10840).
The alpha-glucosidase inhibitors are effective in lowering the insulin release, insulin requirement. Some inhibitors are also useful in lowering plasma lipids. Alpha-glucosidase inhibitors are increasingly finding therapeutic applications in metabolic disorders such as diabetes mellitus, obesity, hyperlipoproteinemia Type IV (Truscheit, E. et al. (2003) Chemistry and Biochemistry of Microbial a-Glucosidase Inhibitors, Angewandte Chemie International Edition in English, 20 (9), p.744-761) and cardiovascular diseases.

Historically, the knowledge gained from traditional medicinal practice and the screening of the extracts from plants and animals has yielded novel natural products which themselves act as potential bioactive agents and have the potential to be incorporated for the treatment of human diseases (Gullo, V. (1994) The Discovery of natural products with therapeutic potential. Boston: Butterworth-Heinemann; Cragg, G. et al. (1997) Natural Products in Drug Discovery and Development, Journal of Natural Products, 60 (1), p.52-60]. The screening of natural sources has led to the discovery of many clinically useful drugs that play a major role not only in the treatment of diseases such as diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, but also in the prevention of such diseases.

Therefore, increasing clinical importance of epidemics of diabetes and related diseases and cardiovascular diseases, has led to increased urgency to identify novel resources for active compounds and develop a large pool.

However, prior art literature reveals that not many studies have been conducted on natural compounds for inhibition of alpha glucosidase.

US7867989 describes the use of lichen metabolites for inhibition of alpha-glucosidase and treatment of diabetes, viral infections, fungal infections, autoimmune function disorders and obesity.

US 2004/0081711 describes a method for providing alpha-glucosidase inhibition to a subject by administration of a pharmaceutical composition comprising an alpha-glucosidase inhibitory agent selected from pipataline, sesamin, pellitorine, guineensine and brachystamide-B. The pharmaceutical composition is useful in treatment of diabetes mellitus, cancer, viral diseases such as hepatitis B and C, HIV and AIDS. The patent document further provides a process for the isolation of the alpha-glucosidase inhibitory agent from the plant source Piper longum.

US6299911 describes a Touchi extract with water and alcohol in the ratio of 1:5 to 5:1 acting as an alpha-glucosidase inhibitor. The extract reportedly inhibits alpha-glucosidase activity by at least 90.4%.

US2004/0171674 describes (-) - mesquitol and its analogs isolated from the medicinal plant Dichrostachys cinerea as alpha-glucoside inhibitory compounds. The invention further indentifies the use of the compounds and its analogs as broad based potential therapeutics as antihyperglycemic, antidiabetic, antiobesity, antiviral, anticancer, immunostimulants etc.

There have been many studies reporting the health benefits provided by bioflavonoids isolated from plant sources. The potential role of bioflavonoids in the prevention of cancers and cardiovascular disease and the treatment of inflammatory diseases has also been documented. Thousands of naturally occurring bioflavonoids derived from plant sources have been classified according to their chemical structure. The classes are flavones, isoflavones, flavan-3-ols and anthocyanidins. Flavones are divided into four groups: (1) flavones which includes, luteolin, apigenin and tangeritin; (2) flavonols, which includes, quercetin, kaempferol, myricetin, chrysin, rutin, rhoifolin, morin, fisetin, isorhamnetin, pachypodol and rhamnazin; (3) flavanones, which includes, galangin, hesperetin, naringenin, naringin, neohesperidin, hesperidin, narirutin, pruning, eriodictyol, homoeriodictyol; and (4) 3-hydroxyflavanones or 2,3-dihydroflavonols, which includes, dihydroquercetin and dihydrokaempferol.

Luteolin is the commonest bioflavonoids and is often found in the leaves of green pepper, tea, carrots, olive oil, peppermint, rosemary and navel oranges. Luteolin is a yellow crystalline compound. Based on preliminary research, it is thought to play a role in the human body possibly as an antioxidant, a free radical scavenger, a promoter of carbohydrate metabolism, an immune system modulator and anti-inflammatory agent.

Myricetin is a naturally occurring flavonol, and is found in many grapes, berries, fruits, vegetables, herbs, as well as other plants. Walnuts are a rich dietary source of Myricetin. Trace amounts of the bioflavonoid can also be found as glycosides. Myricetin is one of the phenolic compounds present in red wine. Myricetin also has antioxidant properties, and in vitro research suggests a role of high concentration of Myricetin in modification of LDL cholesterol.

WO 2003/026561 describes a composition comprising an aqueous extract of plants of the genus Brickellia for the treatment of diabetes mellitus. The composition consists of a flavonoid selected from the group consisting of luteolin, myricetin, dihydrokaemferol, apigenin, quercetin and mixtures thereof. However the document does not disclose the alpha-glucosidase inhibition activity of the composition.
WO 2008/140440 describes a stable flavonoid solution comprising (a) at least one flavonoid or flavonoid derivative and (b) a solvent system comprising (i) a mixture of alkyl glycols of two to seven carbon atoms, (ii) a polyol ether and (iii) at least one antioxidant. The flavonoid or flavonoid derivative is selected from the group consisting of luteolin, myricetin, baicalin, baicalein, silymarin and kushenol. The document further describes that the flavonoid solution is used for the treatment of a dermatologic condition. The document does not disclose alpha-glucosidase inhibition activity of the composition or use of the composition for treatment of metabolic disorders or other related diseases.

Thus, there persists a need for a composition containing natural products such as flavonids for treatment of diabetes or diabetes related disease, wherein the composition has alpha glucosidase inhibition activity.

SUMMARY OF THE INVENTION

An aspect of the present invention relates to a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and/or other metabolic disorders, wherein the composition comprises Luteolin and Myricetin, 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

The following drawings form part of the present specification and are included to further illustrate aspects of the present invention. The invention may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.

Figure 1 shows the performance of natural ingredients at 100 uM and 500 uM alpha glucosidase enzyme.
Figure 2 shows the performance of different concentrations (100 μM, 200 μM, 300 μM, 400μM and 500 μM) of Luteolin on alpha glucosidase.

Figure 3 shows the performance of different concentrations (100 μM, 200 μM, 300 μM, 400 μM and 500 μM) of Myricetin on alpha glucosidase.

Figure 4 shows the performance of synergy of Luteolin and Myricetin in different concentrations on alpha glucosidase.

DETAILED DESCRIPTION OF THE INVENTION

Those skilled in the art will be aware that the invention described herein is subject to variations and modifications other than those specifically described. It is to be understood that the invention described herein includes all such variations and modifications. The invention also includes all such steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of said steps or features.

Definitions
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 phrase "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human. Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the compound 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 solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions.

A "therapeutically effective amount" or "an effective amount", which are used interchangeably, is an amount sufficient to decrease or prevent the symptoms associated with the conditions disclosed herein, including diseases associated with diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders and related conditions contemplated for therapy with the compositions of the present invention.

"Alpha-glucosidase inhibitors" act to inhibit alpha-glucosidase, which is an enzyme that converts fructose to glucose, thus these inhibitors delay the digestion of carbohydrates. The undigested carbohydrates are subsequently broken down in the gut, thereby reducing the post-prandial glucose peak. Suitable examples include, but are not limited to, acarbose, voglibose and miglitol, emiglitate, MDL-25,637, camiglibose and MDL-73,945.

The concept of "combination therapy" is well exploited in current medical practice. Treatment of pathology by combining two or more agents that target the same pathogen or biochemical pathway sometimes results in greater efficacy and diminished side effects relative to the use of the therapeutically relevant dose of each agent alone. In some cases, the efficacy of the drug combination is additive (the efficacy of the combination is approximately equal to the sum of the effects of each drug alone), but in other cases the effect can be synergistic (the efficacy of the combination is greater than the sum of the effects of each drug given alone). As used herein, the term "combination therapy" means the two compounds can be delivered in a simultaneous manner, e.g. concurrently or wherein one of the compounds is administered first, followed by the second agent, i.e. sequential administartion. The desired result can either be subjective relief of one or more symptoms or an objectively identifiable improvement in the recipient of the dosage

As used herein the term "Diabetes" refers to high blood sugar or ketoacidosis, as well as chronic and general metabolic abnormalities arising from a prolonged high blood sugar status or a decrease in glucose tolerance. "Diabetes" encompasses both the Type 1 and Type 2 (Non Insulin Dependent Diabetes Mellitus or NIDDM) forms of the disease.

As used herein the terms "treatment of diabetes" and the "management of diabetes", are used interchangeably and do not necessarily mean a complete cure. It means that the symptoms or complications of the underlying disease are reduced, and/or that one or more of the underlying cellular, physiological, or biochemical causes or mechanisms causing the symptoms or complications are reduced. It is understood that "reduced", as used in this context, means relative to the untreated state of the disease, including the molecular state of the disease, not just the physiological state of the disease.

The term treatment of diabetes also includes within its scope the prophylactic treatment of an asymptomatic subject, such as a mammal, particularly a human, thought to be at risk of developing diabetes.

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. as a 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 "oral administration" includes oral, buccal, enteral and intra-gastric administration.

The term "Flavonoids" also called bioflavonoids and collectively known as Vitamin P or Citrin, are a class of plant secondary metabolites

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 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, Maturity Onset Diabetes of the Young (MODY), and gestational diabetes).

The present invention is not 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 also to be understood that the terminology used herein is for purposes of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only in the appended claims.

The present invention provides a novel synergistic combination of plant based ingredients which act synergistically at appropriate ratios for alpha glucosidase inhibition, used for treatment 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 Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts, thereof, wherein Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

The synergistic composition comprising Luteolin and Myricetin described herein may be compounded, for example with a pharmaceutically acceptable carrier for solid compositions such as tablets, pellets or capsules; capsules containing liquids; suppositories; solutions; emulsions; suspensions or any other form suitable for use. Suitable carriers include sterile water, sterile physiological saline, gum acacia, gelatin, starch paste, talc, keratin, colloidal silica, urea and the like. In addition, auxiliary, stabilizing, thickening, lubricating and coloring agents may be used.

Compositions for oral administration may be in the form of tablets, troches, lozenges, aqueous or oily suspensions, granules or powders, emulsions, capsules, syrups or elixirs. Orally administered compositions may contain one or more agents, such as sweetening agents which may be fructose, aspartame or saccharin; flavoring agents such as peppermint, oil of wintergreen or cherry, coloring agents and preserving agents to provide a pharmaceutically palatable preparation.

Further, compositions in tablet form may be coated to delay disintegration and absorption in the gastrointestinal tract thereby providing a sustained action over an extended period of time. Selectively permeable membranes surrounding an osmotically active driving compound are also suitable orally administered compositions. In these later platforms, fluid from the environment surrounding the capsule is imbibed by the driving compound, which swells to displace the agent or agent composition through an aperture.

Aqueous suspensions containing Luteolin and Myricetin or analogs of Luteolin and Myricetin described herein may also contain one or more preservatives, such as, ethyl or n-propyl-p-hydroxy-benzoate, one or more coloring agents, flavoring agents or sweetening agents.

The synergistic composition of the present invention described herein can be used in the form of a food additive, food supplement, dietary supplement in solid, semisolid or liquid form, which contains comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin.

The synergistic composition of the present invention may be incorporated into food product alone or in combination with another antidiabetic, antihyperglycemic (blood glucose lowering), or anti-lipidemic compound, in admixture with a carrier or an excipient suitable for oral administration.

Compositions for oral administration may be in the form of food product comprising the compositions of this invention.

Any conventional food processing technique may be used to achieve a product comprising an effective amount of the composition described herein. A lot of information on the art and technology of the various conventional food processing techniques and their practices in both the pet food and food industries is available and it is accordingly assumed that the general principles of these techniques are understood by a person skilled in the art.

The synergistic composition described herein used in the methods described herein is suited to control hyperglycemia in patients whose blood glucose cannot be controlled by diet alone.

The synergistic compositions of this invention may be administered by a variety of routes which may be oral, intramuscular, intravenous, subcutaneous, transdermal, rectal or even by inhalation. While the preferred mode of administration is oral, the precise mode of administration is left to the discretion of the practitioner. The compositions are advantageously effective when administered orally.

For the treatment of diabetes, hyperglycemia effecting a lowering of blood glucose, a synergistic composition comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or a pharmaceutically acceptable salt thereof described herein may be administered, together with another antidiabetic, antihyperglycemic or blood glucose lowering agent including, but not limited to insulin; a biguanide such as metformin or buformin; a sulfonylurea such as acetohexamide, chlorpropamide, tolazamide, tolbutamide, glyburide, glypizide or glyclazide; a PPAR gamma agonist including but not limited to the thiazolidinediones (such as troglitazone); an alpha-glucosidase inhibitor such as acarbose or miglitol; a Pradrenoceptor agonist such as PL-316, 243, cholestyramine, clofibrate, colestipol, fluvastatin, gemfibrozil, lovastatin, niacin, pravastatin, probucol, psyllium hydrophilic muccilloid, simvastatin and sodium dichloroacetate.

Although the present inventors do not wish to be limited to any particular mechanism of action to explain the hypoglycemic activity of the composition comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or a pharmaceutically acceptable salt thereof described herein, it is envisaged that they may advantageously be useful for treatment of both insulin-dependent or Type I diabetes and non-insulin-dependent or Type II diabetes.

It is a primary object of the present invention to provide a composition for treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders.

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

The present invention provides methods and compositions comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or a pharmaceutically acceptable salt thereof for modification and regulation of glucose metabolism, generally to reduce insulin resistance, hyperglycemia, hyperinsulinemia, obesity, overweight, cardiovascular disease and for the improvement of metabolism disorders, especially those associated with diabetes, and obesity.

The subject method of the present invention includes the administration to an animal, of a synergistic composition comprising two alpha glucosidase inhibitors in a specific ratio, which are able to inhibit the activity of alpha glucosidase thereby reducing breakdown of polysaccharides into monosaccharide.
The invention relates to two specific natural ingredients namely Luteolin and Myricetin. 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 Luteolin and Myricetin or their analogs in a ratio in the range of 1:99 to 99:1 preferably, 4:1 for the inhibition of alpha glucosidase enzyme.

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

The synergistic composition of the present invention having Luteolin and Myricetin in the weight ratio of 4:1 shows enhanced inhibition activity of alpha glucosidase.

Accordingly, the composition of the invention may be used alongside other therapies for the treatment of metabolic syndrome or Type II diabetes and its associated complications. The therapies include administration of biguanide drugs, for example metformin, phenformin and buformin, insulin, synthetic insulin analogues, amylin and oral antihyperglycemics. An example of an alpha-glucosidase inhibitor is acarbose or voglibose or miglitol.

The present invention demonstrates an enhanced alpha glucosidase inhibition using a synergistic composition comprising Luteolin and Myricetin in the weight ratio of 4:1. Surprisingly, it was found that the 4:1 weight ratio of Luteolin and Myricetin has a synergistic effect for 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/or other metabolic disorders, wherein the composition comprises Luteolin and Myricetin, 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein the weight ratio of Luteolin to Myricetin is 4:1.

In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the analogs are 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition comprises 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, 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 β-cells, metformin, and glucosidase inhibitors.
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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof in a weight ratio in the range of 4:1 to 1:4, wherein the composition further comprises insulin.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition optionally comprises Ml receptor antagonists.

In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition optionally comprises prolactin inhibitors.
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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs in a weight ratio between 4:1 and 1:4, wherein the composition optionally comprises agents acting on an ATP-dependent channel of P-cells.
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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition optionally comprises metformin.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition optionally comprises glucosidase inhibitors.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, 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.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of an immediate-release composition.
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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of a controlled-release composition.

In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of sustained-release orally-administrable compositions.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of topically-administrable compositions.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of liquid solutions.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of liquid sprays or throat sprays.

In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of lozenges.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of ointments or cream.

In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of foams or an aerosol.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of cough drops.
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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of dissolvable strips.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of a jelly or a lollipop.
In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of a mouthwash or a gargle.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of a lollipop.

In yet 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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of aqueous or oily suspensions.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of dispersible powders or an emulsion.

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 comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4, wherein the composition is in a form of a syrup or a lotion.
In another embodiment of the present invention, there is provided a process for the preparation of a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts, thereof, wherein the process comprises mixing Luteolin and Myricetin or analogs of Myricetin and Capsaicin in a solvent in a weight ratio between 4:1 and 1:4, and optionally adding a pharmaceutically acceptable carrier to obtain the composition.

In still another embodiment of the present invention, there is provided a process for the preparation of a synergistic composition for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders comprising Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts, thereof, wherein the process comprises mixing Luteolin and Myricetin or analogs of Myricetin and Capsaicin in a solvent in a weight ratio between 4:1 and 1:4, and optionally adding a pharmaceutically acceptable carrier to obtain the composition, wherein the solvent is DMSO.
An embodiment of the present invention provides a food, beverage or feed supplement comprising the synergistic composition of the present invention, wherein the synergistic composition comprises Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

In an embodiment of the present invention, there is provided a food comprising the synergistic composition of the present invention, wherein the synergistic composition comprises Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

In another embodiment of the present invention, there is provided a beverage comprising the synergistic composition of the present invention, wherein the synergistic composition comprises Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

In yet another embodiment of the present invention, there is provided a feed supplement comprising the synergistic composition of the present invention, wherein the synergistic composition comprises Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

Another embodiment of the present invention provides a food, beverage or feed product comprising the synergistic composition of the present invention, wherein the synergistic composition comprises Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

In still another embodiment of the present invention, there is provided a feed product comprising the synergistic composition of the present invention, wherein the synergistic composition comprises Luteolin and Myricetin or analogs of Luteolin and Myricetin and/or pharmaceutically acceptable salts thereof, wherein Luteolin and Myricetin or analogs of Luteolin and Myricetin are present in a weight ratio between 4:1 and 1:4.

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, photochemical, and combinations thereof.

Industrial Applicability
The present invention provides a useful synergistic composition for maintaining satisfactory blood glucose level control in patients suffering from diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders. The synergistic composition of the present invention for treating diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders comprising Luteolin and Myricetin 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 alpha glucosidase Inhibitors
The natural ingredients belonging to the group: Kaempferol, Luteolin, Myricetin, Quercetin, Caffeine and Capsaicin were selected for alpha glucosidase inhibition assay. The alpha glucosidase assay was carried out in a total reaction volume of 200 μL comprising assay buffer, substrate, inhibitor and enzyme. 0.1M phosphate buffer at pH 7 was used as the assay buffer as well as the solvent for the enzyme and substrate solutions. The assay components were added in their respective volumes and the reaction was started by adding the enzyme. The reaction was carried out at 37°C for one hour with the absorbance readings being measured every minute. The assay is based on the action of alpha-glucosidase enzyme on alpha-1,4 glucosidic bonds. The substrate used in the assay is para-nitrophenol tagged alpha-D-glucopyranoside. The enzyme acts on the substrate and releases p-nitrophenol, the absorbance of which is measured colorimetrically at 410nm. Enzyme preparation is carried out using rat intestinal acetone powder. 500 mg of rat intestinal acetone powder is weighed and 10 mL of ice cold assay buffer is added to it. The solution is vortexed gently and allowed to stand in ice for 2 minutes. The mix is then centrifuged at 8000 rpm for 8 minutes. The supernatant is transferred to a fresh tube and the pellet is discarded. The supernatant aliquots are used as the enzyme solution. The above procedure is carried out at 4°C and all the enzyme solutions are stored at -20°C. Voglibose (500 uM) is used as a positive control in the alpha glucosidase inhibition assay. Percentage inhibition was calculated as, % Inhibition = (Absorbance control - Absorbance Test)/ Absorbance control x 100.

The results are provided in Figure 1. Figure 1 shows the performance of natural ingredients at 100 uM and 500 uM alpha glucosidase enzyme. Figure 2 shows the performance of natural ingredients at 500 uM on DPP-IV. It was inferred from Figure 1 that out of all the natural ingredients used for the assay, Luteolin and Myricetin were able to inhibit nearly 50% alpha glucosidase enzyme at a concentration of 500 uM with respect to Voglibose (a known inhibitor of alpha glucosidase enzyme) which was taken as positive control.

Based on the results obtained, alpha glucosidase inhibition assay was performed using Luteolin and Myricetin at concentrations in the range of 100 μM to 500 μM. Figure 2 shows the performance of different concentrations (100 uM, 200 μM, 300μM, 400 μM and 500μM) of Luteolin on alpha glucosidase. Figure 3 shows the performance of different concentrations (100μM, 200 μM, 300μM, 400 μM and 500 μM) of Myricetin on alpha glucosidase.

It was inferred from Figure 2 that maximum % inhibition of alpha glucosidase enzyme was obtained with 500 uM concentration of Luteolin with respect to Voglibose (positive control). The % inhibition at 500 uM was calculated to be 65%.

It was inferred from Figure 3 that maximum % inhibition of alpha glucosidase enzyme was obtained with 500 uM concentration of Myricetin with respect to Voglibose (positive control). The % inhibition at 500 uM was calculated to be 50%.

Example 2
Synergy of Luteolin and Myricetin for enhanced alpha glucosidase inhibition Based on the results obtained, Luteolin and Myricetin were selected for synergy studies and alpha glucosidase inhibition assay was performed. The amount of Luteolin and Myricetin were taken as provided in Table 1.

Table 1: Amount of Luteolin and Myricetin taken for alpha glucosidase inhibition assay

The alpha glucosidase assay was performed as described in Example 1. The results are provided in Figure 4. Figure 4 shows the performance of synergy of Luteolin and Myricetin in different concentrations on alpha glucosidase. It was inferred from Figure 4 that maximum % inhibition of alpha glucosidase with respect to Voglibose was obtained in composition comprising 400 μM Luteolin and 100 μM Myricetin (weight ratio of 80:20). The % inhibition was calculated to be 83%.

Thus, it was demonstrated that Luteolin and Myricetin in a particular combination in the weight ratio of 80:20 show enhanced inhibitory activity of alpha glucosidase as compared to Luteolin and Myricetin alone. These results suggest that the synergistic effects of Luteolin and Myricetin in inhibiting alpha glucosidase activity can be used for development of pharmaceutical compositions for the treatment of diabetes, obesity, overweight, cardiovascular diseases and other metabolic disorders, and also 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 Luteolin and Myricetin, 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 Luteolin to Myricetin 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 comprising 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.