WO2008059310A1

WO2008059310A1 - Cinnamomum zeylanicum water extracts and their application in diabetes related conditions

Info

Publication number: WO2008059310A1
Application number: PCT/IB2006/003257
Authority: WO
Inventors: Morawala Patell Villoo
Original assignee: Avestha Gengraine Technologies Pvt. Ltd
Priority date: 2006-11-13
Filing date: 2006-11-13
Publication date: 2008-05-22

Abstract

The present invention relates to water based herbal extracts from Cinnamomun zeylanicum plant species with hypoglycemic activity, which is characterized in delaying of onset and / or management of diabetes by inhibiting the glucose absorption in the intestine and by mimicking and potentiating the insulin activity and also to a method for producing the extract and the use of the extract in the treatment of diabetes. The invention relates to the extraction process of derivation of the extract from Cinnamomun zeylanicum, which is characterized in that the solvent used is water. The herbal extracts comprise active ingredients that delay saccharide digestion or absorption thereby suppressing a rise in postprandial and / or fasting blood glucose levels.

Description

Cinnamomum zeylanicum water extracts and their application in diabetes related conditions

FIELD OF THE INVENTION:

The present invention relates to water based herbal extracts from Cinnamomun zeylanicum plant species with hypoglycemic activity, which is characterized in delaying of onset and / or management of diabetes by inhibiting the glucose absorption in the intestine and by mimicking and potentiating the insulin activity and also to a method for producing the extract and the use of the extract in the treatment of diabetes. The invention relates to the extraction process of derivation of the extract from Cinnamomun zeylanicum, which is characterized in that the solvent used is water.

The herbal extracts comprise active ingredients that delay saccharide digestion or absorption thereby suppressing a rise in postprandial and / or fasting blood glucose levels.

BACKGROUND OF THE INVENTION:

^'Diabetes mellitus is a common, serious disease characterized by hyperglycemia. The disease can be divided into two major subclasses: insulin-dependent diabetes mellitus (IDDM), also known as type I diabetes, and non-insulin-dependent diabetes mellitus (NIDDM), also known as type II diabetes (World Health Organization Study Group. Diabetes mellitus. WHO Tech. Rep. Ser. 727:1-113, 1985). IDDM results from insulin deficiency caused by cell-mediated autoimmune destruction of pancreatic beta cells, and generally develops in the young (Bach J F., Insulin-dependent diabetes mellitus as a beta cell targeted disease of immunoregulation. J. Autoimm. 8:439-463,1995). IDDM accounts for approximately 10-15% of the diabetic population worldwide (World Health Organization Study Group. Diabetes mellitus. WHO Tech. Rep. Ser. 727:1-113,1985). In contrast, NIDDM results from a variable combination of insulin resistance and insulin deficiency and generally develops in adults (Jun H S, et al, Pathogenesis of non-insulin- dependent (Type II) diabetes mellitus (NIDDM) Genetic predisposition and metabolic abnormalities. Advanced Drug Delivery Reviews 35:157-177, 1999; DeFronzo R A., The triumvirate beta cell, muscle, liver: a collusion responsible for NIDDM. Diabetes 37:667- 687, 1988). However, NIDDM can also develop at a younger age, as seen in the maturity- onset diabetes of the young (Pirart J., Diabetes mellitus and its degenerative complications: a prospective study of 4400 patients observed between 1947 and 1973. Diabetes Care 1:168-188, 1978). NIDDM accounts for over 85% of the diabetic population worldwide.

Factors responsible for causing diabetes are heredity and obesity. Heredity increases the susceptibility of beta cells to viral invasions or favor the development of autoimmune antibodies against the beta cells, thus leading to their destruction. Obesity decreases the number of insulin receptors in the insulin target cells throughout the body. Hence, the amount of insulin present is inadequate to induce its usual metabolic effects. In diabetic patients blood glucose levels goes as high as 1200 mg/dl are known to occur which is 12 times higher than the normal. Levels of 300 mg/dl to 500 mg/dl are common in diabetic patients. Thus the tendency of diabetes is to cause both extra-cellular and intracellular dehydration to develop (Guyton, Ca & Hall Ej (1996): In Text Book Of Medical Physiology, 9.sup.th Ed., Chapter 78, "Insulin, Glucagon And Diabetes Mellitus", (Prism Indian Edition), Prism-Saunders, Bangalore, Pp 980 -983). Diabetic Symptoms, which may arise due to pathological physiology of Insulin lack, are Polyuria, Polydipsia, Polyphagia, Asthenia and diminished utilization of carbohydrates for energy.

Complications of disease diabetes Mellitus involves many organs systems of the human body leading to many systemic complications like Diabetic Neuropathy, Diabetic Diarrhea, Urinary retention, Gustatory Swelling, Papillary Reflexes, Cardiac Autonomic Disturbances, Collagen Disturbances. Of these disorders Diabetic Neuropathy is the most common and affects patients at earlier stages. The usual methods for diagnosing diabetes are based on various chemical tests of urine and the blood viz, Urinary Sugar, Fasting Blood Glucose Level, Postprandial Blood Glucose Level (Glucose Tolerance Test) and Acetone breath. Reduction of carbohydrate absorption in the intestine of animals, especially humans, is nutritionally and medically of great importance. Reduction of absorption can for example facilitate body weight management, e.g. as part of a method of treating obesity, and can be advantageous for subjects suffering form diabetes or hypoglycemic state.

In Ayurveda several herbal ingredients are mentioned for the treatment of diabetes (Madhumeha). Herbal ingredients such Gurmar leaves {Gyrnnema sylvestre), Methi seeds {Trigonella foenumgrecum), Vijayasar heartwood (Pterocarpus marsupium), Jamun seeds (Eugenia jambolana), Karela (Momordica charantia) etc are few examples in this category.

The present invention relates to the extracts of Cinnamomun zeylanicum in the delaying the onset and / or management of diabetes and the related conditions thereof as the complications mentioned supra. The genus Cinnamomum is comprised of over 250 species of trees and shrubs from east and Southeast Asia to Australia (Bailey et al., 1976, Riffle 1998, Wagner et al. 1999). Cinnamomum spp. originates from the island Sri Lanka. It is also native to South-west India and the Tenasserim Hills of Burma. Related Cinnamomun species are found in Indonesia, Vietnam and China and are: Cinnamomum burmannii (Indonesian Cinnamomun), Cinnamomum cassia (Cassia), Cinnamomum loureirii (Vietnamese Cinnamomun), Cinnamomum zeylanicum (Ceylonese Cinnamomun).

Cinnamomun is a tropical evergreen tree growing up to 7m (56 ft) in its wild state. It has deeply-veined ovate leaves that are dark green on top, lighter green underneath. The bark is smooth and yellowish. Both the bark and leaves are aromatic. It has small yellowish- white flowers with a disagreeable odour that bear dark purple berries. It prefers a humid tropical climate at a low altitude. In cultivated plantations grow as small bushes, no taller than 3 m (10 ft), as the stems are continually cut back to produce new stems for bark. The outer bark, cork and the pithy inner lining are scraped off and the remaining bark is left to dry completely, when it curls and rolls into quills. Many of these are rolled together to produce a compact final product, which is then cut into uniform lengths and graded according to thickness, aroma and appearance.

Cinnamomun is one of the oldest herbal medicines known, having been mentioned in Chinese texts as long as 4,000 years ago. The first medicinal use of Cinnamomun was in Egypt and parts of Europe as far back as 500 BC. Cinnamomun is often used for medicinal purposes due to its unique properties. These main properties of Cinnamomun are astringent, warming stimulant, carminative, antiseptic, antifungal, anti-viral, blood purifier, and digestive aid. AU of these properties of Cinnamomun make it a good medicinal plant. Cinnamomun has many historical medicinal uses in different cultures. Some of these uses include treatment of diarrhea, arthritis, menstrual cramps, heavy menstruation, and yeast infections. Traditionally in many cultures Cinnamomun was taken as medicine for colds, flu, and digestive problems. Today Cinnamomun is used for many of these same traditional reasons. Often Cinnamomun is used as a nonessential addition to other remedies, than as a remedy by itself. Often this is because Cinnamomun is a stimulant to other herbs and the body, enabling herbal remedies to work faster. According to research, Cinnamomun is also known to be acting as an insulin substitute in type 2 diabetes. Cinnamomun contains bioactive components that scientists believed has the potential to either prevent or overcome diabetes.

A controlled trial comparing a placebo against Cinnamomun was recently reported in Diabetes Care 2003; 26:3215-8. In this trial doses of Cinnamomum cassia ranging from 1-6 grams a day provided in divided daily doses all produced some significant reductions in blood sugar levels, total cholesterol levels, triglyceride levels and finally even lower levels of LDL lipoproteins. But this study was limited to the Chinese Cinnamomun herb.

Cinnamomun has a positive effect on the glycemic control and the lipid profile in patients with diabetes mellitus type 2. The aim of this trial was to determine whether an aqueous Cinnamomun purified extract improves glycated hemoglobin AIc (HbAIc), fasting plasma glucose, total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL) and triacylglycerol concentrations in patients with type 2 diabetes. Man q et.al Eur J Clin Invest. 2006 May; 36(5):340-4. Recent studies conducted by Onderoglu S et al., in 1999 suggested that Cinnamomun bark and olive leaf has certain effects on streptozotocin-induced tissue injury. The effects on glycaemia were evaluated. Long-term administration of olive leaf caused significant improvement in tissue injury induced by streptozotocin treatment; the effect of Cinnamomun bark was less extent. No effects on blood glucose levels were detected. However, significant decreases in some increased biochemical and haematological parameters of streptozotocin-treated rats were observed. Aspartate aminotransferase, urea and cholesterol levels were significantly decreased by treatment with both plant materials, and alanine aminotransferase by treatment with olive leaf. Cinnamomun bark also caused a significant decrease in platelet counts. In addition, any visible toxicity, except decrease in body weight gain, attributable to the long-term use of plant materials was not established in normal rats. The data indicate that long-term use of olive leaf and Cinnamomun bark may provide benefit against diabetic conditions. Determination of underlying mechanism(s) of beneficial effects, toxicity to other systems and clinical assessments of related plant materials are major topics requiring further studies.

According to Imparl-Radosevich J et al. 2001 the bioactive compound(s) extracted from Cinnamomun potentiate insulin activity, as measured by glucose oxidation in the rat epididymal fat cell assay. Wortmannin, a potent PI 3'-kinase inhibitor, decreases the biological response to insulin and bioactive compound(s) from Cinnamomun similarly, indicating that Cinnamomun is affecting an element(s) upstream of PI 3 '-kinase. Enzyme studies done in vitro show that the bioactive compound(s) can stimulate autophosphorylation of a truncated form of the insulin receptor and can inhibit PTP-I₅ a rat homolog of a tyrosine phosphatase (PTP-IB) that inactivates the insulin receptor. No inhibition was found with alkaline phosphate or calcineurin suggesting that the active material is not a general phosphatase inhibitor. It is suggested, then, that a Cinnamomun compound(s), like insulin, affects protein phosphorylation-dephosphorylation reactions in the intact adipocyte. Bioactive Cinnamomun compounds may find further use in studies of insulin resistance in adult-onset diabetes. The effects of Cinnamomun extracts on blood glucose, triglyceride, total cholesterol, HDL cholesterol, and LDL cholesterol levels in people with type 2 diabetes by Khan et ah, in 2003. A total of 60 people with type 2 diabetes, 30 men and 30 women aged 52.2 +/- 6.32 years, were divided randomly into six groups.. Groups 1, 2, and 3 consumed 1, 3, or 6 g of Cinnamomun daily, respectively, and groups 4, 5, and 6 were given placebo capsules corresponding to the number of capsules consumed for the three levels of Cinnamomun. The Cinnamomun was consumed for 40 days followed by a 20-day washout period. After 40 days, all three levels of Cinnamomun reduced the mean fasting serum glucose (18-29%), triglyceride (23-30%), LDL cholesterol (7-27%), and total cholesterol (12-26%) levels; no significant changes were noted in the placebo groups. Changes in HDL cholesterol were not significant. The results of this study demonstrate that intake of 1, 3, or 6 g of Cinnamomun per day reduces serum glucose, triglyceride, LDL cholesterol, and total cholesterol in people with type 2 diabetes and suggest that the inclusion of Cinnamomun in the diet of people with type 2 diabetes will reduce risk factors associated with diabetes and cardiovascular diseases.

Further investigations done by Qin et α/., in 2004 determined whether Cinnamomun extract (CE) would improve the glucose utilization in normal male Wistar rats fed a high- fructose diet (HFD) for three weeks with or without CE added to the drinking water (300 mg/kg/day). In vivo glucose utilization was measured by the euglycemic clamp technique. Further analyses on the possible changes in insulin signaling occurring in skeletal muscle were performed afterwards by Western blotting. At 3 mU/kg/min insulin infusions, the decreased glucose infusion rate (GIR) in HFD-fed rats (60 % of controls, p < 0.01) was improved by CE administration to the same level of controls (normal chow diet) and the improving effect of CE on the GIR of HFD-fed rats was blocked by approximately 50 % by N-monometyl-L-arginine. The same tendency was found during the 30-mU/kg/min insulin infusions. There were no differences in skeletal muscle insulin receptor (IR)-beta, IR substrate (IRS)-I, or phosphatidylinositol (PI) 3 -kinase protein content in any groups. However, the muscular insulin-stimulated IR-beta and IRS-I tyrosine phosphorylation levels and IRS-I associated with PI 3 -kinase in HFD-fed rats were only 70 +/- 9 %, 76 +/- 5 %, and 72 +/- 6 % of controls (p < 0.05), respectively, and these decreases were significantly improved by CE treatment. These results suggest that early CE administration to HFD-fed rats would prevent the development of insulin resistance at least in part by enhancing insulin signaling and possibly via the NO pathway in skeletal muscle.

Taher M et al., in 2004 attempted to discover a small active compound that could promote adipogenesis, they investigated the ability of Cinnamomun (Cinnamomum zeylanicum) extracts to stimulate 3T3-L1 preadipocytes, In this study, they designed an experiment by replacing insulin with Cinnamomun extracts. The differentiated of 3T3-L1 adipocytes were monitored using oil red staining method. Induction of adipocyte formation by cinnamtannin Bl or water extract gave the similar effects to insulin activity in adipogenesis.

The subject matter of the current invention describes extracts isolated from Cinnamomum zeylanicum and other related species of Cinnamomum such as C.burmannii which was the result of the planned experiments conducted to test the hypoglycemic potential of the said extracts, further analysis of their potential positive hypoglycemic properties which include its potent inhibitory effect on glucose absorption in the intestine.

The present invention relates to methods and various compositions and / or formulations for treating and preventing diabetes and its associated symptoms and conditions. Specifically, the present invention relates to water based extracts obtained from the Cinnamomun plant sp based on extraction methods and nutraceutical formulations comprising the foregoing extracts for treating Type II Diabetes and related conditions. Further, the present invention provides methods for inhibiting the onset or reducing the onset potential of future or additional diabetic developments. The present invention is suited for treatment and prevention of diabetes as commonly experienced in mammals, and particularly humans.

Pharmaceuticals play a major role in the treatment of diabetes. Medications, such as Glycosidase inhibitors (Glucobai), Amylase, Glibenclamide, and Niacinamide are effective control drugs. Treatment of Type 1 diabetes functions to maintain blood glucose at near normal levels, and may require home blood glucose testing and multiple daily insulin injections. While the existing therapies have some efficacy, there remains a continuing need for alternative therapeutic preferably herbal remedies that can augment or replace the existing therapies.

For Type II diabetes patients, suggested treatments start with diet and exercise. If the two measures fail to bring blood sugar levels under control, anti-diabetic drugs may be prescribed. The most commonly prescribed drugs categories for treatment of Type II diabetes are 1) drugs that stimulate insulin secretion; and 2) drugs that sensitize cells to insulin action. Unfortunately, the side effects of these drugs can be unpleasant and distressing. Side effects include hypoglycemia, upset stomach, skin rash or itching, weight gain, kidney problems, fatigue, dizzy spells, gloating and diarrhea. Beyond the unpleasant side effects associated with traditional oral medication, there are two problems with such drugs. One is the sudden surge of insulin in response to the blockage of the potassium channels. Ideally, insulin should be secreted after a meal when the glucose level is high. However, insulin release stimulated by drugs may not happen at the right time, or may be excessive causing a great fall in blood sugar level, or hypoglycemia, a dangerous condition with neurological side effects. The other problem is that long-term drug use may cause degeneration of insulin granules causing them to have to work harder. Herbal medications would be naturally available with minimal or no side effects, lower costs and of equal or greater efficacy than the existing therapies and medications. Nevertheless, it would be preferable instead of treating diabetes with drugs. Natural means are generally much more acceptable to patients, which results in an increased compliance.

This background information is provided for the purpose of making known information believed by the applicant to be of possible relevance to the present invention. No admission is necessarily intended, nor should be construed, that any of the preceding information constitutes prior art against the present invention. DESCRIPTION OF FIGURES:

The present invention will become more fully understood from the detailed description given herein below, and the accompanying drawings which are given by way of illustration only, and thus are not limitative of the present invention, and wherein;

Figure 1: Shows the free radical scavenging ability of the Cinnamomum zeylanicum water based bark extract comparable to ascorbic acid standard.

Figure 2: Shows the percent polyphenol content of the Cinnamomum zeylanicum water based bark extract comparable to gallic acid standard.

Figure 3: Shows the IC50 (μg/ml) value for alpha-glucosidase (0.2 EU) inhibition of the Cinnamomum zeylanicum water based bark extract compared to acarbose standard in a reaction mixture containing 3mM Para-nitro-phenyl-D-glucose.

Figure 4: Shows Insulin mimetic activity of the 3.33 μg of Cinnamomum zeylanicum water based bark extract compared to that of 1 OnM Insulin using tritiated glucose as an energy source in 3T3 L-I adipocyte cell lines.

Figure 5: Shows Insulin sensitization activity of the 3.33μg of Cinnamomum zeylanicum water based bark extract compared to that of 1 OnM Insulin using tritiated glucose as an energy source in 3T3 L-I adipocyte cell lines.

Figure 6: Shows Insulin mimetic activity of the 3.33 μg of Cinnamomum zeylanicum water based bark extract compared to that of 1OnM Insulin using tritiated glucose as an energy source in C2C12 myocyte cell lines. Figure 7: Shows Insulin sensitization activity of the 3.33 μg Cinnamomum zeylanicum water based bark compared to that of 1 OnM Insulin using tritiated glucose as an energy source in C2C12 myocyte cell lines.

Figure 8: Shows the percent changes in postprandial blood glucose level of diabetic Sprague dawley rats at different levels of Cinnamomum zeylanicum water based bark extract intervention (50, 125 and 250 mpk) compared to that of baseline values over 5 weeks.

Figure 9: Shows the percent changes in fasting blood glucose level of diabetic Sprague dawley rats at different levels of Cinnamomum zeylanicum water based bark extract intervention (50, 125 and 250 mpk) compared to that of baseline values over 5 weeks.

SUMMARY OF THE INVENTION:

The primary objective of the present invention is to identify, test, characterize and screen extracts isolated from the plant species of family Lauraceae, preferably of the genus Cinnamomum for their potent inhibitory effects on diabetes.

Another object of the invention is to provide a method of treating diabetes comprising administering to a person in need thereof an anti- diabetic or hypoglycemic effective amount of a composition comprising the extracts of the plant Cinamniomum (Cinnamon).

A particular embodiment of the invention describes the method of a suitable aqueous or organic solvent-based extraction of a specific therapeutically important phytochemical extract.

The therapeutic potential of the said extracts has been studied and confirmed through standard in vitro cell free and cell based assays.

It is a more specific aspect of the invention to provide a novel method of treating, preventing diseases or conditions characterized by diabetes and related conditions thereof by administering a therapeutically effective amount of the composition containing the extract to a mammal in need of such therapy.

It is another object of the invention to provide a novel single medicinal extract or a combination of extracts thereof derived from Cinammomum spp is preferably orally administrable but the invention contemplates topical, intradermal, intramuscular, parenteral or intravenous administrations thereof.

In accordance with foregoing objects, and in accordance with the invention as embodied and broadly described herein, the present invention also features a nutraceutical formulation comprising the extracts isolated designed to treat diabetes and its associated or related conditions and symptoms, as well as to balance and normalize insulin output and subsequent glucose transfer and to help maintain these to be within proper ranges.

These and other objects of the invention will be more fully understood and apparent from the following description of the invention, the referenced drawings attached hereto and the claims appended hereto. Furthermore, the features and advantages of the invention may be learned by the practice of the invention or will be obvious from the description, as set forth hereinafter.

DETAILED DESCRIPTION OF THE INVENTION:

It will be readily understood that the components of the present invention, as generally described herein, could be arranged and designed in a wide variety of different configurations. Thus, the following more detailed description of the embodiments of the system and method of the present invention is not intended to limit the scope of the invention, as claimed, but is merely representative of the presently preferred embodiments of the invention.

Plant material suitable for preparation of the plant extract for inclusion of the therapeutic composition of the invention is derived from a potential plant administered to a person suffering from diabetes, which results in the lowering of the blood glucose level of the patient. Administration of the composition to the patient both prevented and treated incidences of clinical diabetes.

In accordance with a further embodiment of the present invention, the potential plant is a member of the family Lauraceae. In another embodiment of the invention, the potential plant is a member of the genus Cinammomum. It will be readily apparent to one skilled in art that other extracts capable of potential positive anti-diabetic properties could be isolated using similar techniques from a wide range of plants i.e., potential plants. The potential plants include all species of the family Lauraceae, including terrestrial, aquatic or other plants that can be subjected to standard extraction procedures such as those described herein in order to generate an extract that can be tested for its therapeutic abilities. The present invention is directed to a herbal medicinal composition comprising the foregoing plant extracts that can be administered to a personal suffering from diabetes which results in the lowering of the blood glucose level of the patient.

Cinnamomum is a tropical evergreen tree growing up to 7m (56 ft) in its wild state. It has deeply-veined ovate leaves that are dark green on top, lighter green underneath. The bark is smooth and yellowish. Both the bark and leaves are aromatic. It has small yellowish- white flowers with a disagreeable odour that bear dark purple berries. It prefers a humid tropical climate at a low altitude. In cultivated plantations grow as small bushes, no taller than 3. m (10 ft), as the stems are continually cut back to produce new stems for bark. The outer bark, cork and the pithy inner lining are scraped off and the remaining bark is left to dry completely, when it curls and rolls into quills. Many of these are rolled together to produce a compact final product, which is then cut into uniform lengths and graded according to thickness, aroma and appearance.

As used herein, "extract" refers to a concentrated preparation of the essential constituents of the medicinal plant. Typically, an extract is prepared by drying and powderizing the plant. Optionally, the plant, the dried plant or the powderized plant may be boiled in solution. The extract may be used in liquid form, or it may be mixed with other liquid or solid medicinal herbal extracts. Alternatively, the medicinal herbal extract may be obtained by further precipitating solid extracts from the liquid form. As used herein, "anti-diabetic" or "hypoglycemic" compound or composition generally refers to an agent that lowers blood glucose levels. If blood glucose level is decreased by at least about 100 mg/dl, then the compound is considered to be a hypoglycemic agent. The hypoglycemic or anti-diabetic effect can be measured by a variety of methods including, but not limited to, measuring the blood glucose levels, the rate of insulin binding to its receptor, the level of insulin secretion from pancreatic beta cells, and inhibition of glucohydrolase activity.

In one embodiment of the invention, there is provided a process for obtaining a plant extract possessing hypoglycemic properties, the process comprising (a) obtaining plant material from one or more plants (b) obtaining an extract from the plant material by contacting the plant material with an aqueous solvent, thereby providing one or more plant extracts (c) analyzing the plant extracts for free radical scavenging potential, Intestinal alpha-glucosidase inhibition potential, in-vitro screening of Cinnamomum zeylanicum water bark extracts for glucose uptake and the in-vivo efficacy studies.

Extraction of the plant material by solvent extraction process:

The plant material employed in the extraction process can be the entire potential plant, or it can be one or more distinct tissues from the plant for example, leaves, seeds, roots, stems, flowers, or various combinations thereof but preferably the seed of the plant. If desired the plant material can be treated prior to extraction, for example, by drying, freezing, lyophilizing, or some combination thereof. If desired, the plant material can be fragmented and/or homogenized by some means such that a greater surface area is presented to the solvent. For example, the plant material can be crushed or sliced mechanically, using a grinder or other device to fragment the plant parts into small pieces or particles, or the plant material can be frozen liquid nitrogen and then crushed or fragmented into smaller pieces.

The solvent used for the extraction process can be aqueous, alcoholic or organic, or a combination thereof. In one embodiment of the present invention, plant material is extracted with an aqueous solvent. Examples of suitable solvents include but are not limited to water, buffers, cell media, dilute acids or bases and the like. Examples of suitable alcoholic solvents include, but are not limited to methanol, ethanol, n-propanol, iso-propanol, 2-butanol, ter-butanol, and combinations thereof. But preferably water is used as solvent for extraction purposes. The coarse powder of herbal material is soaked in water for a fixed duration at room temperature with constant stirring in pharmaceutical grade solvent extractor. The extract is passed though a continuous centrifuge rotating at 3000rpm to separate extract supernatant and material debris. The clear supernatant obtained is concentrated to 1/10 volume using concentrator and is treated with food grade organic solvents in a definite proportion to remove any unpleasant smell, taste and color. The concentrated and organoleptically optimized extract is dried completely using either spray drier or vacuum drier to get powdered extract. Thus prepared dried extract is stored in airtight food grade plastic bins and the same is taken through several in-vitro cells free and cell based bioassay to validate the extract efficacy.

Various extraction processes are known in the art and can be employed in the methods of the present invention. The extract is generally produced by contacting the solid plant material with a solvent with adequate mixing and for a period of time sufficient to ensure adequate exposure of the solid plant material to the solvent such that inhibitory activity present in the plant material can be taken up by the solvent.

The solvent extraction process may be selected from direct and successive extraction types such as extraction from plant parts in soxhlet apparatus or in flasks at room temperature or at higher temperature with polar and/or non-polar solvent (s). Regardless of the number of extraction processes, each extraction process typically is conducted over a period of time between about 6 hours to 24 hours at room temperature. Adequate contact of the solvent with the plant material can be encouraged by shaking the suspension. The liquid fraction is then separated from the solid (insoluble) matter resulting in the generation of two fractions: a liquid fraction, which is the potential extract, and a solid fraction. Separation of the liquid and solid fractions can be achieved by one or more standard processes known to those skilled in art. The present invention contemplates the large-scale preparation of the selected plant extracts of the invention. Such extracts can be prepared on a commercial scale by repeating the extraction process that lead to the isolation of the extract of interest. The small-scale extraction procedure can simply be scaled up and additional steps of quality control can be included to ensure reproducible results for the resulting extracts.

Also contemplated by the present invention are the modifications to the small-scale procedure that may be required during the scale up for the industrial level production of the extract. Such modifications may include for example, alterations to the solvent being used or to the extraction procedure per se employed in order to compensate for variations that occur during the scale-up and render the overall procedure more amenable to industrial scale production, or more cost effective. Modifications of this type are standard in the industry and would be readily apparent to those skilled in the art.

In yet another embodiment of the subject invention, concentration of the purified extracts or partially purified extracts by solvent removal from the original extract and/or fractionated extract, and/or purified extract. The techniques of solvent removal are known to those skilled in the art and include, but are not limited to rotary evaporation, distillation (normal and reduced pressure), centrifugal vacuum evaporation (speed vac), and lyophilisation.

The potential extracts obtained thereof are concentrated and solubilised in an appropriate solvent preferably ethyl acetate. Examples of various other organic solvents include but are not limited to, di-ethyl ether, hexane, heptane, dichloromethane, ethyl acetate, butyl alcohol, ether, acetone and the combinations thereof.

Tests to determine the free radical scavenging potential of the Cinnamomum 70% bark extracts

Free radicals are constantly being generated in the body, as a result of the normal metabolic processes. Under physiological conditions, damage due to free radicals is countered by antioxidants. Sometimes, excessive free radical formation occurs in the body, and the antioxidant systems in the body cannot cope with the situations, i.e., the pro-oxidants overwhelm the antioxidants. This situation is known as oxidative stress. Thus, oxidative stress is a general term used to describe a state of potential oxidative damage caused by free radicals. Increasing evidence suggests that increased oxidative stress and changes in nitric oxide (NO) formation or activity play a major role in the complications of diabetes. The weight of experimental and human evidence supports a clear role for increased oxidant stress in many of the proposed biochemical pathways linked to the microvascular and macro vascular complications of diabetes. In particular, very recent evidence has underscored the particular role the elevated glucose plays in oxidative modification of LDL by a superoxide dependent pathway, and has demonstrated that, in people with poorly controlled IDDM, there is increased LDL oxidation associated with reduced antioxidant defenses.

There are several intriguing human studies, however, that show that administration of antioxidants stress plays a role in the reduced metabolic effects of insulin. In humans, the diabetogenic process appears to be caused by immune destruction of the beta cells; part of this process is apparently mediated by white cell production of active oxygen species. Diabetes can be produced in animals by the drugs alloxan and streptozotocin; the mechanism of action of these two drugs is different, but both result in the production of active oxygen species. Scavengers of oxygen radicals are effective in preventing diabetes in these animal models. Not only are oxygen radicals involved in the cause of diabetes, they also appear to play a role in some of the complications seen in long-term treatment of diabetes. Oberley et.al Free radicals and diabetes, Free Radic Biol Med. 1988; 5(2): 113-24. Free radical reactions and non-enzymatic glycosylation may play important roles not only in the development of diabetes but also in its complications. Hayakawa et al, Free radicals and diabetes mellitus, Nippon Ronen Igakkai Zasshi. 1990 Mar; 27(2): 149-54.

In the present invention the isolated extracts were used to estimate its free radical scavenging potency relative to ascorbic acid by using Calorimetric-DPPH method (Polterait O. (1997) Anti Oxidants and free-radical Scavengers of Natural origin Current Org. Chem. 1. 415-440). The Cinnamomim water based extract isolated from the barks of the plant showed nearly 15% of free radical scavenging potency equivalent to that of ascorbic acid.

Estimation of total polyphenols and its hypoglycemic effects.

Several studies show that apple polyphenols have a positive effect on diabetes and insulin resistance in animals and humans. In 1987, European researchers announced that an apple-derived polyphenol, phlorizin, "completely normalized insulin response" in diabetic rats. In 2004, scientists at the National Institutes of Health in the U.S. gave this same apple polyphenol to mice. Two weeks of treatment "significantly decreased blood glucose levels" in diabetic mice. Whole body fat mass was also "significantly reduced."

An Asian study in diabetic human volunteers showed that even weak polyphenols in apple juice produced "avoidance of sharp peaks" in blood glucose levels. Polyphenols from wine have favorable effects on glucose metabolism and insulin sensitivity, and many observational studies from around the world have shown that diabetes occurs less commonly among moderate drinkers than among abstainers. The polyphenolic polymers have anti-oxidant effects, which provides synergistic benefits to persons with various forms of diabetes.

In the current invention, the total polyphenol content of the Cinnamomun water based extract was estimated relative to gallic acid using Calorimetric - Singleton method (Singleton, VX. and Ropssi, J. A. Jr (1965). Calorimetry of total phenolics with phosphomolybdic - phosphotungstic acid reagent, Am. J. Enol. Vitig. 16: 144 - 158). The Cinnamomun water based bark extract showed 20 ± 2.25 % total phenol content equivalent to gallic acid clearly indicative of the potential beneficial effects the extracts possess with respect to the management of diabetes and its medicative properties.

Tests to confirm the Intestinal α-glucosidase inhibition potential

The inhibition of degradation of oligosacharrides (carbohydrates having 2 to 10 glucose residues connected by 1 -4 or 1 -6 α-D-glycosidic linkage) into monosaccharides by alpha- glucohydrolase-catalyzed enzymatic reactions was tested for Cinnamomum zeylanicum water based bark extract using Calorimetric - para-nitro-phenyl (pNP) release method using pNP-α-D-glucoside (Halvorson. H, 'Methods in enzymology' VoI 8, Academics Press, New York, 1966, p 559-562). The Cinnamomun water based bark extract showed greater α-glucosidase inhibition potential (IC50 value of 147.06 μg/ml) relative to the commercially available α- glucosidase inhibitor, acarbose (IC50 value of 146.55μg/ml) for 0.2 α-glucosidase enzyme units at standard enzymatic reaction conditions.

In-vitro screening of Cinnamomun water based bark extract for glucose uptake

Insulin-stimulated glucose uptake in adipose tissue and striated muscle is critical for reducing postprandial blood glucose concentration and the dysregulation of this process is one of the hallmarks of Type -II Diabetes mellitus (Non Insulin dependent). Oral therapies for Diabetes mellitus have emerged out of this interest and are widely used still today. But rather than acting by mimicking insulin, these drugs acts either by stimulating insulin release [Sulphonylurease], potentiating insulin action (thiazolidinedione) or lowering hepatic glucose production (biguanides). Various amounts of Cinnamomun water based extract (0.034μg to 33.4μg) are tested for insulin mimetic and sensitization effects with / without insulin. Radio labeled glucose is used to measure the changes in the level of glucose uptake activity of the adipocyte cells in response to treatment with samples in the presence or absence of insulin. The assay is performed in a 96-well microtiter plate format and the counts per minute are measured using a radioactive counter. The count per minute can be measured on a microtitre plate by radioactive counter. The results of the tests conducted are displayed in Figures 4-7.

Use of the extract as a therapeutic composition:

The present invention envisages the method of treating diabetes and other related diseases thereof by administering an effective amount of the therapeutic composition comprising the single plant extract or the screened plant extracts purified there from in combination. The therapeutic compositions of the invention can be administered alone or in combination with one or more standard anti-diabetic therapeutics. The present invention also contemplates the administration of sub-optimal doses of the therapeutic composition, for example, chemotherapeutic drug(s), in combination with the therapeutic composition. Thus,, in one embodiment of the present invention, in order to prepare a therapeutic combination, one or more plant extracts is first selected and then the efficacy of the extract(s) in controlling diabetes and maintaining glucose homeostasis is determined using standard techniques as one of those outlined above. The efficacy of the one or more plant extract alone is then compared to the efficacy of the one or more plant extract in combination with varying amounts of another component i.e., another plant extract. The invention also contemplates the combination the plant extract with another synthetic inhibitor. A combination that demonstrates therapeutic index in comparison to the individual properties is considered to be an effective combination.

For compositions comprising two or more plant extracts, various ratios of the constituent plant extracts are contemplated. By a way of example, for a composition comprising two plant extracts, for example, extract A and extract B, the ratio of extract A to extract B can vary anywhere between 1 :99 and 99:1. By "anywhere between 99:1 and 1:99" it is meant that the ratio of the two extracts can be defined by any ratio within this ratio can be between 98:2 and about 1:99 between about 98:2 and 2:98, between 97:3 and 1:99, between 97:3 and 2:98, between 97:3 and 3:97, etc. The present invention contemplates the ratio of the two extracts is between about 90:10 and 10:90, 80:20 and 20;80, 70:30 and 30:70, 60:40 and 40:60 or 50: 50.Analogous ratios are contemplated for compositions comprising more than two or more plant extracts.

The formulations of the present invention contain at least an effective amount of the therapeutic composition. The effective amount is considered to be that amount of the composition, in weight percent of the overall formulation, which must be present in order to produce the desired therapeutic effect. As would be apparent to one skilled in art, the effective amount may vary, depending upon, for example the disease to be treated and the form of administration. In general the therapeutic composition will be present in an amount ranging from about 1% to 100% by weight of the formulation, 10% to about 90% by weight of the formulation, 20% to about 80% by weight of the formulation, 30% to 70% by weight of the formulation, from about 40% to 60% by weight of the formulation and about 50% by weight of the formulation.

The present invention contemplates the use of the therapeutic compositions at various stages in the disease development and progression, including in the treatment of early stage, or advanced and/or aggressive stage of hyperglycemia, diabetes or related disorders. The administration of the therapeutic composition comprising the isolated and screened extracts to mammal having an early stage of the disorder can help to attenuate the progression of the disease.

The dosage of the therapeutic composition to be administered is not subject to defined limits, but .will usually be an effective amount. However it will be understood that the actual amount of the composition to be administered will be determined by a physician, in the light of the relevant circumstances, including the exact condition to be treated, the chosen route of administration, the actual composition administered, the age, the weight, and the response of the individual patient and the severity of the patient's symptoms. The dosage ranges are not intended to limit the scope of the invention in any way.

The therapeutic compositions comprising the plant extract are not limited to only those for humans but also include those for various animals, in particular, other mammals. Therefore, the food compositions include foods for animals such as cats, dogs, and the like pets, and the medical compositions include those for animals other than humans.

Modes of administration:

For administration to a mammal, the therapeutic composition can be formulated as a pharmaceutical or naturopathic formulation such as phytoceuticals or nutraceuticals, for oral, topical, rectal or parenteral administration or for administration by inhalation or spray. The phytoceutical or naturopathic formulation may comprise the one or more plant extracts in dosage unit formulations containing the conventional non-toxic physiologically acceptable carriers, adjuvants and vehicles.

The pharmaceutical or naturopathic formulations may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion hard or soft capsules, or syrups or elixirs. The therapeutic compositions of the invention may be formulated as phytoceuticals, or nutraceuticals. Phytoceuticals may optionally comprise other plant-derived components and can therefore be delivered by such non-limiting vehicles as teas, tonics, juices or syrups. Nutraceuticals contemplated by the present invention may provide nutritional and/or supplemental benefits and therefore be delivered, for example as foods, dietary supplements, extracts, beverages or the like. Phytoceutical and nutraceuticals can be administered in accordance with conventional treatment programs and/or may be a part of the dietary or supplemental program.

Formulations intended for oral use may be prepared according to methods known in art for the manufacture of pharmaceutical compositions and may contain one or more agents selected from the group of flavoring agents, coloring agents and preserving agents in order to provide palatable preparations.

Tablets contain the active ingredient in admixture with suitable non-toxic physiologically acceptable excipients including, for example, inert diluents, such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid, binding agents, such as starch, gelatine or acacia, and lubricating agents, such as magnesium stearate, stearic acid or talc. The tablets can be uncoated, or they may be coated by known techniques in order to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.

Various additives or carriers can be incorporated into the orally delivered pharmaceutical naturopathic formulations or the invention. Optional additives of the present composition include, without limitation, phospholipids, such as phosphatidyl glycerol, phosphotidyl inositol, phosphotidyl serine, phosphotidyl choline, phosphotidyl ethanolamine as well as phosphatide acids, ceramide, cerebrosides, sphingomyelins and cardiolipins.

Pharmaceutical or naturopathic formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatine capsules wherein the active ingredient is mixed with water or an oil based medium such as peanut oil, liquid paraffin or olive oil.

Oily suspensions may be formulated by suspending the plant extract(s) in a vegetable oil, for example, arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Flavoring agents may be added to provide palatable oral preparations. These formulations can be preserved by the addition of an anti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation suitable for an aqueous suspension by the addition of water provide the active ingredient in admixture with dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents, sweetening, flavoring and coloring agents may also be present.

In a further aspect of the invention there is provided a comestible, that is to say, a foodstuff comprising at least an extract of the invention, typically in dried form, such as in a lyophilised form. The skilled addressee will appreciate that such cosmetibles may contain more than one extract of the invention and may be used. Such foodstuffs may be used in a prophylactic manner and may contain further extracts having a similar function to the first added extract or further added extracts may be added that have a different prophylactic function. Thus a foodstuff could either comprise extracts that provide for a comestible having a single functional aspect, or a comestible may have a multi-functional prophylactic effect against two or more disease types. It is thought that a multi-functional role could be assigned to pharmaceutical formulations comprising two or more extracts possessing dissimilar therapeutic or prophylactic properties designed either for prophylaxis or for the treatment of more than one disease(s) in a mammal, particularly in a human.

The type of foodstuff or comestible to which at least an extract of the invention may be added includes any processed food such as confectionaries, baked products including breads such as loafs, and fiat breads such as pitta bread, naan bread and the like, cakes, snack foods such as museli bars, compressed dried fruit bars, biscuits, dairy products such as yoghurts, milk and milk-based products such as custards, cream, cheese, butter and creme fraiche, simulated dairy food product such as Elmlea products, fruits and vegetable juices, aerated drinks, such as carbonated soft drinks and non-aerated drinks such as squashes, soya milk, rice milk and coconut milk and the like, pastas, noodles, vegetables, seed and nut oils, fruited oils such as sunflower oil, rapeseed oil, olive oil, walnut, hazelnut, and sesame seed oil and the like, and frozen confectionaries such as ice cream, iced yoghurts and the like.

The present water based plant extracts can also be used for the management of other diabetic complications which are general or local diseases directly or indirectly caused by diabetes. Specific examples thereof are diabetic acidosis, diabetic xanthoma, diabetic myatrophy, diabetic ketosis, diabetic coma, diabetic stomach disorders, diabetic gangrene, diabetic ulcer, diabetic diarrhea, diabetic microangiopathy, diabetic uterosclerosis, diabetic cardiomyopathy, diabetic neuropathy, diabetic nephropathy, diabetic blister, diabetic cataract, diabetic dermatitis, diabetic scleredema, diabetic retinopathy, diabetic necrobiosis lipoidica, diabetic blood flow obstructions, etc

The invention will now be exemplified with reference to the following Examples section. It is to be understood that the examples are not to be construed as limiting the scope of the invention in any way.

The present invention relates to mixtures, which can be isolated from Cinnamomun bark (Lauraceae family) for the management of an important clinical problem like diabetes. The following examples set forth and present the effects of Cinnamomum species on both pre-existing diabetes conditions, as well as the preventative effects of Cinnamomum species against the onset of or contracting diabetes. These examples are not intended to be limiting in any way, but are merely illustrative of the beneficial, advantageous, and remedial effects of Cinnamomum species on diabetes. Other non-limiting examples of the present invention are described below.

Example I

Cinnamomum zeylanicum bark is selected and tested for quality by pharmacognosy techniques. The authentic plant material is procured for bioactive extraction. Good quality bark is macerated into solvent extractable seed powder of preferable size of lOOμm. The bioactive extract is prepared from the bark. Water is used as the solvent for the preparation of the bioactive extract. The extraction process is carried out in a solvent extraction vessel of 100-liter capacity. The extraction process is undertaken at a temperature of 25⁰C and for duration of 240 minutes. The extraction is carried out with constant stirring at the rate of 200 rpm allowing the heavier particles in the extraction mixture to settle down and later the mixture is allowed to stand undisturbed for 120 minutes. Later the liquid phase of the extract is clarified by centrifugation. The centrifugation process is undertaken at 3000 rpm and at room temperature. The clarified extract was then concentrated to 1/10 of the total volume. The concentrated extract (two parts) is mixed well with a moderately non-polar solvent such as ethyl acetate (one part) in an extraction vessel for 60 minutes and left undisturbed for 120 minutes. The lower water phase is then separated and dried completely to powder using either spray drier or vacuum dried under reduced pressure. The powdered extract is then stored in an airtight plastic container. This extract is further used for all the biochemical, physiochemical, heavy metal, natural toxin and detailed efficacy study.

Example 2: Free radical scavenging potential

Cinnamomun water based bark extract isolated by the above mentioned method is tested to estimate its free radical scavenging potency relative to ascorbic acid by using Calorimetric-DPPH method (Polterait O. (1997) Anti Oxidants and free-radical Scavengers of Natural origin Current Org. Chem. 1. 415-440). The Cinnamomun water based bark extract showed 15 ± 2.5 % free radical scavenging potency equivalent to ascorbic acid upon comparison, which is depicted in figure 1.

Example 3: Total polyphenols

The total polyphenol content of the Cinnamomun water based bark extract is estimated relative to gallic acid using Calorimetric - Singleton method (Singleton, V.L. and Ropssi, J.A. Jr (1965). Calorimetry of total phenolics with phosphomolybdic - phosphotungstic acid reagent, Am. J. Enol. Vitig. 16: 144 - 158). The Cinnamomun water based bark extract showed 21 + 2.25 % total phenol content equivalent to gallic acid, which is shown in figure 2.

Example 4; Intestinal α-glucosidase inhibition potential

The inhibition of degradation of oligosacharrides (carbohydrates having 2 to 10 glucose residues connected by 1-4 or 1-6 alpha-D-glycosidic linkage) into monosaccharides by alpha-glucohydrolase-catalyzed enzymatic reactions is tested for Cinnamomun water based bark extract using Calorimetric - para-nitro-phenyl (pNP) release method using pNP-a-D-glucoside (Halvorson. H, 'Methods in enzymology' VoI 8, Academics Press, New York, 1966, p 559-562). The Cinnamomum water based bark extract showed appreciable α-glucosidase inhibition potential (IC50 value of 68.97μg/ml) relative to the commercially available - α glucosidase inhibitor, acarbose (IC50 value of 146.55μg/ml) for 0.2 α-glucosidase enzyme units at standard enzymatic reaction conditions. The results are depicted in figure 3.

Example 5: In-vitro screening of Cinnamomun water based bark extract for glucose uptake

Insulin-stimulated glucose uptake in adipose tissue and striated muscle is critical for reducing postprandial blood glucose concentration and the dysregulation of this process is one of the hallmarks of Type -II Diabetes mellitus (Non Insulin dependent). Oral therapies for Diabetes mellitus have emerged out of this interest and are widely used still today. But rather than acting by mimicking insulin, these drugs acts either by stimulating insulin release [Sulphonylurease], potentiating insulin action (thiazolidinedione) or lowering hepatic glucose production (biguanides). Various amounts of Cinnamomun water based extract (0.034μg to 33.4μg) are tested for insulin mimetic and sensitization effects with / without insulin. Radio labeled glucose is used to measure the changes in the level of glucose uptake activity of the adipocyte cells in response to treatment with samples in the presence or absence of insulin. The assay is performed in a 96-well microtiter plate format and the counts per minute are measured using a radioactive counter. The count per minute can be measured on a microtitre plate by radioactive counter.

The amount that showed best insulin mimetic and sensitization activity in 3T3L-1 adipocyte cells and C2C12 myocyte cells for glucose uptake was around 0.334μg. The method followed for screening is as follows:

Preadipocytes (3T3L-1) and premyocytes (C2C12) are cultured in DMEM containing 10%FCS, 4mM Glutamine, 2 % NaHCO3 and antimycotic, in an atmosphere of 5% CO2 at 370C, separately. Myoblasts are cultured up to 80% confluency and the cells are sub- cultured at three-day intervals. 20,000 of preadipocytes and myocytes are seeded separately in each well of a 96 well plate and differentiated for 48 hours in DMEM:F12(1 :1), 0.5mMIBMX, 0.25mM Dexmethasone and lug Insulin for 48hrs followed by incubation with lug of Insulin for δhours. The ability of the plant extract to induce glucose uptake is tested in two different ways 1) glucose uptake in presence of insulin (extract + insulin) and 2) glucose uptake in absence of insulin (extract alone). Therefore incubate in duplicate (one set to evaluate glucose uptake in presence of insulin i.e. extract + insulin and other set without insulin i.e. extract alone) with different concentration of extracts (300μg/well, 30μg/well, 3μg/well and 0.3μg/well) in triplicates for 18 hours at 370 C and 5%CO2 lOOμl of DMEM. The medium is then removed and the cells are incubated with KRH buffer (100 microliters) at 37⁰C and 5% CO2 for 10 minutes. Cells are incubated with insulin. For standard insulin response incubate cells with 5nM, 10 nM, 25 nM, 50 nM and 10OnM in KRH buffer in triplicate. To one set of wells to be treated with extract +insulin, 5nM of insulin in KRH buffer is added and in other set of well treated with extract alone, lOOμl of KRH buffer is added for 15 minutes at 37° C and 5%CO₂. Glucose uptake reaction is initiated by adding 0.1 rnM 2-deoxy glucose containing 2-deoxy [3H] glucose (final concentration 12.2 kBq/ml) and incubated for 1 hour at 37⁰C and 5%CO₂. assay is terminated by adding 40 μM Cytochalasin B. The cells are washed three times with ice-cold KRH buffer (lOOμl). KRH buffer is removed and 20 μl 1% Triton X is added to each well to lyse the cells and incubate for 10 min at 37°C and 5%CO₂. 200μl of Aqualite is added per well and the supernatant is transferred back to the plates and counted on a micro-titer plate radioactive counter. The results obtained for insulin mimetic and sensitization potential of Cinnamomun water based bark extract is depicted in figure 4 to figure 7 using differentiated adipocyte and myocytes. All the observed values of glucose uptake activity are blank corrected using the control (cells alone background value). These values are normalized with MTT cell viability assay values for the corresponding extracts. The degree of insulin mimetic/sensitization activity of each sample concentration is calculated as a percentage of that observed using 1OnM insulin alone.

Example 6; In-vivo efficacy screening using Sprague dawley rats

In-vivo efficacy screening of Cinnamomun water based bark extracts was done using Sprague dawley rats and the procedure undertaken is as follows:

Sprague dawley rats weighing ~250g with a variation of + 20% of the mean weight are selected for in-vivo efficacy screening of Cinnamomun water based bark extracts. The identification is undertaken by the cage tag and the corresponding picric acid color body markings. The number of animals selected in a group is five and kept in a experimental room after veterinary examination. The route of administration is oral gavage. The rats are fed with 5 % glucose water for two days before STZ injection to avoid death due to hypoglycemic shock. Rats are fasted for 12 hours and injected intraperitoneally with 45mg/kg-body weight of streptozotocin. Blood collection is undertaken with CO2 anesthesia and FBG analysis before STZ injection is performed which serves as the normal baseline (60-120mg/dl) for the animals. Citrate buffer (Ph of 4.5) is used as the vehicle. STZ is a photo, temperature and Ph sensitive chemical; hence the animals are injected with in 45 minutes of the dose formulation. Animals initially become hypoglycemic for the first 3 days because of the insulin surge into the blood stream. Gradually from day 4 animals attain hyperglycemia. On the 6^th day blood collected by retro orbital (ROP) plexus method in heparin as the anti-coagulant, under CO2 anesthesia and analyzed for postprandial glucose levels. Animals are fasted for 12 hours on Day 7 after STZ injection, Day 8 blood collected in heparin as the anti-coagulant, under CO2 anesthesia and analyzed for FBG. Animals that have the range of FBG beyond 250 mg/dl are considered for the study.

Example 7. Screening of Cinnamomun water based bark extract at 50, 125 and 200 mg/kg-body weight.

Selection, randomization and grouping of the animals into different treatment groups with the FBG range of 250 mg/dl is undertaken as above. The dosing formulations are prepared freshly each day 0.5% CMC was used as the vehicle. The test article Cinnamomun water based bark extract, is administered by oral gavage to each rat daily, for 35 consecutive days. The animals were dosed at approximately the same time each day where possible using a stainless steel intubation needle fitted onto a suitably graduated glass syringe. The dosage volume administered to individual rat was adjusted according to its most recently recorded body weights. Treatment in this manner continued once a day, seven days a week, for a total period of 35 days. Vehicle control group animals are treated with the vehicle only at the same dosage volume i.e. 10 ml/kg body weight. The Groups include: Gl-Vehicle control, G2-Diabetic control, G-3 Diabetic animals treated with Pioglitazone (Standard anti-diabetic drug) 20mg/kg-body weight. G- 4 Cinnamomun water based bark extract 50 mg/kg-body weight, G-5 Cinnamomun water based bark extract 125 mg/kg-body weight, G-6 Cinnamomun water based bark extract 200 mg/kg-body weight. Throughout the study, all cages were checked early on each working day and again in the afternoon and evening to look for dead or moribund animals to allow necropsy examination to be carried out during the working hours of that day. All rats that will be killed in extremes, or found dead in the cage were subjected to detailed necropsy examination. All signs of ill health, together with any behavioral changes or reaction to treatment are recorded for individual animals. Dated and signed records of appearance, change and disappearance of clinical signs are maintained in clinical history sheets for individual animals. The parameters that are observed or included in the study were postprandial glucose, fasting blood glucose (FBG), body weight and feed consumption. Blood collection by ROP on week 1 ,2 & 5 for the above said parameters. The animals selected for the study are treated with the extract at 3 different dose levels for 35 days. Necropsy Examination: On completion of 35 days of treatment period, all surviving rats will be sacrificed by exsanguinations under CO2 anesthesia. Complete necropsies were carried out on all animals including those, which died during the study.

The postprandial blood glucose lowering potential of Cinnamomun water based bark extract extracts at different concentrations are illustrated in figure 8. The diabetic control rat group showed 240% increase in their postprandial blood glucose levels compared to the baseline values of the same group at the start of the study. While the groups treated with pioglitazone, Cinnamomum zeylanicum water based bark extract at 50, 125 and 200mpk (milligram per kg body weight) has shown significant percent change from their baseline values. The results have been represented in Table No. 1

Table 1: Postprandial Blood Glucose levels

The fasting blood glucose lowering potential of the Cinnamomum water based bark extracts at different concentrations are illustrated in figure 9. The diabetic control rat group showed 246% increase in their fasting blood glucose levels compared to the baseline values of the same group at the start of the study. The results have been represented in Table No. 2

Week 1 (% change from Week 2 (% change from Week 5 (% change from

Groups Base line) Base line) Base line).

12.43 12.25 9.42

G1

87.78 115.15 246.60

G2

-30.99 -44.57 31.37

G3

-5.06 -47.85 0.69

G4

9.46 -56.62 14.60

G5

15.29 -9.24 5.78

G6

Table 2: Fasting Blood Glucose Levels

While the invention has been described in connection with specific and preferred, embodiments thereof, it is capable of further modifications without departing from the spirit and scope of the invention. This application is intended to cover all variations, uses, or adaptations of the invention, following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains, or as are obvious to persons skilled in the art, at the time the departure is made. It should be appreciated that the scope of this invention is not limited to the detailed description of the invention hereinabove, which is intended merely to be illustrative, but rather comprehends the subject matter defined by the claims. Although the present invention has been described in considerable detail with reference to certain preferred versions thereof, other versions are possible. Therefore, the spirit and scope of the appended claims should not be limited to the description of the preferred versions contained herein.

Claims

Claims:

1. A method of treating or preventing diabetes by administering to a mammal a therapeutically effective non-toxic amount of a water based extract derived from the plant Cinnamomum characterized of hypoglycemic activity.

2. A .method of obtaining a plant extract capable of delaying the onset and / or management of diabetes and related conditions thereof comprising the following steps: a. Obtaining plant material from one or more parts of the plant Cinnamomum preferably the bark. b. Obtaining an extract from the plant material by contacting the plant material with an aqueous solvent preferably water thereby providing one or more plant extracts possessing hypoglycemic activity. c. Analyzing the plant extracts for free radical scavenging potential, total polyphenols content, α-glucosidase inhibition potential and for insulin mimetic and sensitization activity. d. Selecting plant extracts having one or more of these activities.

3. A method according to claim 1 and 2, wherein the plant extracts are derived from the plant Cinnamomum.

4. A method according to claim any of the preceding claims wherein, the plant extracts are extracted from leaves, seeds, roots, stems, flowers, or various combinations thereof but preferably the bark of the plant.

5. A method according to claim 1 and 2, wherein the extract possessing hypoglycemic activity is derived by contacting the plant material with a solvent for a time period sufficient to ensure adequate exposure of the solid plant material to the solvent such that the potential extract is taken up by the solvent.

6. A method according to the preceding claim, wherein the solvent used for the extraction process is water.

7. A method according to claim 1 and 2, wherein the bioactive extract comprises at least 15 % free radical scavenging potential equivalent to ascorbic acid.

41

8. A method according to claim 1 and 2, wherein the bioactive extract comprises at least 22% of total polyphenols content equivalent to that of gallic acid.

9. The successful plant extracts derived from the extraction process demonstrating a) insulin mimetic activity b) insulin sensitizing activity c) alpha glucosidase activity in combination with a pharmaceutically acceptable carrier.

10. A method according to any one of the claims 1-9, wherein the said extract in a food stuff or beverage to have an anti-diabetic effect when ingested.

11. Use of the Cinnamomum water based plant extract according to at least one of the claims 1 to 9 as a medicament.

12. Use of the Cinnamomum water based plant extract according to at least one of the claims 1 to 9 as an antioxidant.

13. Use of the Cinnamomum water based plant extract according to at least one of the claims 1 to 9 for the production of a medicament with hypoglycemic activity.

14. A composition comprising one or more plant extracts that are capable of delaying the onset and/or management of diabetes derived from the plant belonging to the plant sp. Cinnamomum.

15. A composition comprising one or more plant extracts, wherein the administration of the therapeutic composition can be formulated as pharmaceutical or naturopathic formulation such as phytoceuticals or nutraceuticals, for oral, topical, rectal, parenteral administration or for administration by inhalation or spray.

16. A composition comprising one or more plant extracts,, wherein the pharmaceutical or naturopathic formulations may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion hard or soft capsules, or syrups or elixirs.

17. A composition comprising one or more plant extracts, wherein the extracts can be formulated as phytoceuticals or nutraceuticals not limiting to teas, tonics, juices, syrups and nutraceuticals providing nutritional benefits in the form of foods, beverages, supplements and the like.

42