CLIAMS:I/We claim:
1. A composition comprising;
a. a xanthonoid; and
b. a flavonoid.
2. The composition as claimed in claim 1, wherein the xanthonoid is selected from the group consisting of tomentonone, zeyloxanthonone calozeyloxanthone, morellic acid, mangiferin, forbesione methylswertianin, bellidifolin, and combinations thereof, preferably mangiferin.
3. The composition as claimed in claim 1, wherein the flavonoid is selected from the group consisting of quercetin, kaempferol, luteolin, apigenin, catechin, hesperetin, naringenin, and combinations thereof, preferably catechin.
4. The composition as claimed in claim 1, wherein the w/w ratio of mangiferin to catechin is in the range of 80:20- 50:50, preferably 60:40, more preferably 50:50.
5. A food supplement comprising the composition as claimed in claim 1.
6. A method for management of Type 2 Diabetes Mellitus, wherein said method comprises oral administration to a subject in need thereof an effective amount of said food supplement as claimed in claim 5.
7. The method as claimed in claim 6, wherein oral administration can be in the form of a solid or a liquid.
8. The method as claimed in claim 6, wherein the subject is a mammal.
,TagSPECI:FIELD OF INVENTION
[0001] The present disclosure relates to a composition comprising of a xanthonoid, and a flavonoid and methods for management of Type II Diabetes Mellitus (T2DM).

BACKGROUND OF THE INVENTION
[0002] Type 2 Diabetes Mellitus (T2DM) is a global epidemic and a grave concern to developing country like India. One of the best known ways to maintain and manage T2DM is through life- style modifications, which includes changes in dietary habit. In this regard, it is pertinent to note that natural occurring ingredients play a pivotal role in delivering metabolic health benefits.
[0003] Mangiferin, a xanthone glucoside, is present in several plants such as Mangifera indica (mango), Anemarrhena asphodeloides (anemarrhena), Iris unguicularis (abington purple), Salacia reticulata (salacia), and Cyclopia subternata (honeybush). Mangiferin has been implicated in several beneficial biological effects including gastroprotective, immunomodulatory, lipolytic, anti-microbial, anti-inflammatory, anti-oxidant, and anti-diabetic effects. Several studies have shown that plant derived mangiferin has anti-hyperglycemic effects, and reduce oxidative stress (Raihan et al., Journal of Nutritional Therapeutics, 2013, 2, 74-79).
[0004] Catechin, a plant secondary metabolite belongs to the family of flavonoids and is a natural flavan-3-ol. In the nematode worm, Caenorhabditis elegans, catechin induces longevity (Saul et al., Mech. Ageing Dev., 2009, 130 (8), 477–86). Catechin is also well known for its antioxidant potential, and has been shown to inhibit the oxidation of low density lipoprotein and prevention of human plasma oxidation (Mangiapane et al., Biochem. Pharmacol., 1992, 43 (3), 445–450). In mice lacking ApoE protein, catechin has been shown to reduce atherosclerotic lesion development (Sylvain et al., Atherosclerosis, 2009, 204 (2), e21–27). In a human study, patients with T2DM who did not receive insulin therapy, catechin administration showed multiple benefits including prevention of obesity, recovery of insulin secretary ability and also helped to maintain low levels of glycosylated hemoglobin (Tomonori et al., Obesity, 2012, 15(6), 1473–1483).
[0005] Skeletal muscle cells take up the glucose and provide the substrate for the energy production thereby plays a major role in the whole body glucose homeostasis (Öberg et al., PLoS ONE, 2011 6(7), e22510). Skeletal muscles are one of the major insulin targets for blood glucose homeostasis. Glucose uptake defects in skeletal muscle cells are regularly seen in T2DM (Nedachi et al., Am J Physiol Endocrinol Metab., 2006, 291, E817–E828).
[0006] US7399491 describes a herbal fortified food product for promoting proven pharmacological activities, which comprises of mangiferin and catechin as plant extracts.
[0007] PCT/JP2009/066203 describes a composition comprising of mangiferin and catechin as components of plant extracts that function as antiallergenic agents or immunopotentiators.
[0008] Natural products with known beneficial effects apart from its intended function have fewer side effects in contrast to synthetic formulations and enjoy wider acceptance among the healthy and target population. Identifying such active bio-ingredients that can maintain glucose homeostasis is essential for delivering a holistic health benefit to the population dealing with T2DM.
[0009] Given the rising incidence of metabolic disorders such as diabetes among the population worldwide and increased awareness of natural products that combat such metabolic disorders, there is a need to develop high efficacy compositions that provide enhanced long-term management.

SUMMARY OF INVENTION
[0010] This summary is provided to introduce concepts related to a composition for management of T2DM comprising of a xanthonoid, and a flavonoid.
[0011] Yet another aspect of the present disclosure relates to a method for management of T2DM comprising administering an effective amount of the composition as described in the present disclosure.
[0012] Another aspect of the present disclosure relates to a food supplement comprising of a xanthonoid, and a flavonoid.
[0013] Yet another aspect of the present disclosure relates to a method for management of T2DM, said method comprising oral administration to a subject in need thereof an effective amount of a food supplement comprising Mangifera indica extract, and catechin.
[0014] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS
[0015] The following drawings form part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.
[0016] Figure 1 shows a graphical representation of the effect of ingredients such as diosgenin, capsaicin, genistein, catechin, and mangiferin on glucose uptake by murine C2C12 skeletal muscle cells, in accordance with an embodiment of the present disclosure.
[0017] Figure 2 shows a graphical representation of the dose response of mangiferin on glucose uptake by murine C2C12 skeletal muscle cells, in accordance with an embodiment of the present disclosure.
[0018] Figure 3 shows a graphical representation of the dose response of catechin on glucose uptake by murine C2C12 skeletal muscle cells, in accordance with an embodiment of the present disclosure.
[0019] Figure 4 shows a graphical representation of the dose response of a composition comprising mangiferin, and catechin in varying ratios on glucose uptake by murine C2C12 skeletal muscle cells, in accordance with an embodiment of the present disclosure.
[0020] Figure 5 shows a graphical representation of the synergistic effect of a composition comprising mangiferin, and catechin on glucose uptake by murine C2C12 skeletal muscle cells, in accordance with an embodiment of the present disclosure.

DETAILED DESCRPTION OF THE INVENTION
[0021] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure 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 such steps or features.
Definitions
[0022] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are collected here. These definitions should be read in the light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.
[0023] 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.
[0024] 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”.
[0025] 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.
[0026] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably.
[0027] As used herein, the term “management” does 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.
[0028] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference.
[0029] A composition comprising “synergistic activity” or a “synergistic composition” is a combination of compounds which exhibits increased biological or functional activity as a non-linear multiple of the biological or functional activity of the individual compounds. In other words, the combined biological or functional activity of two or more compounds being tested is significantly greater than the expected result based on independent effects of the compounds when tested separately. Synergy may be apparent only at some ranges or concentrations.
[0030] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein.
[0031] In an embodiment of the present disclosure, there is provided a composition for management of Type 2 Diabetes Mellitus, said composition comprising of a xanthonoid, and a flavonoid.
[0032] In an embodiment of the present disclosure, the xanthonoid is selected from the group consisting of tomentonone, zeyloxanthonone calozeyloxanthone, morellic acid, forbesione methylswertianin, manfigerin, and bellidifolin.
[0033] In a preferred embodiment of the present disclosure, the xanthonoid is mangiferin.
[0034] In an embodiment of the present disclosure, mangiferin is in the form of an extract from the bark of Mangifera indica.
[0035] In an embodiment of the present disclosure, the flavonoid is selected from the group consisting of quercetin, kaempferol, luteolin, apigenin, hesperetin, catechin, and naringenin.
[0036] In a preferred embodiment of the present disclosure, the flavonoid is catechin
[0037] In an embodiment of the present disclosure, mangiferin enhances the uptake of glucose by murine C2C12 skeletal muscle cells.
[0038] In an embodiment of the present disclosure, mangiferin concentration in the range of 1µg/mL to 50µg/mL enhances the uptake of glucose by murine C2C12 skeletal muscle cells.
[0039] In an embodiment of the present disclosure, catechin enhances the uptake of glucose by murine C2C12 skeletal muscle cells.
[0040] In an embodiment of the present disclosure, 10µg/mL catechin enhances the uptake of glucose by murine C2C12 skeletal muscle cells.
[0041] In an embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, said composition synergistically enhances the uptake of glucose by murine C2C12 skeletal muscle cells.
[0042] In an embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the w/w ratio of mangiferin to catechin is in the range of 80:20 to 50:50.
[0043] In an embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the w/w ratio of mangiferin to catechin is in the range of 60:40 to 50:50.
[0044] In a preferred embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the w/w ratio of mangiferin to catechin is 50:50.
[0045] In an embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the concentration of mangiferin in said composition is in the range of 5µg/mL to 8µg/mL.
[0046] In an embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the concentration of mangiferin in said composition is in the range of 6µg/mL to 8µg/mL.
[0047] In a preferred embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the concentration of mangiferin in said composition is 5µg/mL.
[0048] In an embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the concentration of catechin in said composition is in the range of 2µg/mL to 5µg/mL.
[0049] In a preferred embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the concentration of catechin is 5µg/mL.
[0050] In a preferred embodiment of the present disclosure, there is provided a composition comprising mangiferin, and catechin, wherein the concentration of mangiferin, and catechin is 5µg/mL and 5µg/mL respectively.
[0051] In an embodiment of the present disclosure, there is provided a food supplement comprising a composition consisting of mangiferin, and catechin for management of T2DM.
[0052] In an embodiment of the present disclosure, there is provided a food supplement comprising a composition consisting of mangiferin, and catechin for management of T2DM, wherein the w/w ratio of mangiferin to catechin is 80:20-50:50.
[0053] In an embodiment of the present disclosure, there is provided a food supplement comprising a composition consisting of mangiferin, and catechin for management of T2DM, wherein the w/w ratio of mangiferin to catechin is in the range of 60:40-50:50.
[0054] In a preferred embodiment of the present disclosure, there is provided a food supplement comprising a composition consisting of mangiferin, and catechin for management of T2DM, wherein the w/w ratio of mangiferin to catechin is 1:1.
[0055] In an embodiment of the present disclosure, there is provided a food supplement comprising a composition consisting of mangiferin, and catechin for management of T2DM, wherein the concentration of mangiferin, and catechin is 5µg/mL.
[0056] The composition of the present disclosure may be compounded with foods such as, but not limited to dairy products, grains, breads, meats, fruits, vegetables, rice and the like. The composition according to the present disclosure can be compounded with additional ingredients. For example, the composition can be mixed with carbohydrates lipids, polypeptides, fatty acids, phytochemicals, and combinations thereof.
[0057] Any conventional food processing technique may be used to achieve a product comprising the effective amount of the composition described herein. There is much information on the art and technology of the various conventional food processing techniques and their practices in both the pet food and food industries, and it is accordingly assumed that the general principals of these techniques are understood by the person skilled in the art.
[0058] In an embodiment of the present disclosure, the food supplement is in a solid form.
[0059] In an embodiment of the present disclosure, the food supplement is in a liquid form.
[0060] In an embodiment of the present disclosure, the food supplement is in a semi-solid form.
[0061] In an embodiment of the present disclosure, there is provided a method of management of T2DM, wherein said method comprises administering to a subject in need thereof a therapeutically effective amount of a composition comprising mangiferin, and catechin, wherein the effective amount is insufficient to suppress natural physiology at local or at a systemic level.
[0062] In another embodiment of the present disclosure, the subject in need thereof a therapeutically effective amount of the composition comprising a xanthonoid, and flavonoid is a mammal.
[0063] The present disclosure provides a useful composition for prevention and/or management of T2DM. The composition of the present disclosure comprises mangiferin, and catechin, wherein the composition is highly safe for consumption, useful and can be administered easily over a long term.
[0064] Although the subject matter has been described in considerable detail with reference to certain preferred embodiments thereof, other embodiments are possible.

EXAMPLES
[0065] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may vary.
Example 1
Preparation of solutions of test compounds used for screening for evaluation of glucose uptake
[0066] Preparation of mangiferin: mangiferin was sourced from Sigma (M3547). A 20.0 mg/mL stock solution was prepared by dissolving mangiferin powder in a solvent, preferably dimethyl sulfoxide (DMSO). A 1-50 µg/mL working solution of mangiferin was prepared by diluting stock solution in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, NY, USA).
[0067] Preparation of catechin: Catechin was sourced from Sigma (43412). A 10.0 mg/mL stock solution was prepared by dissolving catechin powder in a solvent, preferably DMSO. A 1 - 20 µg/mL working solution of catechin was prepared by diluting stock solution in DMEM (Gibco, NY, USA
[0068] Preparation of diosgenin: diosgenin was sourced from Sigma (D1634). A 8.3 mg/mL stock solution was prepared by dissolving diosgenin powder in a solvent, preferably dimethyl sulfoxide (DMSO). A 8.3 µg/mL working solution of diosgenin was prepared by diluting stock solution in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, NY, USA).
[0069] Preparation of capsaicin: capsaicin was sourced from Sigma (360376). A 7.6 mg/mL stock solution was prepared by dissolving capsaicin powder in a solvent, preferably dimethyl sulfoxide (DMSO). A 7.6 µg/mL working solution of capsaicin was prepared by diluting stock solution in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, NY, USA).
[0070] Preparation of genistein: genistein was sourced from Sigma (G6649). A 13.5 mg/mL stock solution was prepared by dissolving genistein powder in a solvent, preferably dimethyl sulfoxide (DMSO). A 13.5 µg/mL working solution of genistein was prepared by diluting stock solution in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, NY, USA).
Example 2
Cell culture and culture media
[0071] The cell culture media for C2C12 cells was prepared by adding 445mL of DMEM(Gibco, N.Y., USA) with 50mL Fetal Bovine Serum (FBS)(Invitrogen, NZ) and 5mL 100U/mL penicillin / 0.1mg/mL streptomycin (Sigma, USA). The solution was filtered through a 0.2 micron filter unit.
[0072] C2C12 murine skeletal muscle precursor cells (myoblasts) were obtained from American Type Culture Collection (Manassas, VA, USA) and maintained in proliferation media, consisting of DMEM supplemented with 10% FBS. Myoblasts were induced to differentiate into skeletal muscle myotubes by culturing for six-days in differentiation media, consisting of DMEM supplemented with 2% horse serum (HS) (Invitrogen, NZ). Differentiation media was replenished every 48 hours.
Example 3
Glucose uptake assay
[0073] The C2C12 cells were allowed to get 70% confluent before differentiation was initiated. The C2C12 cells were treated with DMEM supplemented with 2 % (v/v) HS. The cells were subsequently re-fed every 48 hours with 2% HS supplemented DMEM.
[0074] At the 6th day of differentiation, the culture media were removed and replaced with serum-free DMEM containing no glucose and incubated at 37 °C with 5% CO2 for one-hour. The C2C12 cells were subsequently re-fed for one-hour with serum free no glucose DMEM with or without treatments. Subsequently, serum free media with or without treatment were removed and cells were incubated at 37 °C with 5% CO2 for 30 minutes with 250µM of fluorescent analogue of glucose, 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (6-NBDG) in serum free no glucose DMEM. The 6-NBDG uptake reaction was stopped by removing the incubation medium and washing the cells twice with pre-chilled phosphate buffered saline (PBS pH 7.4). Cells in each well were subsequently re-suspended in pre-chilled fresh Opti-MEM (Invitrogen, NZ) and maintained at 4°C for fluorimetry analysis performed within 30 minutes. The experiments were conducted in 96-well plates. All the treatment groups were done in triplicates.
Example 4
Effect of various compounds on glucose uptake by murine C2C12 skeletal muscle cells
[0075] Figure 1 shows the effect of various compounds found in natural extracts on glucose uptake by murine C2C12 skeletal muscle cells. It can be inferred from Figure 1 that catechin, and mangiferin both, are able to significantly enhance the uptake of glucose by murine C2C12 skeletal muscle cells compared to the other compounds.
Example 5
Dose response of mangiferin on glucose uptake by murine C2C12 skeletal muscle cells
[0076] Figure 2 shows the effect of varying dose of mangiferin on glucose uptake by murine C2C12 skeletal muscle cells. It can be inferred from Figure 2 that mangiferin dose ranging from 1µg/mL to 50µg/mL can effectively enhance glucose uptake by C2C12 cells, but there are no significant differences in the effect between the spectrums of doses tested. For further experiments, a lower value of 10µg/mL was used.
Example 6
Dose response of catechin on glucose uptake by murine C2C12 skeletal muscle cells
[0077] Figure 3 shows the effect of varying dose of catechin on glucose uptake by murine C2C12 skeletal muscle cells. It can be inferred from Figure 3 that 10µg/mL dose of catechin is able to significantly enhance glucose uptake by C2C12 cells compared to lower or higher doses as tested.
Example 7
Effect of a combination of mangiferin, and catechin on glucose uptake by murine C2C12 skeletal muscle cells
[0078] Mangiferin and catechin combination in varing w/w ratios were tested for their effect on glucose uptake by C2C12 cells. The various w/w ratios are listed in Table 1.
Table 1
Ratio (w/w) Ingredient concentrations (µg/mL)
Mangiferin : Catechin Mangiferin Catechin
100:0 10 0
80:20 8 2
60:40 6 4
50:50 5 5
40:60 4 6
20:80 2 8
0:100 0 10

[0079] Figure 4 shows the effect of various w/w ratios of mangiferin and catechin combinations on glucose uptake by murine C2C12 skeletal muscle cells. It can be inferred from Figure 4 that a synergistic effect can be observed over a broad range of 80:20(mangiferin:catechin) to 50:50(mangiferin:catechin).this is surprising as at these ratios, the combined effect is more than an additive response compared to responses of each component individually at same concentrations (Table 1).
Example 8
Synergistic effect of a 50:50 w/w ratio of mangiferin to catechin on glucose uptake by C2C12 murine skeletal muscle cells
[0080] Figure 5 shows the maximal synergistic effect of a composition comprising mangiferin and catechin. The w/w ratio of amngiferin to catechin is 1:1. The concentration of mangiferin,and catechin in aforementioned composition is 5µg/mL. it can be inferred from Figure 5 that compared to the effect of 5µg/ml mangiferin or 5µg/mL catechin alone, the effect of the combination of mangiferin and catechin at same concentrations (5µg/mL each) has an effect that is more than twice of expected additive effect.