FIELD OF INVENTION
[001] The present disclosure broadly relates to an anti-diabetic composition and particularly relates to the anti-diabetic composition comprising herbal extracts. The present disclosure further relates to a process of preparing the composition.
BACKGROUND OF INVENTION
[002] Diabetes mellitus is a common and a serious disease that is one of the major health concern all over the world. The number of people affected by diabetes mellitus are increasing tremendously. According to World Health Organization (WHO), more than 190 million people are suffering from diabetes mellitus (International Diabetes Federation 2004 - International Association for the Study of Obesity, Brussels, 2004. (Online) 2004 (Cited 2004 June 25))
[003] Diabetes mellitus is a chronic disorder that is characterized by elevated concentration of blood glucose or hyperglycemia. Diabetes mellitus is either caused by decreased secretion of insulin by the pancreas or resistance to the action of insulin in various tissues (e.g. muscle, liver, adipose) uptake. Elevated glucose levels, in turn, often leads to secondary health problems that require additional medical treatment. Some of the leading diabetes-related health risks include hyperglycemia, arteriosclerosis, diabetic retinopathy (possibly leading to blindness), cataracts, nephropathy, increased risk of infections, hypertension, nerve disease, risk of amputations, impotence, diabetic ketoacidosis, and dementia (Geiss LS, Herman WH, Smith PJ. Mortality in non-insulin-dependent diabetes. In: Harris MI, Cowie CC, Stern MP, et al, eds. Diabetes in America. 2nd ed. Bethesda, MD: National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases; 1995:233q257. NIH publication 95-1468)
[004] Primarily, there are two types of Diabetes mellitus, i.e. type 1 (insulin dependent diabetes mellitus; IDDM) and type II (non- insulin-dependent diabetes mellitus; NIDDM). Type 1 diabetes generally occurs in childhood and results from the body's inability to produce insulin. Type 2 diabetes is the more prevalent form and results from either insulin deficiency or, more commonly, from insulin resistance, accounting for more than 90% of patients. Some diabetic patients are able

to maintain healthy blood glucose concentrations with restrictive diets and increase
exercise routine. However, people with diabetes mostly require the assistance of oral
hypoglycemic drugs and/or insulin replacement therapy by injection.
[005] US Patent Application US6011049A discloses a composition of a glitazone
antidiabetic agent and a biguanide antidiabetic agent# for treating diabetes mellitus
and improving glycemic control.
[006] US Patent Application US5859051A discloses antidiabetic agents. The
corresponding Patent Application further relates to compositions and methods for
the use of the compounds in the treatment of diabetes and obesity and for lowering
or modulating triglyceride levels and cholesterol levels or raising high density
lipoprotein levels or for increasing gut motility or for treating atherosclerosis are also
disclosed.
[007] However, the potential use of conventional hypoglycemic agents (such as
sulfonylureas, biguanides and alpha glucosidase inhibitors) and insulin injections
that provides symptomatic relief are associated with various side effects. Further,
drug therapy involving these hypoglycemic agents and insulin injections, do not
always achieve satisfactory glycemic control. Moreover, these conventional
treatments often lower blood glucose by promoting the non-selective uptake of
glucose by many cells, including adipocytes, which contributes to undesirable
weight gain. Therefore, continuous treatment modifications and monitoring are often
necessary with the use of the conventional hypoglycemic agents Additionally,
traditional treatments, like insulin injections, are expensive and can be painful to
administer.
[008] Accordingly, due to the ongoing problems with regards to the use of the
conventional treatments or composition for treatment of diabetes, there remains a
need in the art to develop an alternative composition which would be devoid of the
above-mentioned problems associated with conventional treatment method.
SUMMARY OF THE INVENTION
[009] In an aspect of the present disclosure, there is provided an anti-diabetic
composition comprising: (a) Silybum marianum extract; and (b) Ginkgo biloba

extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1.
[0010] In an aspect of the present disclosure, there is provided a process for preparing the anti-diabetic composition comprising: (a) Silybum marianum extract; and (b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, said process comprising: (i) obtaining a first mixture comprising Silybum marianum extract; (ii) obtaining a second mixture comprising Ginkgo biloba extract; and (iii) contacting the first mixture to the second mixture, to obtain the anti-diabetic composition. [0011] In an aspect of the present disclosure, there is provided a process for preparing the anti-diabetic composition comprising: (a) Silybum marianum extract; (b) Ginkgo biloba extract; and c) at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein (SMP), malt extract, flavour, masking agent, and combinations thereof, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, said process comprising: i) obtaining a first mixture comprising Silybum marianum extract; ii) obtaining a second mixture comprising Ginkgo biloba extract; iii) obtaining the at least one excipient; and iv) contacting the first mixture, the second mixture to the at least one excipient, to obtain the anti-diabetic composition. [0012] 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 ACCOMPANYING DRAWINGS
0013] The following drawings form a part of the present specification and are ncluded 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.

[0014] Figure 1 depicts the effect of varying concentrations of Silybum marianum extract and Ginkgo biloba extract on glucosidase inhibition, in accordance with an embodiment of the present disclosure.
[0015] Figure 2 depicts the glucosidase inhibitory effect of combination of Silybum marianum extract and Ginkgo biloba extract at various concentrations, in accordance with an embodiment of the present disclosure
DETAILED DESCRIPTION OF THE INVENTION
[0016] 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
[0017] For convenience, before further description of the present disclosure, certain
terms employed in the specification, and examples are delineated 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.
[0018] 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.
[0019] 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".
[0020] 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.

[0021] The term "including" is used to mean "including but not limited to". "Including" and "including but not limited to" are used interchangeably. [0022] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.
[0023] Silybum marianum is an annual or biennial herb that belongs to the family Asteraceae. The common name of Silybum marianum is Milk thistle. This plant is widely used as an alternative medicine that has been considered an excellent medical remedy since the time of ancient Greece. This plant has been used as a natural extract for liver diseases. The main active ingredients are alkaloids, flavonoids, saponin, tannin, phenolic compounds and many more collectively known as silymarin. Silymarin is commercially available as standardized extract. Silymarin exhibits antioxidant properties. Additionally, silymarin helps in inhibiting lipid peroxidation, prevents glutathione depletion, and activates antioxidant enzymes that protect DNA from degradation. Therefore, Silybum marianum is known for treating various diseases and it does not cause any significant side effect (Silymarin in Type 2 Diabetes Mellitus: A Systematic Review and Meta-Analysis of Randomized Controlled Trials, Luminita Voroneanu, Ionut Nistor, Raluca Dumea, Mugurel Apetrii, and Adrian Covic).
[0024] Ginkgo biloba has been used in traditional Chinese medicine for 5000 years. Gingko biloba extract is derived from leaves of Gingko biloba. It is the only surviving species of the Ginkgoaceae family. Ginkgo biloba extract is popularly used a dietary supplement that is ingested by the general population to enhance mental focus and to delay onset of age-acquired loss of cognitive function by elders. Ginkgo biloba extract helps in decreasing the efficacy of the hypoglycemic agents and further increasing the whole-body insulin resistance. Thereby, it is known to be an alternative medication for treating patients with non-insulin dependent diabetes (NIDDM) (Ginkgo biloba Extract for Patients with Early Diabetic Nephropathy: A

Systematic Review, Lei Zhang, Wei Mao, Xinfeng Guo, Yifan Wu, Chuang Li, Zhaoyu Lu, Guobin Su, Xiaoyan Li, Zhuangzhu Liu, Rong Guo, Xina Jie, Zehuai Wen and Xusheng Liu).
[0025] As used herein, alpha-glucosidase enzyme is a glucosidase enzyme that is present in the brush border of the small intestine. Alpha-glucosidase bonds breaks down starch and disaccharides to glucose by acting upon a(l-4) bonds. Inhibition of alpha-glucosidase enzyme helps to prevent the fast breakdown of sugars and thereby controlling the blood sugar level. Therefore, inhibition of alpha-glucosidase enzyme plays a pivotal role in treating diabetes.
[0026] For the purposes of the present document, the term "synergistic effect" or "synergy" is used to describe the interaction or cooperation of two or more organizations, substances, or other agents to produce a combined effect greater than the sum of their separate effects.
[0027] For the purposes of the present document, vitamin premix refers to vitamins selected from the group consisting of Vitamin A, D, E, K, Vitamin B complex, and combinations thereof. Mineral premix refers to minerals selected from the group consisting of sodium, magnesium, calcium, chloride, iron, and combinations thereof, [0028] For the purposes of the present document, malt extract refers to the processed forms of malt selected from the group consisting of liquid extract, dry extract and combinations thereof. Masking agent refers to flavour ingredients that have little or no taste or smell of their own but complement, enhance or otherwise modify the flavour of a food product.
[0029] As disclosed in the background section of the present disclosure, various conventional treatments have been deployed for treating diabetes mellitus. Conventional methods of treatments such as injections and hypoglycemic agents that are widely used for alleviating, preventing or treating symptoms of diabetes are associated with various side effects. Particularly, the excessive use of insulin injections for treating diabetes carries a high risk of low blood sugar reactions. and further contributing to undesirable weight gain. The use of insulin injections and insulin pumps requires continuous monitoring. Additionally, the underlying cost of

these conventional treatments is becoming too expensive for many diabetic patients to afford
[0030] To overcome the problems faced by the conventional treatment of diabetes, the present disclosure deploys the use of a natural, plant-based, nontoxic and herbal preparation. The object of the present disclosure is to provide a combination of plant constituents such as Silybum marianum extract and Ginkgo biloba extract that results in obtaining effective and safe anti-diabetic composition. Additionally, the use of combination of Silybum marianum extract and Ginkgo biloba extract in a form of tablets, capsules, or any other dosage forms, that shows enhanced alpha-glucosidase inhibition, can be utilized to deliver effective and safe anti-diabetic composition. [0031] 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.
[0032] 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. [0033] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1.
[0034] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1. In another embodiment of the present disclosure, the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 8:1 to 0.50:1. In yet another embodiment of the present disclosure, the Silybum

marianum extract to the Ginkgo biloba extract weight ratio is in a range of 4:1 to 0.6:1.
[0035] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1. In another embodiment of the present disclosure, the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 7:1 to 0.60:1.
[0036] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, and wherein the Silybum marianum extract has a weight percentage in a range of 0.005-0.05% with respect to the anti-diabetic composition. In another embodiment of the present disclosure, the Silybum marianum extract has a weight percentage in a range of 0.01-0.04% with respect to the anti-diabetic composition.
[0037] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, and wherein the Ginkgo biloba extract has a weight percentage in a range of 0.002-0.02% with respect to the anti-diabetic composition. [0038] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the Silybum marianum extract has a weight percentage in a range of 0.005-0.05% with respect to the anti-diabetic composition and wherein the Ginkgo biloba extract has a weight percentage in a range of 0.002-0.02% with respect to the anti-diabetic composition.
[0039] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is

in a range of 10:1 to 0.50:1 and wherein the Silybum marianum extract comprises silymarin having a weight percentage in a range of 80-90% with respect to the Silybum marianum extract. In another embodiment of the present disclosure, the Silybum marianum extract comprises silymarin having a weight percentage in a range of 83-90% with respect to the Silybum marianum extract. [0040] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, and wherein the Ginkgo biloba extract comprises flavonoids having a weight percentage in a range of 24-30% with respect to the Ginkgo biloba extract. In another embodiment of the present disclosure, the Ginkgo biloba extract comprises flavonoids having a weight percentage in a range of 27-30% with respect to the Ginkgo biloba extract.
[0041] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the Silybum marianum extract comprises silymarin having a weight percentage in a range of 80-90% with respect to the Silybum marianum extract and wherein the Ginkgo biloba extract comprises flavonoids having a weight percentage in a range of 24-30% with respect to the Ginkgo biloba extract
[0042] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the Silybum marianum extract has a weight percentage in a range of 0.005-0.05% with respect to the anti-diabetic composition and wherein the Ginkgo biloba extract has a weight percentage in a range of 0.002-0.02% with respect to the anti-diabetic composition and wherein the Silybum marianum extract comprises silymarin having a weight percentage in a range of 80-90% with respect to the Silybum marianum extract and wherein the Ginkgo biloba

extract comprises flavonoids having a weight percentage in a range of 24-30% with respect to the Ginkgo biloba extract
[0043] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the composition further comprises at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, emulsifier, maltodextrin, and combinations thereof. In another embodiment of the present disclosure, the flavor includes, but are not limited to vanilla and chocolate and combinations thereof. [0044] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the composition further comprises at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, emulsifier, maltodextrin, and combinations thereof. In another embodiment of the present disclosure, the masking agent include, but are not limited to vanilla micron, chocolate micron and combinations thereof.
[0045] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, c) at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, emulsifier, maltodextrin, and combinations thereof, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 7:1 to 0.60:1. [0046] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, c) at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, emulsifier, maltodextrin, and combinations thereof, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the

Silybum marianum extract has a weight percentage in a range of 0.005-0.05% with respect to the anti-diabetic composition and wherein the Ginkgo biloba extract has a weight percentage in a range of 0.002-0.02% with respect to the anti-diabetic composition
[0047] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, c) at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, emulsifier, maltodextrin, and combinations thereof, wherein the Silybum marianum extract comprises silymarin having a weight percentage in a range of 80-90% with respect to the Silybum marianum extract and wherein the Ginkgo biloba extract comprises flavonoids having a weight percentage in a range of 24-30% with respect to the Ginkgo biloba extract
[0048] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, c) at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, emulsifier, maltodextrin, and combinations thereof, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the Silybum marianum extract has a weight percentage in a range of 0.005-0.05% with respect to the anti-diabetic composition and wherein the Ginkgo biloba extract has a weight percentage in a range of 0.002-0.02% with respect to the anti-diabetic composition, and wherein the Silybum marianum extract comprises silymarin having a weight percentage in a range of 80-90% with respect to the Silybum marianum extract and wherein the Ginkgo biloba extract comprises flavonoids having a weight percentage in a range of 24-30% with respect to the Ginkgo biloba extract.
[0049] In an embodiment of the present disclosure, there is provided a process for preparing the anti-diabetic composition comprising (a) Silybum marianum extract; and (b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, said process comprising: (i) obtaining a first mixture comprising Silybum marianum extract; (ii) obtaining a

second mixture comprising Ginkgo biloba extract; and (iii) contacting the first mixture to the second mixture, to obtain the anti-diabetic composition. In another embodiment of the present disclosure, the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 7:1 to 0.60:1.
[0050] In an embodiment of the present disclosure, there is provided a process for preparing the anti-diabetic composition comprising (a) Silybum marianum extract; (b) Ginkgo biloba extract; c) at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, and combinations thereof, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, said process comprising: i) obtaining a first mixture comprising Silybum marianum extract; ii) obtaining a second mixture comprising Ginkgo biloba extract; iii) obtaining the at least one excipient; and iv) contacting the first mixture, the second mixture to the at least one excipient, to obtain the anti-diabetic composition. In another embodiment of the present disclosure, the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 7:1 to 0.60:1.#
[0051] In an embodiment of the present disclosure, there is provided an anti-diabetic composition comprising: a) Silybum marianum extract; and b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1 and wherein the composition inhibits production of alpha-glucosidase enzyme.
[0052] In an embodiment of the present disclosure, there is provided a process for preparing the anti-diabetic composition comprising (a) Silybum marianum extract; and (b) Ginkgo biloba extract, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, said process comprising: (i) obtaining a first mixture comprising Silybum marianum extract; (ii) obtaining a second mixture comprising Ginkgo biloba extract; and (iii) contacting the first mixture to the second mixture, to obtain the anti-diabetic composition [0053] In an embodiment of the present disclosure, there is provided a process for preparing the anti-diabetic composition comprising (a) Silybum marianum extract; (b) Ginkgo biloba extract; c) at least one excipient selected from a group consisting

of vitamin premix, mineral premix, solid milk protein, malt extract, flavor, masking agent, and combinations thereof, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1, said process comprising: i) obtaining a first mixture comprising Silybum marianum extract; ii) obtaining a second mixture comprising Ginkgo biloba extract; iii) obtaining the at least one excipient; and iv) contacting the first mixture, the second mixture to the at least one excipient, to obtain the anti-diabetic composition and wherein the composition inhibits production of alpha-glucosidase enzyme. [0054] In an embodiment of the present disclosure, there is provided the anti-diabetic composition as described herein, wherein the composition is in a form of capsules, tablets, powder and other dosage forms.
[0055] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[0056] 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 apply.
[0057] The working and non-working examples as depicted in the forthcoming sections highlight the criticality of the working percentages of different ingredients in achieving the anti-diabetic composition of the present disclosure. It is specified that the use of combination of individual extracts such as Silybum marianum extract and Ginkgo biloba extract at specific weight ratio range and specific weight

percentage range shows synergistic effect in inhibiting the alpha Glucosidase enzyme. It is further specified that the absence of any of the plant ingredients mentioned above or their presence beyond the specific weight ratio range and weight percentage ranges as disclosed herein fails to achieve the desirable effect on inhibition of alpha glucosidase enzyme.
EXAMPLE 1
Inhibitory effect on alpha glucosidase enzyme displayed by individual extracts [0058] The alpha glucosidase enzyme inhibitory activity of individual extract concentration of Silybum marianum extract and Ginkgo biloba extract was determined by using alpha glucosidase enzyme.
Materials and Method
[0059] The individual extracts such as Silybum marianum extract and Ginkgo biloba extract used for arriving the anti-diabetic composition of the present disclosure were commercially procured. The commercially procured Silybum marianum extract consists of silymarin that is having a weight percentage in a range of 80-90% with respect to the anti-diabetic composition. The commercially procured Ginkgo biloba extract consists of flavonoids that is having a percentage in a range of 24-30% with respect to the anti-diabetic composition.
Optimization of individual extract concentration using alpha glucosidase assay [0060] The alpha glucosidase enzyme inhibitory activity of individual concentration of Silybum marianum extract and Ginkgo biloba extract was determined by using alpha glucosidase assay. For performing the alpha-glucosidase assay,10µl of individual extract was taken. 100µl of alpha-glucosidase enzyme (1µl/ml) was added to 10µl of individual extract to form the reaction mixture. The reaction mixture was then allowed to incubate for 10 minutes. After incubating for 10 minutes, 50µl of 5mM p-nitro phenyl-alpha glucopyranoside solution was added to the reaction mixture and was further allowed to incubate for 5 minutes. Absorbance was read at 405nm.

Percentage inhibition of alpha-glucosidase enzyme of the test sample was calculated by using the formula:
%inhibition = [Absorbance ctrl- Absorbance test]/ Absorbance ctrl * 100 [0061] Individual extracts such as Silybum marianum extract and Ginkgo biloba extract were used for preparing the anti-diabetic composition. The effect of Silybum marianum extract and Ginkgo biloba extract on inhibition of alpha glucosidase enzyme were studied individually at various concentrations. [0062] Assessing the data in Figure 1, the effect on alpha-glucosidase enzyme was derived for Silybum marianum extract and Ginkgo biloba extract individually at various concentrations. The results obtained in the figure 1, represents + SD. [0063] Referring to Figure 1, it was observed that the maximum inhibition of alpha-glucosidase enzyme was observed by using Ginkgo biloba extract at a concentration of 12.5mg/ml. Similarly, when Silybum marianum extract was used individually at a concentration of 1mg/ml, alpha-glucosidase enzyme was maximally inhibited by 35%.
EXAMPLE 2
Various formulations comprising the composition of the present disclosure
Formulation 1 [0064] The formulation as disclosed in table 1 was prepared by: i) adding individual extracts such as Silybum marianum extract (40mg), Ginkgo biloba extract (3.6mg); and excipients such as vitamin premix (100mg), mineral premix (1g), SMP (25g), malt extract (25g) and other excipients combined such as 48.85 gram of flavours (e.g.: vanilla, chocolate), masking agents (e.g.: vanilla micron, chocolate micron), emulsifier, maltodextrin; and ii) Mixing all ingredients at room temperature, to obtain a homogeneous powder providing the final formulation comprising all the ingredients as depicted in table 1.

The formulation 2 was prepared in the similar manner as Formulation 1. However, the formulation 2 contained different amount of individual extracts such as Silybum marianum extract (30mg), Ginkgo biloba extract (7.2mg), excipients such as vitamin premix (100mg), mineral premix (1g), SMP (25g), malt extract (25g) and other excipients combined such as 48.86 gram of flavours (e.g.: vanilla, chocolate), masking agents (e.g.: vanilla micron, chocolate micron), emulsifier, maltodextrin. Table 2 depicts the formulation 2.

The formulation 3 was prepared in the similar manner as Formulation 1. However, the formulation 3 contained different amount of individual extracts such as Silybum marianum extract (25mg), Ginkgo biloba extract (9.0mg), excipients such as vitamin premix (100mg), mineral premix (1g), SMP (25g), malt extract (25g) and other excipients combined such as 48.86 gram of flavours (e.g.: vanilla, chocolate), masking agents (e.g.: vanilla micron, chocolate micron), emulsifier, maltodextrin. Table 3 depicts the formulation 3.
The formulation 4 was prepared in the similar manner as Formulation 1. However, the formulation 4 contained different amount of individual extracts such as Silybum marianum extract (20mg), Ginkgo biloba extract (10.8mg), excipients such as vitamin premix (100mg), mineral premix (1g), SMP (25g), malt extract (25g) and other excipients combined such as 48.86 gram of flavours (e.g.: vanilla, chocolate), masking agents (e.g.: vanilla micron, chocolate micron), emulsifier, maltodextrin. Table 4 depicts the formulation 4.

The formulation 5 was prepared in the similar manner as Formulation 1. However, the formulation 5 contained different amount of individual extracts such as Silybum marianum extract (10mg), Ginkgo biloba extract (14.4mg), excipients such as vitamin premix (100mg), mineral premix (1g), SMP (25g), malt extract (25g) and other excipients combined such as 48.87 gram of flavours (e.g.: vanilla, chocolate), masking agents (e.g.vanilla micron, chocolate micron) , emulsifier, maltodextrin. Table 5 depicts the formulation 5.

[0069] The anti-diabetic composition of the present disclosure was prepared by mixing Silybum marianum extract and Ginkgo biloba extract in specific weight percentages and ratios as described in the present disclosure. The anti-diabetic composition is utilized for inhibiting alpha-glucosidase enzyme. Alpha-glucosidase assay was performed to assess the inhibitory effect of Silybum marianum extract and Ginkgo biloba extract on alpha-glucosidase enzyme. [0070] Table 6 depicts the ratios and concentrations of Silybum marianum extract and Ginkgo biloba extract. The concentrations of Silybum marianum extract and Ginkgo biloba extract were used in combination at different ratio to study the inhibitory effect of Silybum marianum extract and Ginkgo biloba extract on alpha-glucosidase enzyme.

[0071] Table 7 depicts the working examples (WE) 1 to 4. Table 7 further depicts various concentration of Silybum marianum extract and Ginkgo biloba extract that were showing synergistic effect on inhibition of alpha glucosidase enzyme. [0072] The Working and Non-Working Examples as depicted in the present Examples were obtained by mixing the individual components as per the respective concentrations depending upon the specific Example. The individual components were blended together at room temperature to obtain dry powder. [0073] For experimental purpose, solvents were used for preparing individual components. As per the present disclosure, solvents include polar and non-polar solvents, specifically, dimethyl sulfoxide (DMSO) was used for preparing the solutions of respective components.
[0074] Referring to Figure 2, it was observed that the combination of Silybum marianum extract and Ginkgo biloba extract used at a concentration of 0.15mg/ml and 0.036mg/ml in WE 1 was showing synergistic effect, that was inhibiting the alpha glucosidase enzyme by 8%. Similarly, WE 2 and WE 3 shows enhanced alpha-glucosidase enzyme inhibition, when Silybum marianum extract and Ginkgo biloba extract were used at a concentration of 0.125mg/ml (Silybum marianum extract) and 0.045mg/ml (Ginkgo biloba extract), 0.1mg/ml (Silybum marianum extract) and 0.054mg/ml (Ginkgo biloba extract) respectively in WE 2 and WE 3. It is further analysed from Figure 2 and Figure 1, that the inhibitory effect on alpha glucosidase enzyme of 11% by using the combination of 0.05mg/ml Silybum marianum extract and 0.072mg/ml Ginkgo biloba (WE 4), is more than the additive effect of the alpha glucosidase enzyme inhibition (approximately at 6%) by using said combination of 0.05mg/ml Silybum marianum extract and 0.072mg/ml Ginkgo biloba, thereby affirming the synergistic effect of the combination.

[0075] Table 8 depicts the non-working example (NWE) 1-4. It was observed that the NWE-1, NWE-2, NWE-3 and NWE-4 were not showing any synergistic effect on alpha-glucosidase enzyme as the weight ratios of NWE-1, NWE-2, NWE-3 and NWE-4 were falling outside the working ranges as disclosed in the present disclosure. It was observed in NWE-1, that by using Silybum marianum extract and Ginkgo biloba extract at a concentration of 0.2mg/ml and 0.018mg/ml, the combination of Silybum marianum extract and Ginkgo biloba extract was not showing the synergistic effect
In case of NWE-2, the weight ratio of Silybum marianum extract and Ginkgo biloba was 0.427:1 that is falling outside the range of weight ratio 10:1 to 0.5:1, as disclosed in the present disclosure. Similarly, in case of NWE-3 and NWE-4, the weight ratio of Silybum marianum extract and Ginkgo biloba were 0.16:1 and 0.328:1 respectively, that is falling outside the range of weight ratio 10:1 to 0.5:1 of the present disclosure.

[0076] Table 9 depicts the Non-Working Examples that uses Silybum marianum extract with Rosemary extract instead of Ginkgo biloba extract. It was further observed that none of the non-working examples (NWE) 5-9 mentioned in the table 9 are showing synergistic effect on inhibition of alpha-glucosidase enzyme. Therefore, it can be inferred that the absence or replacement of any one of the critical ingredient with any other ingredient fails to yield any desirable effect.

[0077] Table 10 depicts the Non-Working Examples that uses Ginkgo biloba extract with Hops extract instead of Silybum marianum extract. It can be observed that that all of the non-working examples (NWE) 10-14 mentioned in the table 10 failed to show any synergistic effect on inhibition of alpha-glucosidase enzyme. Therefore, it can be appreciated that replacing any one of the critical ingredient with any other ingredient does not yield any desirable effect.
[0078] Overall, the combination of Silybum marianum extract and Ginkgo biloba in the disclosed weight ratio and percentage ranges is essential to have a synergistic effect on alpha-glucosidase enzyme inhibition. Replacing even a single plant ingredient with another ingredient does not provide the desired effect. Also, deviating from the disclosed weight ratio and weight percentage ranges does not exhibit the desired result.
Advantages of the present disclosure
[0079] The present disclosure discloses an anti-diabetic composition comprising Silybum marianum extract and Ginkgo biloba extract, wherein Silybum marianum extract to Ginkgo biloba extract are present as per the disclosed weight ratio range. The anti-diabetic composition shows synergistic effect, that provides enhanced inhibition of alpha glucosidase enzyme in natural ways. Additionally, the raw

materials for preparing the anti-diabetic composition are readily available that effectively treats diabetes without inducing any undesirable side effects. Due to its affordable cost and safety profile, the anti-diabetic composition is considered as an effective treatment of diabetes.

I/We Claim:
1. An anti-diabetic composition comprising:
a) Silybum marianum extract; and
b) Ginkgo biloba extract,
wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 10:1 to 0.50:1.
2. The anti-diabetic composition as claimed in claim 1, wherein the Silybum marianum extract to the Ginkgo biloba extract weight ratio is in a range of 7:1 to 0.60:1.
3. The anti-diabetic composition as claimed in claim 1, wherein the Silybum marianum extract has a weight percentage in a range of 0.005-0.05% with respect to the anti-diabetic composition.
4. The anti-diabetic composition as claimed in claim 1, wherein the Ginkgo biloba extract has a weight percentage in a range of 0.002-0.02% with respect to the anti-diabetic composition.
5. The anti-diabetic composition as claimed in claim 1, wherein the Silybum marianum extract comprises silymarin having a weight percentage in a range of 80-90% with respect to the Silybum marianum extract.
6. The anti-diabetic composition as claimed in claim 1, wherein the Ginkgo biloba extract comprises flavonoids having a weight percentage in a range of 24-30% with respect to the Ginkgo biloba extract.
7. The anti-diabetic composition as claimed in claim 1, wherein the composition further comprises at least one excipient selected from a group consisting of vitamin premix, mineral premix, solid milk protein (SMP), malt extract, flavor, masking agent, and combinations thereof.
8. A process for preparing the anti-diabetic composition as claimed in claim 1, said process comprising:

a) obtaining a first mixture comprising Silybum marianum extract;
b) obtaining a second mixture comprising Ginkgo biloba extract; and
c) contacting the first mixture to the second mixture, to obtain the anti-diabetic composition.

9. A process for preparing the composition as claimed in claim 7, said process
comprising:
a) obtaining a first mixture comprising Silybum marianum extract;
b) obtaining a second mixture comprising Ginkgo biloba extract;
c) obtaining the at least one excipient; and
d) contacting the first mixture, the second mixture to the at least one excipient, to obtain the anti-diabetic composition.
10. The anti-diabetic composition as claimed in claim 1, wherein the composition
inhibits production of alpha-glucosidase enzyme.