TECHNICAL FIELD
[001] The subject matter described herein in general relates to prebiotic compositions, and in particular relates to prebiotic compositions of plant origin for improving gut health.
BACKGROUND OF INVENTION
[002] External agents like radiations, microbes, pollutants and sun light triggers the generation of free radicals (ROS) in the body which leads to oxidative stress. Overproduction of ROS affects the oxidative modification of lipids, proteins, nucleic acids (DNA damage), inflammation and tissue damage. This causes the imbalance of natural antioxidative system, and further leads to several health problems like premature aging, cellular apoptosis, heart failure, cancer and neuro-degeneration (M. M. Towhidul Islam et.al 2014 Free Radicals in Human Health and Disease pp 59-73.) Based on the literature, consumption of plant extracts or plant derived molecules provides the protection against oxidative stress which is caused by external agents (Pallavi Sharma, et.al Journal of Botany 2012,26; Aherne SA et.al. British Journal of Nutrition (2007), 97, 321s328.)
[003] Recent studies show that herbal extracts have a good prebiotic potential to restore gastrointestinal microflora. For example, US20120269790 discloses the use of olive (Olea europaea) and pomegranate (Punica granatum) plant extracts as prebiotics. WO2015008139 discloses a composition comprising Perilla preparation for use in promoting growth of probiotic bacteria in a subject as a prebiotic and/or for use in inhibiting the growth of pathogenic bacteria selected from the group consisting of Enterobacteriaceae, Bacteroides and/or Clostridia in a subject. WO2014160476 discloses a prebiotic composition containing a fructan and ginger root extract, among other ingredients for growth of probiotic bacteria, and to also help improve the health of the individual.
[004] Although literature discloses the use of prebiotics compositions in food supplements and personal care products, there still exists a need to develop novel prebiotic compositions that can impart additional physiological benefits, such as anti-oxidant and anti-microbial activity.

SUMMARY OF THE INVENTION
[005] In an aspect of the present invention, there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1.
[006] In another aspect of the present invention, there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; and iii) contacting the Cinnamomum zeylanicum extract, and the Glycyrrhiza glabra extract, to obtain the composition. [007] 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 DRAWINGS
[008] The detailed description is described with reference to the accompanying figures. The same numbers are used throughout the drawings to reference like features and components.
[009] Figure 1 depicts the effect of Cinnamomum zeylanicum extract (cinnamon); and Glycyrrhiza glabra extract (licorice), alone, and in combination at a weight ratio of 10:1 (cinnamon to licorice), on the probiotic bacteria (Bacillus subtilis CNCM I-2745) and pathogen (E. coli MTCC448), in accordance with an implementation of the present subject matter.
[0010] Figure 2 depicts the effect of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, alone, and in combination at varying weight ratios, on nitric oxide (NO) inhibition, in accordance with an implementation of the present subject matter.

[0011] Figure 3 depicts the effect of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, alone, and in combination at varying weight ratios, on TNF-a inhibition, in accordance with an implementation of the present subject matter.
[0012] Figure 4 depicts the effect of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, alone, and in combination at varying weight ratios, on reactive oxygen species (ROS) inhibition, in accordance with an implementation of the present subject matter.
DETAILED DESCRIPTION OF THE INVENTION
[0013] 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
[0014] 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.
[0015] 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.
[0016] 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".
[0017] 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.

[0018] The term "including" is used to mean "including but not limited to",
"Including" and "including but not limited to" are used interchangeably.
[0019] 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. For example, a weight
ratio of 3:1 to 12:1 is construed to cover not only the explicitly recited ratios of 3:1
and 12:1, but also covers the ranges such as 4:1 to 11:1, 5:1 to 10:1, and ratios such
as 4.5:1, 6.75:1 and all other possible ratios and ranges.
[0020] The term "at least one" is used to mean one or more and thus includes
individual components as well as mixtures/combinations.
[0021] The term "effective dose" refers to the concentration of actives in
combinations which can promote the probiotic growth and inhibit the pathogenic
growth. The term "prebiotic" refers to a a non-digestible food ingredient that
promotes the growth of beneficial microorganisms in the intestines.
[0022] The terms "fibre", "fats", "protein", "vitamin", "mineral", "multigrain" refer
to well-known terms in the field of invention. The term "multigrain" is used to
indicate the presence of more than one type of grain in one product.
[0023] 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.
[0024] 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.

[0025] Conventionally, improvement in gut health is achieved with probiotic foods, such as yogurt, kefir, etc., or through the use of probiotic supplements. However, in a typical modern-day diet, where the food is highly processed, and rich in sugar, managing a healthy, fermented diet is a significant challenge. Although probiotic supplements are being increasingly used, the beneficial microorganism can be killed during processing of the supplements. Prebiotics are being increasingly used as an alternative to overcome the drawbacks associated with the probiotics. Conventionally used prebiotic foods are rich in fibre; when consumed show side effects like bloating or can cause discomfort in the stomach. In fact, it can be even contra productive for people suffering from inflammatory bowel conditions. In addition, the effective dosage of many prebiotics requires several grams, which are not easy to consume or not palatable, thereby leading to a low consumer compliance. Therefore, there exists a need to develop prebiotic compositions, which overcome the drawbacks of the prior art. Therefore, the object of the present disclosure is to provide a therapeutically effective composition, which at low dosages, is effective in improving the gut health of an individual, without imparting side effects, like bloating or gastrointestinal discomfort. Accordingly, the present disclosure provides for compositions comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, which when used in defined w/w ratios is effective in enhancing the gastrointestinal microflora, and consequently enhance health and the gastrointestinal well- being, in addition to imparting anti-oxidant properties, and anti-microbial property.
[0026] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1. In another embodiment of the present disclosure, the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 5:1 to 11:1. In one other embodiment of the present disclosure, the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 7:1 to 11:1. In yet another embodiment of the present

disclosure, the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is 10:1.
[0027] In an embodiment of the present disclosure there is provided a composition as described herein, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 8:1 to 11:1.
[0028] In an embodiment of the present disclosure there is provided a composition as described herein, wherein the Cinnamomum zeylanicum extract has a weight percentage in a range of 0.001-0.1% with respect to the composition. In another embodiment of the present disclosure, the Cinnamomum zeylanicum extract has a weight percentage in a range of 0.01-0.1% with respect to the composition. In yet another embodiment of the present disclosure, the Cinnamomum zeylanicum extract has a weight percentage in a range of 0.02-0.08% with respect to the composition. [0029] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1; and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition.
[0030] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 8:1 to 11:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition.
[0031] In an embodiment of the present disclosure there is provided a composition as described herein, wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition. In another embodiment of the present disclosure, wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.01-0.1% with respect to the composition. In yet another embodiment of the present disclosure, the Glycyrrhiza glabra extract has a weight percentage in a range of 0.03-0.06% with respect to the composition.

[0032] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0033] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 8:1 to 11:1, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0034] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition.
[0035] In an embodiment of the present disclosure there is provided a composition as described herein, wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract. In another embodiment of the present disclosure, the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-11% with respect to the Cinnamomum zeylanicum extract.
[0036] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract.
[0037] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract,

wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract.
[0038] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract. In another embodiment of the present disclosure, the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-22% with respect to the Glycyrrhiza glabra extract. [0039] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract.
[0040] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract.
[0041] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, and wherein the

Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract.
[0042] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 8:1 to 11:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract.
[0043] In an embodiment of the present disclosure there is provided a composition as described herein, wherein the composition further comprises at least one excipient selected from a group consisting of fibre, fats, protein, vitamin, mineral, multigrain flour, and combinations thereof.
[0044] In an embodiment of the present disclosure there is provided a composition comprising: a) Cinnamomum zeylanicum extract; b) Glycyrrhiza glabra extract, and c) at least one excipient selected from a group consisting of fibre, fats, protein, vitamin, mineral, multigrain flour, and combinations thereof, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1.
[0045] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the

Glycyrrhiza glabra extract; and iv) contacting the Cinnamomum zeylanicum extract, and the Glycyrrhiza glabra extract, to obtain the composition. [0046] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; and iv) contacting the Cinnamomum zeylanicum extract, and the Glycyrrhiza glabra extract, to obtain the composition. [0047] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; and iv) contacting the Cinnamomum zeylanicum extract, and the Glycyrrhiza glabra extract, to obtain the composition.
[0048] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; and b) Glycyrrhiza glabra extract, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range

of 10-12% with respect to the Cinnamomum zeylanicum extract, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract, said process comprising: i) obtaining the the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; and iv) contacting the Cinnamomum zeylanicum extract, and the Glycyrrhiza glabra extract, to obtain the composition.
[0049] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; b) Glycyrrhiza glabra extract; and c) at least one excipient selected from a group consisting of fibre, fats, protein, vitamin, mineral, multigrain flour, and combinations thereof, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; iii) obtaining the at least one excipient; and iv) contacting the Cinnamomum zeylanicum extract, the Glycyrrhiza glabra extract, and the at least one excipient, to obtain the composition.
[0050] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; b) Glycyrrhiza glabra extract; and c) at least one excipient selected from a group consisting of fibre, fats, protein, vitamin, mineral, multigrain flour, and combinations thereof, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; iii) obtaining the at least one excipient; and iv) contacting the Cinnamomum zeylanicum extract, the Glycyrrhiza glabra extract, and the at least one excipient, to obtain the composition.
[0051] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; b)

Glycyrrhiza glabra extract; and c) at least one excipient selected from a group consisting of fibre, fats, protein, vitamin, mineral, multigrain flour, and combinations thereof, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; iii) obtaining the at least one excipient; and iv) contacting the Cinnamomum zeylanicum extract, the Glycyrrhiza glabra extract, and the at least one excipient, to obtain the composition. [0052] In an embodiment of the present disclosure there is provided a process for preparing a composition comprising: a) Cinnamomum zeylanicum extract; b) Glycyrrhiza glabra extract; and c) at least one excipient selected from a group consisting of fibre, fats, protein, vitamin, mineral, multigrain flour, and combinations thereof, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1, and wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition, and wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition, and wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract, and wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract, said process comprising: i) obtaining the Cinnamomum zeylanicum extract; ii) obtaining the Glycyrrhiza glabra extract; iii) obtaining the at least one excipient; and iv) contacting the Cinnamomum zeylanicum extract, the Glycyrrhiza glabra extract, and the at least one excipient, to obtain the composition. [0053] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the composition inhibits nitric oxide production, and tumor necrosis factor (TNF)-a production.

[0054] In an embodiment of the present disclosure, the composition as described
herein in the form of a powder.
[0055] In an embodiment of the present disclosure, the composition as described
herein in the form of biscuit.
[0056] In an embodiment of the present disclosure, the composition as described
herein in the form of supplement formulations.
[0057] In an embodiment of the present disclosure, the composition as described
herein, is to improve the gastrointestinal microflora, and correspondingly the gut
health, of an individual.
[0058] In an embodiment of the present disclosure, the composition as described
herein, is to impart an anti-oxidant property.
[0059] In an embodiment of the present disclosure, the composition as described
herein, is to impart an anti-microbial property.
[0060] Although the subject matter has been described with reference to specific
embodiments, this description is not meant to be construed in a limiting sense.
Various modifications of the disclosed embodiments, as well as alternate
embodiments of the subject matter, will become apparent to persons skilled in the
art upon reference to the description of the subject matter. It is therefore
contemplated that such modifications can be made without departing from the spirit
or scope of the present subject matter as defined.
EXAMPLES
[0061] 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.

[0062] The working examples described herein clearly describe the effect of the composition comprising Cinnamomum zeylanicum extract, Glycyrrhiza glabra extract, which when used in defined w/w ratios synergistically enhances the gut health by promoting the growth of probiotic bacteria, and by inhibiting the growth of pathogenic bacteria. The compositions of the present disclosure impart protection against gastrointestinal stresses like oxidative stress, pathogenic stress, and the like.
Example 1: Preparation of microbes
[0063] Individual bacteria; probiotic (Bacillus subtilis CNCM I-2745) and pathogen (E. coli MTCC448) were propagated by culturing in MRS broth (10g/L peptone, 10g/L beef extract,5g/L yeast extract, 20g/L glucose, 2g/L ammonium citrate, 5g/L sodium acetate, 0.1g/L magnesium sulphate, 0.05g/L manganese sulphate and 2g/L dipotassium phosphate) at 37°C for 24hrs.
Example 2: Evaluating the prebiotic activity of the actives, alone and in combination.
[0064] Preparation of the inoculum: Prebiotic potential of selected substances in in-vitro studies were studied by evaluating the effect of herbal extracts (Cinnamomum zeylanicum extract or cinnamon extract; and Glycyrrhiza glabra or licorice extract), alone and in combination, on growth of probiotic bacteria (Bacillus subtilis CNCM I-2745); and the inhibitory effects on pathogen (E. coli MTCC448). For this purpose, inoculation media was prepared without glucose (peptone, 10g/L; beef extract, 8g/L; yeast extract, 4g/L; dipotassium hydrogen phosphate, 2g/L; sodium acetate, 5g/L; tri ammonium citrate, 2g/L; magnesium sulphate, 0.2g/L; manganese sulphate, 0.05g/L) was prepared to which 1000µl of different herbal extracts, as described above, was added to serve as a carbohydrate source. Inoculum media with glucose as a carbohydrate source served as positive control; whereas the media without glucose served as a negative control. The media thus prepared was incubated with test bacteria (Bacillus subtilis CNCM I-2745, and E. coli MTCC448) at a concentration 106 cfu/ml and incubated at 37°C for 16hrs. The optical density (O.D) was measured at 600nm.
[0065] Example 3: Effect of the actives alone, and in combination on LPS stimulated oxidative stress

[0066] RAW 264.7 cells were seeded at 15000cells/well density in 96well plate and incubated at 37°C, 5%CO2. After 80% confluency, cells were treated with combination of selected plant extracts under LPS stimulation for 24hr. Post incubation, cell supernatants were analysed for nitric oxide (NO) production. After incubation, supernatants were collected and used for the estimation of nitric oxide (nitrate and nitrite).
[0067] Preparation of nitrite standard: Prepared the 10mM sodium nitrite solution with distilled water. The stock solution of sodium nitrite (10mM) was diluted with distilled water to prepare a gradient of nitrite standard (0, 1.56, 3.125, 6.25, 12.5, 25, 50 and 100µM).
[0068] Preparation of nitrate standard: Prepared the 20mM sodium nitrate solution with distilled water. The stock solution of sodium nitrite (20mM) was diluted with distilled water to prepare a gradient of nitrite standard (0, 3.125, 6.25, 12.5, 25, 50, 100 and 200µM).
[0069] Assay procedure for total nitric oxide estimation: Standard solution (as described above) and supernatants of different treatments of 50 µl was added to 96 well assay plate containing 50 µl of griess reagent and incubated at room temperature for 10mins. After incubation, the absorbance was measured at 540nm to determine the nitrite concentration in the supernatant as well as the standard. For total nitric oxide estimation, 5 µl of 2%VCl3 (sigma) was added to each well and incubated at 60ºC for 1hr. After incubation, the absorbance was measured at 540nm along with nitrate standard. Total nitric oxide in supernatants were calculated using standard curve.
[0070] Example 4: Effect of the actives alone, and in combination on LPS
mediated pro-inflammation (TNFfn&
[0071] After incubation with actives, supernatants were analysed for the LPS
stimulated secreted cytokine production (TNF- }) by ELISA from R&D systems
(Duo set ELISA catalogue no: TNF-a DY410).
[0072] Preparation of assay plates for TNF- }: Capture antibody of TNF- },
0.2µg/well were used for the coating of 96-wellmicroplate and incubated for
overnight at room temperature. After incubation, aspirated the capture antibody and

washed with wash buffer containing 0.05% tween 20 in PBS (pH-7.2) for 4 times. Then blocked the plate with 300µl of reagent diluent containing 1%BSA in PBS (pH-7.2) for 1hr at room temperature. After incubation, aspirated the reagent diluent and washed with wash buffer for 4 times. The plate was then used for estimation of TNF-}.
[0073] Preparation of TNF- } standards: Recombinant TNF- } were reconstituted with distilled water and allowed for 15min and final concentration was 120ng/ml. Recombinant TNF-a (120ng/ml) was diluted with reagent diluent to prepare a gradient of TNF- } concentrations (0, 9.375, 18.75, 37.5, 75, 150, 300 and 600 pg/ml).
[0074] Assay procedure for estimation of TNF- } production in supernatants: Standard solution (as described above) and supernatants of different actives treatments of 100 µl was added to 96 well containing capture antibody TNF- } and incubated at room temperature for 2hrs. After incubation, the solution was aspirated, and plate was washed with wash buffer for 4 times. Then added the detection antibody of TNF- } (5ng/well) to 96 well plate of TNF- } and incubated at room temperature for 2hrs. After incubation, the solution was aspirated, and plate was washed with wash buffer for 4 times. Added the 100µl of 1:200 diluted streptavidin-HRP (horse radish peroxidase) to 96 well plate and incubated at room temperature for 20 min in dark. The washing step was repeated and 100µl of substrate solution containing hydrogen peroxide and tetramethylbenzidine in 1:1 ratio was added to each well and incubated for 20min at dark. After incubation, added 50µl of 2N sulfuric acid to stop the reaction. Then the calibration standards and samples were measured at 450nm. A calibration curve was recorded with the OD values against the respective cytokines TNF- } concentration. Cytokine production in supernatants were calculated using standard curve.
Example 5: Evaluation of anti-oxidative potential of the actives, alone and in
combination
[0075] RAW 264.7 cells were seeded at 15000 cells/well density in 96-well plate
and incubated at 37°C, 5% CO2. After 80% confluency, cells were treated with
10µg/ml of different herbal extracts (cinnamon extract, and licorice extract, alone

and in combination), and the plates were incubated for for 24h. Quercetin served as a positive control. After incubation, the cells are washed with 1X phosphate buffered saline (PBS) with no Ca, Mg or Hanks balanced salt solution (HBSS) without Ca, and Mg. The plates were then incubated with 25uM 2',7'- dichlorofluorescin diacetate (DCFH-DA) in serum free DMEM for 30min. Post incubation, DCFH-DA was removed, and the cells were washed with PBS/HBSS and incubated in PBS with 100µM H2O2 for 30min. The fluorescence is measured at 480/530nm.The fluorescence intensity (arbitrary unit) was calculated by:
[(Ft30-s-Ft30-c)/Ft30-c] * 100
Ft30-s = fluorescence at time 30 (min) after the addition of hydrogen peroxide to the well treated with herbal extracts; Ft30-c= fluorescence of cells added after the addition of hydrogen peroxide without treatment.
Results and Discussion
[0076] Figure 1 depicts the effect of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, alone, and in combination, at a weight ratio of 10:1 (Cinnamomum zeylanicum extract to Glycyrrhiza glabra extract) on the probiotic bacteria (Bacillus subtilis CNCM I-2745) and pathogen (E. coli MTCC448). From the Figure 1, it can be observed that the combination of cinnamon extract; and licorice extract at weight ratio of 10:1 (0.05% of cinnamon extract and 0.005% of licorice extract) allowed for maximal growth of probiotic bacteria while facilitating the growth of pathogen to a lesser extent, in comparison to the growth exhibited by any of the actives at individual concentration ranges tested, and control (glucose). Although a combination of cinnamon extract and Foeniculum vulgare (saunf) extract was effective in promoting the growth of probiotic bacteria at a weight ratio of 1:1 (0.05% each of cinnamon extract and saunf extract), it can be observed that the combination equally facilitated the growth of the pathogen. In fact, it may be observed that the growth of the pathogen was higher in comparison to the growth exhibited by any of the actives at individual concentration ranges tested, and control (glucose).

[0077] Figure 2 depicts the effect of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, alone, and in combination at varying weight ratios, on nitric oxide (NO) inhibition. From a reading of the Figure 2, it may be observed that the percentage inhibition of nitric oxide with 0.05% of cinnamon was found to be around 29%, while the inhibition of NO was 8% with 0.005% of licorice extract. However, unexpectedly and surprisingly, when a combination of extracts, i.e., 0.05% of cinnamon extract and 0.005% of licorice extract, was used, the percentage inhibition of nitric oxide was found to be around 40%, that is synergistically higher than the effect observed with individual actives alone.
[0078] Figure 3 depicts the effect of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, alone, and in combination at varying weight ratios, on TNF-a inhibition. From a reading of the Figure 3, it may be observed that the percentage inhibition of TNF-a with 0,05% of cinnamon was found to be around 41%, and 16% with 0.005% of licorice extract. However, unexpectedly and surprisingly, when a combination of extracts, i.e., 0.05% of cinnamon extract and 0.005% of licorice extract, was used, the percentage inhibition of TNF-a was found to be around 65%, that is synergistically higher than the effect observed with individual actives alone.
[0079] Further, similar experiments were repeated at varying w/w ratios, and the results are presented in Figure 4. The percentage inhibition of reactive oxygen species (ROS), exhibited by the individual actives i.e., cinnamon extract (C) and licorice extract (L) alone and in combination, at varying weight ratios is herewith presented below in Table 1, and Figure 4.

[0080] As per Table 1, the combination of two extracts with weight ratios of 10:1 (0.05% of C and 0.005% of L) and 9:1 (0.0495% of C and 0.0055% of L) provide a synergistic effect as compared to the additive effect of the respective constituents at said weight percentage. It can be observed that the combinations of two extracts with weight ratio 0.10:1 (0.005% of C and 0.0495% of L) does not exhibit any synergistic effect. Also, no significant increase in the percentage inhibition of ROS can be observed for the combinations with weight ratios of 1.5:1 and 0.66:1. Therefore, it is evident from Table 1 that a mere combination of Cinnamomum zeylanicum extract, and Glycyrrhiza glabra extract in any weight ratio does not yield synergistic results. On the other hand, a combination having Cinnamomum zeylanicum extract, and Glycyrrhiza glabra extract in a weight ratio in a range of 3:1 to 12:1 results in synergistic inhibition of ROS production, NO production, and TNF-a production. [0081] Overall, from among the actives tested, alone and in combination, it can be observed that a combination of Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract at a weight ratio of 10:1 allowed for maximal growth of probiotic bacteria by inhibiting the growth of pathogen, in comparison to the growth exhibited by any of the actives at individual concentration ranges tested, and control (glucose). The above data substantiates that the proposed combination is effective in enhancing the gastrointestinal microflora, in addition to imparting anti-oxidant properties, and anti-microbial property, and the same has been experimentally established as described herein.
Advantages of the present disclosure
[0082] The present disclosure discloses a composition comprising Cinnamomum zeylanicum extract; and Glycyrrhiza glabra extract, which when used in defined weight ratios is effective in imparting anti-oxidant and anti-microbial properties, in addition to improving the gut health of the individual by promoting probiotic

bacterial growth by inhibiting the growth of pathogen. Further, unlike the conventionally used prebiotics which are mostly made of fibre, and are hence unpalatable, the composition of the present disclosure can be used in a variety of food products such as confectionery, bakery product, dairy product, meal replacement product, soup product, dehydrated and culinary food, frozen food, canned food, snack food, dips, oils and fats, and seasoning. Further, the composition of the present disclosure does not use any synthetics to improve the gut health, thereby overcoming the problems associated with synthetics, such as toxicity. The present disclosure also provides for a convenient process for preparation of the composition.

I/We Claim:
1. A composition comprising:
a) Cinnamomum zeylanicum extract; and
b) Glycyrrhiza glabra extract,
wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 3:1 to 12:1.
2. The composition as claimed in claim 1, wherein the Cinnamomum zeylanicum extract to the Glycyrrhiza glabra extract weight ratio is in a range of 8:1 to 11:1.
3. The composition as claimed in claim 1, wherein the Cinnamomum zeylanicum extract has a weight percentage is in a range of 0.001-0.1% with respect to the composition.
4. The composition as claimed in claim 1, wherein the Glycyrrhiza glabra extract has a weight percentage in a range of 0.001-0.1% with respect to the composition.
5. The composition as claimed in claim 1, wherein the Cinnamomum zeylanicum extract comprises polyphenol in a range of 10-12% with respect to the Cinnamomum zeylanicum extract.
6. The composition as claimed in claim 1, wherein the Glycyrrhiza glabra extract comprises glycyrrhizic acid in a range of 20-23% with respect to the Glycyrrhiza glabra extract.
7. The composition as claimed in claim 1, wherein the composition further comprises at least one excipient selected from a group consisting of fibre, fat, protein, flavour, vitamin, mineral, multigrain flour, and combinations thereof.
8. A process for preparing the composition as claimed in claim 1, said process comprises:

a) obtaining the Cinnamomum zeylanicum extract;
b) obtaining the Glycyrrhiza glabra extract; and
c) contacting the Cinnamomum zeylanicum extract, and the Glycyrrhiza glabra extract, to obtain the composition.

9. A process for preparing the composition as claimed in claim 7, said process
comprises:
a) obtaining the Cinnamomum zeylanicum extract;
b) obtaining the Glycyrrhiza glabra extract;
c) obtaining the at least one excipient; and
d) contacting the Cinnamomum zeylanicum extract, the Glycyrrhiza glabra extract, and the at least one excipient, to obtain the composition.
10. The composition as claimed in any one of the claims 1-7, wherein the
composition inhibits nitric oxide production, and Tumor necrosis factor-}
(TNF-}) production.