FIELD OF INVENTION
[001] The present disclosure relates to the composition for disease management in general and composition for inhibiting oxidative stress in particular. There is provided a composition comprising pomegranate extract and at least one flavone for oxidative stress management.
BACKGROUND OF THE INVENTION
[002] Free radicals, also known as reactive oxygen species, are common by-products
of various cellular processes in the body that involves the use of oxygen. These
radicals are highly reactive due to the presence of unpaired electron in their outer
orbits. Apart from the natural biological processes, exposure to environmental stress
such as UV and heat also lead to substantial increase of free radical concentration in
body. Some of the common examples include, peroxide, superoxide, hydroxyl
radicals, and singlet oxygen. Under normal conditions, the inbuilt antioxidant
mechanism of the body manages the free radicals by neutralizing them and thereby
prevents any damage.
[003] Oxidative stress is a result of imbalance between production of free radical and
ability of the body to neutralize these radicals. It can eventually lead to manifestation
of many chronic diseases such as atherosclerosis, cancer, diabetes, rheumatoid
arthritis, post-ischemic perfusion injury, myocardial infarction, cardiovascular
diseases, chronic inflammation, stroke and septic shock, and aging in humans (Uttara
et. al., Current Neuropharmacology. 2009, 7(1): 65-74).
[004] Antioxidants are molecules that neutralize the free radicals and in turn prevent
the oxidation of other molecules. Apart from being naturally present in fruits and
vegetables, antioxidants are also available externally as dietary supplements, such as
beta-carotene, lutein, lycopene, vitamin A, and vitamin C.
[005] WO2007004229 reveals a composition comprising pomegranate and its
constituents for promotion of skin repair.

[006] JPH05320037 discloses the use of pomegranate bark in preparation of anti-oxidant agent.
[007] The current synthetic dietary supplements that are available in the market are associated with innumerable health related risks, including liver damage, birth defects, nausea, nerve damage, so on and so forth. Due to the reasons indicated above, use of plants and its extracts for preparation of antioxidant formulation, is highly desirable and under extensive exploration.
SUMMARY OF INVENTION
[008] In an aspect of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[009] In an aspect of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; and (c) contacting the pomegranate extract and the at least one flavone to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5. [0010] 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
[0011] The following drawings form a 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.
[0012] Figure 1 depicts a graph that shows cytotoxic effects of different
concentrations of plant extracts on RAW 264.7 cells, in accordance with an
embodiment of the present disclosure.
[0013] Figure 2 depicts a graph that shows the effect of plant extracts on the
production of ROS in H2O2 induced RAW 264.7 cells, in accordance with an
embodiment of the present disclosure.
[0014] Figure 3 depicts a graph that shows the effect of luteolin and pomegranate
extracts on inhibition of ROS in H2O2 induced RAW 264.7 cells, in accordance with
an embodiment of the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0015] 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
[0016] 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.
[0017] 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.

[0018] 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”.
[0019] 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.
[0020] The term “including” is used to mean “including but not limited to”.
“Including” and “including but not limited to” are used interchangeably.
[0021] Carriers are substances that serve as mechanisms to improve the delivery and
the effectiveness of the drug.
[0022] A diluent (also referred to as filler, dilutant, or thinner) is a diluting agent.
[0023] An excipient is an inactive substance that serves as a vehicle or medium for a
drug or other active substance. Excipients include colouring agents, humectants,
preservatives, emollients and combinations thereof.
[0024] ROS or reactive oxygen species are free radicals that are generated in body as
a natural by-product of the normal metabolism of oxygen. They play important role in
cell signalling and homeostasis. Increased concentration of ROS in body due to
various external and internal stress result in oxidative stress.
[0025] RAW 264.7 is an adherent murine macrophage cell line. They mimic the
properties of human macrophages and therefore, are used in various cell based
experimentations.
[0026] 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.

[0027] Reactive oxygen species (ROS) have a dual role in being deleterious as well
as beneficial for heath. Most of the metabolic reactions result in release of ROS,
which in turn have important role in cell signalling and homeostasis. However, an
abnormal increase in the levels of ROS due to different internal and external factors
can lead to various health implications. These free radicals can react with fundamental
molecules such as lipids, proteins and DNA and give way to complicated chronic
disorders. Diseases such as diabetes, Alzheimer’s and atherosclerosis have been
actively associated with oxidative stress.
[0028] Luteolin is a flavone that belong to the class of flavonoid and are mostly
present in leaves. Celery, broccoli, parsley, and carrots are few common sources of
luteolin. It has multiple beneficiary effects including anti-inflammation, anti-allergy,
and anti-cancer. Plants rich in luteolin have been used in Chinese traditional medicine
for treating various diseases such as hypertension, inflammatory disorders, and cancer
(Lin Y et.al., Current Cancer Drug Targets. 2008 8(7): 634-436). In addition, the
flavone molecule also possesses anti-oxidative potential.
[0029] The current specification discloses a synergistic composition comprising
luteolin and pomegranate extract which has been identified as a potent antioxidant. the
composition combats the effect of oxidative stress by lowering the levels of free
radicals.
[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
comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least
one flavone to the pomegranate extract w/w ratio in said composition is in the range of
1:0.5- 1:5.
[0032] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least

one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0033] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5- 1:5.
[0034] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:1. [0035] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:2.5. [0036] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:4. [0037] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5. [0038] In an embodiment of the present disclosure, the pomegranate extract, an essential component of the composition described herein, contains punicalagin having weight percentage in the range of 2- 10%.
[0039] In an embodiment of the present disclosure, the pomegranate extract, an essential component of the composition described herein, contains punicalagin having weight percentage in the range of 8- 15%.
[0040] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003-0.00063%, weight percentage of the pomegranate extract in said composition is in the

range of 0.00016-0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0041] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003-0.00063%, weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0042] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003-0.00063%, weight percentage of the pomegranate extract in said composition is in the range of 0.0005- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0043] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003-0.00063%, weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5- 1:5.
[0044] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003-0.00063%, weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5.
[0045] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003-

0.00063%, weight percentage of the pomegranate extract in said composition is in the
range of 0.0005- 0.0032%, and the at least one flavone to the pomegranate extract
w/w ratio in said composition is 1:5.
[0046] In an embodiment of the present disclosure, the at least one flavone, an
essential component of the composition described herein, is selected from a group
consisting of luteolin, nobiletin, tangeretin, and combinations thereof.
[0047] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0048] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0049] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is 1:1.
[0050] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is 1:2.5.
[0051] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is 1:4.
[0052] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is 1:5.
[0053] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein weight percentage of
luteolin in said composition is in the range of 0.0003- 0.00063%, weight percentage of
the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and

luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0054] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) luteolin, wherein weight percentage of luteolin in said composition is in the range of 0.0003- 0.00063%, weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0055] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5. [0056] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5. [0057] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5- 1:5. [0058] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:1. [0059] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:2.5.

[0060] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:4. [0061] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5. [0062] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0063] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0064] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.0005- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5-1:5.

[0065] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5- 1:5.
[0066] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5.
[0067] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0068] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0069] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is 1:1.
[0070] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least

one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is 1:5.
[0071] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of luteolin in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5-1:5.
[0072] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of luteolin in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0073] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of luteolin in said composition is in the range of 0.0003- 0.00063%, the weight percentage of the pomegranate extract in said composition is in the range of 0.0005- 0.0032%, and luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0074] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; and (c) contacting the pomegranate extract and the at least one flavone to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.

[0075] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; and (c) contacting the pomegranate extract and the at least one flavone to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5-1:5. [0076] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; and (c) contacting the pomegranate extract and the at least one flavone to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5- 1:5. [0077] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; and (c) contacting the pomegranate extract and the at least one flavone to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5.
[0078] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) luteolin, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining luteolin; and (c) contacting the pomegranate extract and luteolin to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0079] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) luteolin, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining luteolin; and (c) contacting the pomegranate extract and luteolin to obtain said composition,

wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0080] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) luteolin, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining luteolin; and (c) contacting the pomegranate extract and luteolin to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is 1:5. [0081] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) at least one flavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, the at least one flavone, the at least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5. [0082] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) at least one flavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, the at least one flavone, the at least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5. [0083] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) at least one flavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least

one flavone; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, the at least one flavone, the at least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5- 1:5. [0084] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) at least one flavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining the at least one flavone; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, the at least one flavone, the at least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5.
[0085] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) luteolin; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining luteolin; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, luteolin, the at least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0086] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) luteolin; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining luteolin; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, luteolin, the at

least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0087] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; (ii) luteolin; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the pomegranate extract; (b) obtaining luteolin; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the pomegranate extract, luteolin, the at least one carrier, the at least one diluent and the at least one excipient to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is 1:5.
[0088] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein the at least one flavone to the at least pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0089] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein the at least one flavone to the at least pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0090] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate

extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein the at least one flavone to the at least pomegranate extract w/w ratio in said composition is 1:5. [0091] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein weight percentage of the at least one flavone in said composition is in the range of 0.0003- 0.00063%, weight percentage of the pomegranate extract in said composition is in the range of 0.00016- 0.0032%, and the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0092] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) luteolin, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of luteolin; and (c) contacting solution A and solution B to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5.
[0093] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) luteolin, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of luteolin; and (c) contacting solution A and solution B to obtain said composition, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5.
[0094] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting

solution A and solution B to obtain said composition, wherein said solution A is prepared in an organic solvent.
[0095] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution A is prepared in an inorganic solvent.
[0096] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution B is prepared in an organic solvent.
[0097] In an embodiment of the present disclosure, there is provided a method for preparing a composition comprising: (i) pomegranate extract; and (ii) at least one flavone, said method comprising: (a) obtaining a solution A of the pomegranate extract; (b) obtaining a solution B of the at least one flavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution B is prepared in an inorganic solvent.
[0098] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said composition inhibits free radical production. [0099] In an embodiment of the present disclosure, there is provided a composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5, and said composition inhibits free radical production.

[00100] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least
one flavone to the pomegranate extract w/w ratio in said composition is in the range of
1:2.5- 1:5, and said composition inhibits free radical production.
[00101] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least
one flavone to the pomegranate extract w/w ratio in said composition is 1:5, and said
composition inhibits free radical production.
[00102] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and
said composition inhibits free radical production.
[00103] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the
pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5, and
said composition inhibits free radical production.
[00104] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; and (b) luteolin, luteolin to the pomegranate
extract w/w ratio in said composition is 1:5, and said composition inhibits free radical
production.
[00105] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier;
(d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone
to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5,
and said composition inhibits free radical production.
[00106] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier;
(d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone

to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5,
and said composition inhibits free radical production.
[00107] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier;
(d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone
to the pomegranate extract w/w ratio in said composition is 1:5, and said composition
inhibits free radical production.
[00108] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least
one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract
w/w ratio in said composition is in the range of 1:0.5- 1:5, and said composition
inhibits free radical production.
[00109] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least
one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract
w/w ratio in said composition is in the range of 1:1.5- 1:5, and said composition
inhibits free radical production.
[00110] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least
one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract
w/w ratio in said composition is 1:5, and said composition inhibits free radical
production.
[00111] In an embodiment of the present disclosure, there is provided a composition
for use in preparing formulations, said composition comprising: (a) pomegranate
extract; and (b) at least one flavone, wherein the the at least one flavone to the
pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and
said formulations inhibit oxidative stress.
[00112] In an embodiment of the present disclosure, there is provided a composition
for use in preparing formulations, said composition comprising: (a) pomegranate

extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5, and said formulations inhibit oxidative stress.
[00113] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:2.5-1:5, and said formulations inhibit oxidative stress.
[00114] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; and (b) at least one flavone, wherein the the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5, and said formulations inhibit oxidative stress.
[00115] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract having weight percentage in said composition in the range of 0.00016-0.0032%; and (b) at least one flavone having weight percentage in said composition in the range of 0.0003- 0.00063%, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress.
[00116] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress.
[00117] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the pomegranate extract w/w ratio in said

composition is in the range of 1:1.5- 1:5, and said formulations inhibit oxidative stress.
[00118] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; and (b) luteolin, wherein luteolin to the pomegranate extract w/w ratio in said composition is 1:5, and said formulations inhibit oxidative stress. [00119] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract having weight percentage in said composition in the range of 0.00016-0.0032%; and (b) luteolin having weight percentage in said composition in the range of 0.0003-0.00063%, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress.
[00120] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress.
[00121] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5, and said formulations inhibit oxidative stress.
[00122] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w

ratio in said composition is in the range of 1:2.5- 1:5, and said formulations inhibit oxidative stress.
[00123] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) at least one flavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is 1:5, and said formulations inhibit oxidative stress. [00124] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract having weight percentage in the range of 0.00016- 0.0032%; (b) at least one flavone having weight percentage in the range of 0.0003- 0.00063%; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the at least one flavone to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress.
[00125] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress. [00126] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:1.5- 1:5, and said formulations inhibit oxidative stress. [00127] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract; (b) luteolin; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is 1:5, and said formulations inhibit oxidative stress.

[00128] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations, said composition comprising: (a) pomegranate extract having weight percentage in the range of 0.00016- 0.0032%; (b) luteolin having weight percentage in the range of 0.0003- 0.00063%; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein luteolin to the pomegranate extract w/w ratio in said composition is in the range of 1:0.5- 1:5, and said formulations inhibit oxidative stress.
[00129] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be used in preparation of formulations for topical application.
[00130] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be used in preparation of formulations, and said formulations can be used as dietary supplements. [00131] 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
[00132] 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
Material and Methods
[00133] Plant extracts: Ashwagandha, noni extract, cooca oleoresin, Bacopa monnieri 30%, goutickola 20%, Rose hip extract, and cocoa powder were procured from Bioveda Naturals Pvt. Ltd. Moringa leaf extract was procured from Ramona Cosmaceuticals P. Ltd. Apigenin 98% and luteolin 98% were procured from Sigma. Pomegranate extract was procured from Natural Remedies Pvt Ltd. and comprises approximately 10% (w/w) of punicalagin and 2.5% (w/w) of ellagic acid. [00134] Cell viability assay: RAW 264.7 (ATCC), a murine leukemic cell line, was used for all the experiments. The cells were seeded onto a 96 well plate at a density of 15000 cells /well in complete DMEM (Dulbecco's Modified Eagle Medium from Invitrogen) supplemented with 10% fetal bovine serum (FBS) and the plate was incubated at 37 °C/ 5% CO2. Upon reaching the confluency of 80%, the cells were treated with different plant extracts, viz. ashwagandha, noni extract, cocoa oleoresin, Bacopa monnieri, goutickola, rose hip extract, moringa leaf extract, apegenin, luteolin, cocoa powder, and pomegranate extract. Each of the actives were tested at concentrations of 1 mg/ml, 0.5 mg/ml, 0.2 mg/ml, 0.05 mg/ml, 0.01 mg/ml, 0.05 mg/ml, 0.001 mg/ml in treatment media (Dulbecco's Modified Eagle Medium supplemented with 1% fetal bovine serum) and the plate was incubated at 37 °C/ 5% CO2 for a time period of 24 hours. Next day the cells were washed with calcium and magnesium free phosphate buffer solution with pH 7.0. After washing, the cell viability was determined by addition of CCK-8 reagent procured from Dojindo molecular technologies, Inc. 1:10 dilution of the reagent was prepared in serum free DMEM, and 100 ul of the diluted reagent was added to the wells. The plate was

incubated at 37 °(75% CO2 for 1 hour. Post incubation, absorbance was measured at 450 nm using a microplate reader.
[00135] The result of the cell viability assay has been depicted in Figure 1. The values greater than 90% to that of the control cells were considered as safe or non-cytotoxic dose. As can be observed, most of the plant extracts display cytotoxicity at higher concentration. Based on the results obtained, the non-cytotoxic concentration for all the tested actives was determined. Table 1 below represents the safe dose or the maximum dose at which the plant extract exhibits almost 90% cell viability as compared to the vehicle control (VC), which in this case is DMSO.
[00136] Determination of inhibition of ROS production by individual plant extracts: After confirming the non-toxic concentration, the plant extracts were further tested for their ability to inhibit ROS production. Healthy and viable RAW 264.7 cells were plated onto a 96 well plate at a density of 15000 cells/well and incubated at 37 °C, 5%

CO2 till attainment of 80% confluency. The cells were then treated with different plant extracts at their non-toxic concentration (Table 1) for a period of 24 hours. Next day, the cells were washed with calcium and magnesium free PBS or calcium and magnesium free HBSS, followed by incubation with 25uM of 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA from Sigma-Aldrich, prepared in serum free DMEM) for 30 minutes at 37 °(75% CO2. Post incubation, the media containing DCFH-DA was removed and the cells were washed with PBS/HBSS. 100UM hydrogen peroxide (H2O2) prepared in PBS was added to the wells for 30 minutes for inducing oxidative-stress. Finally, the fluorescence was measured at a wavelength of 480/530nm and the fluorescence intensity was measured using the formula below:
[(Ft30-s - Ft30-c)/Ft30-c x 100)]
Ft30-s = fluorescence reading at 30minutes time point of the cells that were treated with plant extracts and stimulated with hydrogen peroxide.
Ft30-c = fluorescence reading at 30-minute time point of the cells not treated with plant extract but stimulated with hydrogen peroxide.
[00137] Figure 2 represents a graph depicting the effect of plant extract on ROS production. Untreated cells stimulated with H2O2 alone serves as positive control for the experiment. Amongst all the tested actives, luteolin and pomegranate extract displayed least ROS production. As compared to untreated cell which shows 100% ROS production, luteolin and pomegranate extract exhibit only 43.08% and 24.88% of ROS production respectively. In other words, luteolin and pomegranate extract display 57% and 75.2% inhibition in production of ROS, respectively as compared to the untreated cells which exhibit 0% inhibition in ROS production.
Example 3
[00138] Effect of combination of luteolin and pomegranate extract on ROS inhibition: Seeding of RAW 264.7 cells was performed in a similar manner as

described in Example 2. Once confluent, the cells were treated with luteolin and pomegranate extract individually and in combination for a period of 24 hours. The concentrations tested are indicated in Table 2. All the steps followed for processing of the cells and measurement of ROS post treatment were same as stated earlier in
[00139] Figure 3 depicts a graph that displays the individual and combined effect of luteolin and pomegranate extract on ROS inhibition. The values of ROS inhibition depicted in Figure 3 has further been illustrated in Table 2 above. As can be observed, pomegranate extract individually is not able to significantly inhibit ROS. The highest inhibition of mere 26.48% is observed at maximum concentration of 0.05 mg/ml of pomegranate extract. As for luteolin, significant inhibition can be observed at

concentration higher than 0.01 mg/ml due to high cytotoxicity of the active at these concentrations. There is a drop in the percentage inhibition of ROS as the concentration of luteolin is decreased from 0.025 mg/ml to 0.005 mg/ml. Therefore, both the plant extracts exhibit poor percentage inhibition of ROS when tested individually at lower concentrations. However, unexpectedly and surprisingly, the combination of luteolin and pomegranate extract synergistically enhances percentage inhibition of ROS, particularly at w/w ratio of 1:5 (0.005 mg/ml of luteolin and 0.025 mg/ml of pomegranate extract) and w/w ratio of 1:1 (0.005 mg/ml each),. [00140] From the above set of experimental data, it is apparent that, luteolin and pomegranate extract when used in combination are able to inhibit ROS even at lower concentrations. This synergism of the actives is highly apparent at w/w ratio of 1:5 of luteolin to pomegranate extract.
Advantages of the present disclosure: Overall, the present disclosure provides a composition comprising a flavone, i.e. luteolin, and pomegranate extract, which at specific w/w ratios is able enhance the percentage inhibition of free radicals. The composition employs lower concentration of both the aforementioned actives and displays unexpected synergistic percentage inhibition of ROS. The composition can further be employed in preparation of formulations that can inhibit free radical production and thereby aid in oxidative stress management. The composition presents an added advantage of safe consumption, as both the actives are of plant origin.

A composition comprising:
(a) pomegranate extract; and
(b) at least one flavone,
wherein the at least one flavone to the pomegranate extract w/w ratio in said
composition is in the range of 1:0.5 –1:5.
The composition as claimed in claim 1, wherein the at least one flavone to the
pomegranate extract w/w ratio in said composition is in the range of 1:1.5 -1:5.
The composition as claimed in claim 1, wherein the at least one flavone to the
pomegranate extract w/w ratio in said composition is in the range of 1:2.5 -1:5.
The composition as claimed in claim 1, wherein the at least one flavone is
selected from a group consisting of luteolin, nobiletin, tangeretin, and
combinations thereof.
The composition as claimed in claim 1, wherein the at least one flavone is
luteolin.
The composition as claimed in claim 1, further comprising:
(a) at least one carrier;
(b) at least one diluent; and
(c) at least one excipient.
A method for preparing a composition as claimed in claim 1, said method comprising:
(a) obtaining pomegranate extract;
(b) obtaining at least one flavone; and
(c) contacting the pomegranate extract and the at least one flavone to obtain said composition.
A method for preparing a composition as claimed in claim 6, said method
comprising:
(a) obtaining pomegranate extract;

(b) obtaining at least one flavone;
(c) obtaining at least one carrier;
(d) obtaining at least one diluent;
(e) obtaining at least one excipient; and
(f) contacting the pomegranate extract, the at least one flavone, the at least one carrier, the at least one diluent, and the at least one excipient to obtain said composition.
The composition as claimed in any of claims 1-6, wherein said composition
inhibits free radical production.
The composition as claimed in any of the claims 1-6, wherein said composition
is used for preparing formulations to inhibit oxidative stress.
The composition as claimed in claim 10, wherein said formulations are for
topical application.
The composition as claimed in claim 10, wherein said formulations are used as
dietary supplements.