FIELD OF THE INVENTION
[001] The present disclosure relates to the field of cosmetics and formulations for skin aging, in particular. There is provided a composition comprising apple fruit cellular extract and at least one peptide and uses thereof.
BACKGROUND OF THE INVENTION
[002] Skin aging is a multifactorial biological process governed by a variety of factors. These factors include both exogenous and endogenous factors. The different endogenous factors affecting skin aging include genetic factors, cellular metabolism, different hormones, and metabolic processes. Exogenous factors include chronic UV exposure (photo aging), pollution, and exposure to chemicals.
[003] At the metabolic level, skin aging is characterized primarily by a decrease in synthesis of extracellular matrix (ECM) proteins like collagen and elastin, increase in protease activity leading to degradation of ECM, and decrease in epidermal turnover. The outermost layer of the skin is the stratum corneum which is composed of dead, keratin-filled cells that have migrated outward from the basal layer. The stratum corneum maintains the homeostasis of water and other materials in the skin. Major components of stratum include lipids, stratum corneum proteins, involucrin, loricrin, enzymes, and water. The property of skin elasticity is dependent on the fraction of water retained in stratum. Disturbance in stratum constituents results in wrinkled and dry skin, which can result from reduced synthesis of involucrin, leading to the aging of the skin.
[004] Several strategies have been adopted to prevent skin aging and further research is going on. One of the strategies primarily used is to maintain collagen levels, either by increasing collagen synthesis or decreasing collagen

degradation. Another strategy that can be used is to target the involucrin protein which maintains the structure of stratum corneum and cell envelope.
SUMMARY OF THE INVENTION
[005] The subject matter disclosed here is directed towards the synergistic composition comprising apple fruit cellular extract and at least one peptide. [006] In an aspect of the present disclosure, there is provided a composition to inhibit skin-aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[007] In an aspect of the present disclosure, there is provided a method of preparing a composition to inhibit skin-aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 -1:80.
[008] In an aspect of the present disclosure, there is provided a method of preparing a composition to inhibit skin-aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers, diluents, and excipients; and (iv) contacting the at least one peptide, the apple fruit cellular extract, suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80. [009] In an aspect of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition to inhibit skin-aging comprising: (i) at least one peptide; (ii) apple fruit cellular extract;

and (iii) suitable carriers, diluents and excipients; and (b) contacting said composition with skin, wherein the at least one one peptide to the apple fruit stem cell extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[010] 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
[011] 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.
[012] Figure 1 depicts the chromatogram of chlorogenic acid used as a standard, in
accordance with an embodiment of present disclosure.
[013] Figure 2 depicts the HPLC chromatogram of apple callus lyophilized powder,
in accordance with an embodiment of present disclosure.
[014] Figure 3 depicts the effect of peptide, apple fruit cellular extract, and
combinations thereof on involucrin levels in accordance with an embodiment of
present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[015] 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
[016] 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.
[017] 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.
[018] 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”.
[019] 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.
[020] The term “including” is used to mean “including but not limited to”.
“Including” and “including but not limited to” are used interchangeably.
[021] 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.

[022] Cosmetically suitable carriers, diluents, and excipients are selected from the
group consisting of sunscreen agents, antioxidants, stabilizers, emollients, solvents,
preservatives and combinations thereof.
[023] Carriers are substances that serve as mechanisms to improve the delivery and
the effectiveness of drugs.
[024] A diluent (also referred to as filler, dilutant, or thinner) is a diluting agent.
[025] An excipient is an inactive substance that serves as the vehicle or medium for
a drug or other active substance. Excipients include colouring agents, preservatives,
humectants, and stabilizers.
[026] A composition comprising “synergistic activity” or a “synergistic
composition” is a combination of compounds which exhibits increased biological or
functional activity as a non-linear multiple of the biological or functional activity of
the individual compounds. In other words, the combined biological or functional
activity of two or more compounds being tested is significantly greater than the
expected result based on independent effects of the compounds when tested
separately. Synergy may be apparent only at some ranges or concentrations.
[027] 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.
[028] The present disclosure relates to the problem of skin aging which is
influenced by a variety of factors resulting in reduced synthesis of extracellular
matrix (ECM) proteins like collagen and elastin. The present disclosure provides a
synergistic combination of apple fruit cellular extract and at least one peptide which
enhance involucrin levels and thus, resulting in a solution to the problem of skin
aging.
[029] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular

extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in
said composition is in the range of 1:20-1:80.
[030] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular
extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in
said composition is 1:30.
[031] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular
extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in
said composition is 1:50.
[032] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cell cellular
extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in
said composition is 1:75.
[033] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) a peptide having sequence Ile-Glu-Pro-Asp; and (b)
apple fruit cellular extract, wherein the peptide to the apple fruit cellular extract w/w
ratio in said composition is in the range of 1:20-1:80.
[034] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein the
apple fruit cellular extract comprises chlorogenic acid having weight percentage in
the range of 0.0008-0.0015%, and the at least one peptide to the apple fruit cellular
extract w/w ratio in said composition is in the range of 1:20-1:80.
[035] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular
extract, wherein weight percentage of the at least one peptide in said composition is
in the range of 0.00001-0.0001%, and weight percentage of the apple fruit cellular
extract in the said composition is in the range of 0.0005% to 0.007%, and the at least

one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[036] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide in said composition is 0.00002 %, and weight percentage of the apple fruit cellular extract in said composition is in the range of 0.0005% – 0.007%, and wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[037] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide in said composition is 0.00004 %, and weight percentage of the apple fruit cellular extract in said composition is in the range of 0.0005% – 0.007%, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[038] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide in said composition is 0.00008 %, and weight percentage of the apple fruit cellular extract in said composition is in the range of 0.0005% – 0.007%, and peptide to apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[039] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide in said composition is in the range of 0.00001% - 0.0001%, and weight percentage of the apple fruit cellular extract in said composition is 0.0006%, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[040] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein

weight percentage of the at least one peptide in said composition is in the range of 0.00001% - 0.0001%, and weight percentage of the apple fruit cellular extract in said composition is 0.002%, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[041] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide in said composition is in the range of 0.00001% - 0.0001%, and weight percentage of the apple fruit cellular extract in said composition is 0.006%, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[042] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide is 0.00002 %, weight percentage of the apple fruit cellular extract is 0.0006%, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:30.
[043] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide is 0.00004 %, weight percentage of the apple fruit cellular extract is 0.002%, and the at least one peptide to apple fruit cellular extract w/w ratio in said composition is 1:50.
[044] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein weight percentage of the at least one peptide is 0.00008 %, and weight percentage of the apple fruit cellular extract is 0.006%, and peptide to apple fruit cellular extract w/w ratio in said composition is 1:75.
[045] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the at least one peptide

to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[046] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:30.
[047] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:50.
[048] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:75.
[049] In an embodiment of present disclosure, there is provided a composition to inhibit skin aging comprising: (a) a peptide having amino acid sequence Ile-Glu-Pro-Asp; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the peptide to the the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80.
[050] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) a peptide having amino acid sequence Ile-Glu-Pro-Asp; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the apple fruit extract comprises chlorogenic acid having weight percentage in the range of 0.0008 % - 0.0015 %.
[051] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) a peptide having amino acid sequence Ile-Glu-Pro-Asp; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the peptide in said composition is in the

range of 0.00001-0.0001%, and weight percentage of the apple fruit cellular extract in the said composition is in the range of 0.0005% to 0.007%, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20-1:80.
[052] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein the at least one peptide weight percentage in said composition is 0.00002 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005 % - 0.007 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[053] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the at least one peptide in said composition is 0.00004 %, weight percentage of the apple fruit cellular extract in said composition is in the range of 0.0005 % - 0.007 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[054] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the at least one peptide in said composition is 0.00008 %, weight percentage of the apple fruit cellular extract is in the range of 0.0005 % - 0.007 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[055] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the

at least one peptide in said composition is in the range of 0.00001 % - 0.0001 %, weight percentage of the apple fruit cellular extract is 0.0006 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[056] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; and (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the at least one peptide in said composition is in the range of 0.00001 % - 0.0001 %, and weight percentage of the apple fruit cellular extract is 0.002 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[057] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; and (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the at least one peptide in said composition is in the range of 0.00001 % - 0.0001 %, weight percentage of the apple fruit cellular extract is 0.006 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80.
[058] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the at least one peptide in said composition is 0.00002 %, weight percentage of the apple fruit cellular extract is 0.0006 %, and the at least one peptide to the apple fruit cellular extract w/w in said composition is 1:30.
[059] In an embodiment of the present disclosure, there is provided a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage of the at least one peptide in said composition is 0.00004 %, weight percentage of the apple

fruit cellular extract is 0.002 %, and the at least one peptide to the apple fruit cellular
extract w/w ratio in said composition is 1:50.
[060] In an embodiment of the present disclosure, there is provided a composition to
inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract;
and (c) suitable carriers; (d) diluents; and (e) excipients, wherein weight percentage
of the at least one peptide in said composition is 0.00008 %, weight percentage of the
apple fruit cellular extract is 0.006 %, and the at least one peptide to the apple fruit
cellular extract w/w ratio in said composition is 1:75.
[061] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be for topical application.
[062] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in the form of an aerosol.
[063] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in the form of a lotion.
[064] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in form of an oil.
[065] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in the form of a serum.
[066] In an embodiment of the present disclosure, there is provided a method of
preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and
(b) apple fruit cellular extract, said method comprising: (i) obtaining at least one
peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one
peptide and apple fruit cellular extract to obtain said composition, wherein w/w ratio
of the at least one peptide to the apple fruit cellular extract in said composition is in
the range of 1:20 – 1:80.
[067] In an embodiment of the present disclosure, there is provided a method of
preparing a composition for inhibiting skin aging comprising: (a) at least one peptide;
and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one

peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:30.
[068] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:50.
[069] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:75. [070] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) a peptide having amino acid sequence Ile-Glu-Pro-Asp; and (b) apple fruit cellular extract, said method comprising: (i) obtaining a peptide having amino acid sequence Ile-Glu-Pro-Asp; (ii) obtaining apple fruit cellular extract; and (iii) contacting the peptide and apple fruit cellular extract to obtain said composition, wherein w/w ratio of peptide to apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[071] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one

peptide and apple fruit cellular extract to obtain said composition, wherein the apple fruit cellular extract comprises chlorogenic acid having weight percentage in the said composition in the range of 0.0008 % - 0.0015 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[072] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%- 0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[073] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00002 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%-0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[074] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least

one peptide weight percentage in said composition is 0.00004 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%-0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[075] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00008 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%-0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[076] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%-0.0001%, the apple fruit cellular extract weight percentage in said composition is 0.0006 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[077] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, the

apple fruit cellular extract weight percentage in said composition is 0.002 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[078] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, the apple fruit cellular extract weight percentage in said composition is 0.006 %, and w/w ratio of the at least one peptide to apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[079] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00002 %, the apple fruit cellular extract weight percentage in said composition is 0.0006 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:30. [080] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00004 %, the apple fruit cellular extract weight percentage in said composition is 0.002 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:50.

[081] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; and (iii) contacting at least one peptide and apple fruit cellular extract to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00008 %, the apple fruit cellular extract weight percentage in said composition is 0.006 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:75.
[082] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) excipients; and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, and excipients to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80. [083] In an embodiment of the present disclosure, there is provided a method of preparing a composition to inhibit skin aging comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract, and suitable carriers, diluents and excipients to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:30.
[084] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising:

(i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients, and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, and excipients to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:50.
[085] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carrier; (d) diluent; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers (iv) obtaining diluent;(v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract, and suitable carriers, diluents, and excipients to obtain said composition, wherein w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:75.
[086] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; and (c) suitable carriers; (d) diluent; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; and (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract, suitable carriers, diluents, and excipients to obtain said composition, wherein said peptide is a tetrapetide having amino acid sequence Ile-Glu-Pro-Asp, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[087] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluent; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers (iv) obtaining diluents; (v) obtaining excipients, and (vi)

contacting at least one peptide, apple fruit cellular extract, suitable carriers, diluents, and excipients to obtain said composition, wherein the apple fruit cellular extract comprises chlorogenic acid having weight percentage in the said composition in the range of 0.0008 % - 0.0015 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[088] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; and (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluent; (v) obtaining excipients, and (vi) contacting at least one peptide, apple fruit cellular extract, suitable carrier, diluent, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%-0.0001%, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%- 0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[089] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carrier; (iv) obtaining diluent; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00002 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%- 0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.

[090] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (iv) contacting at least one peptide, apple fruit cellular extract, and suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00004 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%- 0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[091] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract, and suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00008 %, the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%- 0.007%, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[092] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers,

diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, the apple fruit cellular extract weight percentage in said composition is 0.0006 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[093] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, the apple fruit cellular extract weight percentage in said composition is 0.002 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.
[094] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluent; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (iv) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, excipients and combinations thereof to obtain said composition, wherein the at least one peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, the apple fruit cellular extract weight percentage in said composition is 0.006 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is in the range of 1:20 – 1:80.

[095] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00002 %, the apple fruit cellular extract weight percentage in said composition is 0.0006 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:30.
[096] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; and (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract and suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00004 %, the apple fruit cellular extract weight percentage in said composition is 0.002 %, and w/w ratio of the at least one peptide to the apple fruit cellular extract in said composition is 1:50. [097] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) at least one peptide; (b) apple fruit cellular extract; and (c) suitable carriers; (d) diluents; and (e) excipients, said method comprising: (i) obtaining at least one peptide; (ii) obtaining apple fruit cellular extract; (iii) obtaining suitable carriers; (iv) obtaining diluents; (v) obtaining excipients; and (vi) contacting at least one peptide, apple fruit cellular extract, suitable carriers, diluents, and excipients to obtain said composition, wherein the at least one peptide weight percentage in said composition is 0.00008 %, the apple fruit

cellular extract weight percentage in said composition is 0.006 %, and w/w ratio of the at least one peptide to apple fruit cellular extract in said composition is 1:75. [098] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20- 1:80; and (b) contacting the said composition with skin.
[099] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:30; and (b) contacting the said composition with skin.
[100] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:50; and (b) contacting the said composition with skin.
[101] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:75; and (b) contacting the said composition with skin.
[102] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20- 1:80, and said peptide is a tetrepeptide having

amino acid sequence Ile-Glu-Pro-Asp; and (b) contacting the said composition with skin.
[103] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20- 1:80, and the apple fruit cellular extract comprises chlorogenic acid having weight percentage in the range of 0.0008 % -0.0015 %; and (b) contacting said composition with skin.
[104] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20- 1:80, and the peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, and the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005% - 0.007%; and (b) contacting the said composition with skin.
[105] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80, the peptide weight percentage in said composition is 0.00002%, and apple fruit cellular extract weight percentage in said composition is in the range of 0.0005%- 0.007%; and (b) contacting the said composition with skin.
[106] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract,

wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20- 1:80, the peptide weight percentage in said composition is 0.00004%, and apple fruit cellular extract weight percentage in said composition is in the range of 0.0005% - 0.007%; and (b) contacting the said composition with skin.
[107] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80, the peptide weight percentage in said composition is 0.00008%, and the apple fruit cellular extract weight percentage in said composition is in the range of 0.0005% - 0.007%; and (b) contacting the said composition with skin.
[108] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80, the peptide weight percentage in said composition is in the range of 0.00001% - 0.0001%, and the apple fruit cellular extract weight percentage in said composition is 0.0006 %; and (b) contacting the said composition with skin.
[109] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80, the peptide weight percentage in said composition is in the range of 0.00001%- 0.0001%, and the apple fruit cellular extract

weight percentage in said composition is 0.002 %; and (b) contacting the said composition with skin.
[110] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide to apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80, the peptide weight percentage in said composition is in the range of 0.00001% - 0.0001%, and the apple fruit cellular extract weight percentage in said composition is 0.006 %; and (b) contacting the said composition with skin.
[111] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide weight percentage in said composition is 0.00002%, the apple fruit cellular extract weight percentage in said composition is 0.0006 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:30; and (b) contacting the said composition with skin.
[112] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide weight percentage in said composition is 0.00004%, the apple fruit cellular extract weight percentage in said composition is 0.002 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:50; and (b) contacting the said composition with skin.
[113] In an embodiment of the present disclosure, there is provided a method of inhibiting the appearance of skin aging, said method comprising: (a) obtaining a composition comprising: (i) at least one peptide; and (ii) apple fruit cellular extract, wherein the at least one peptide weight percentage in said composition is 0.00008%,

the apple fruit cellular extract weight percentage in said composition is 0.006 %, and the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is 1:30; and (b) contacting the said composition with skin.
[114] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one peptide; and (b) apple fruit cellular extract, wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 – 1:80, and said composition inhibits the appearance of skin aging by enhancing involucrin levels.
[115] 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
[116] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to be taken 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
[117] Preparation of Apple fruit callus from solid culture: Healthy Royal Gala Apple fruit were procured from local vendors and surface sterilized with 1% sodium hypochlorite for 10 minutes and 1% fungicide (Bavistin) for 10 minutes, followed by 0.05% mercuric chloride for 4 minutes. Apple fruits were washed with sterile water after each treatment under sterile conditions. Fruit tissues were collected and treated with antioxidant solution containing 200 mg PVP (polyvinyl phenol), 100 mg myo-inositol and 100 mg ascorbic acid in 1L sterile water for 5-10 minutes.
[118] The tissue was excised into pieces and inoculated on callus induction media: MS (Murashige and Skoog)- Salts with 1mg/L NAA, 3% sucrose as carbon source, 100 mg/L inositol, 200 mg/L PVP, 5 mg/L citric acid, gelling agar 0.7%, pH 5.8 and incubated at temperature 24±2 °C in the dark. Callus initiation was observed after 21 days.
[119] The callus induced from the apple fruit was excised and multiplied on callus multiplication media with the same ingredients except using MS- Salts with 2, 4-D in the place of NAA and without gelling agar.
[120] Preparation of apple fruit cell suspension culture: Apple fruit callus at 1-2 gm/100 ml of liquid media was weighed. This was then inoculated into liquid callus multiplication media comprising MS- Salts with 1mg/L 2, 4-D, 3% sucrose as carbon source, 100mg/L inositol, 200mg/L PVP, 5mg/L citric acid, pH 5.8. The cells were cultured at 120-140 RPM at a temperature of 24±2 °C without light and harvested after 28 days.
[121] Apple fruit cellular extract (ASC) preparation: Apple fruit stem cells were initially ground in a blender to make a fine paste. To this 0.3% (w/w) of sodium benzoate (preservative) and 0.2% (w/w) of L-ascorbic acid (anti-oxidant) were added and mixed thoroughly. The apple fruit stem cell paste was taken in to a freeze drying flask and was frozen using a 1:4 mixture of liquid nitrogen and acetone. The flask

was connected to a lyophilizer and the content was dried for 18 hours. The freeze dried fluffy off-whitish powder was obtained which was stored at -20 °C in an air-tight container.
Example 2
Biomarker analysis apple fruit cellular extract
[122] The apple fruit cellular extract so obtained was analysed for specific
biomarker standards. For this purpose, the amount of chlorogenic acid was analysed
using High Performance Liquid Chromatography (HPLC) (Figure 1). The
chlorogenic acid content in the lyophilised apple cellular extract was determined to
fall within the range of 0.0008% - 0.0015%
Example 3
Cellular cytotoxicity measurements of apple fruit cellular extract and peptide [123] Culture conditions: For analysing the cytotoxicity of apple fruit cellular extract and the peptide on HaCaT cells, HaCaT cells were grown in DMEM (Dulbecco's Modified Eagle Medium) media supplemented with 2 % FBS (Fetal Bovine Serum) and antibiotic-antimitotic solution in T75 flasks. The media was replaced every three days until more than 80 % confluency was obtained. The culture supernatant was tested for Mycoplasma contamination by MycoAlert Mycoplasma detection kit once every month.
[124] The cells were trypsinized with 0.25% trypsin EDTA solution for 4-5 minutes and sub-cultured in a ratio of 1:4 for scale-up for the assay. Wells of a 96 well plate were seeded with 3500 HaCaT cells in DMEM containing 2% FBS. The following day, the media was changed to M154 CF/PRF containing 30 µM CaCl2. After 5 days of incubation in a humidified incubator at 37°C at5% CO2, with one intermediate change of media on the third day, the designated wells were treated with the actives at the desired concentrations. The plates were incubated for a period of 24 hr in presence of the actives, following which the media was removed and the wells were

replaced with 200µL of media, containing 10% (20 µL) of Cell Titre Aqueous Solution (MTS Reagent). The plates were incubated at 37oC for a further 5-6 hr followed by reading at 490 nm using Spectramax Reader. To determine percent viability values, triplicate data points were analyzed and Average and SD values were calculated. Cytotoxicity of peptide (Table1), apple fruit cellular extract (Table 2) & L-citrulline (Table 3) is represented below. Table 1: Cytotoxicity result of apple fruit cellular extract

Table 2: Cytotoxicity results of peptide



[125] Cellular cytotoxicity to HaCaT cells at various weight percentages of apple fruit cellular extract was evaluated in order to determine optimum dosage levels for activity against inhibiting aging by enhancing levels of involucrin synthesis. As seen in Table 1, apple fruit cellular extract at various weight percentages (0.00002 -0.006%) does not significantly cause cellular toxicity. In particular, the cell extract at 0.00002 - 0.006% does not show any adverse effect on cell viability.
[126] Similar to Table 1, the effect of peptide at various weight percentages on cellular toxicity was also evaluated. As seen in Table 2, peptide in weight percentages in the range of 0.00004% - 0.0016% does not cause cellular toxicity. In particular, the peptide concentration range from 0.00004% - 0.00008 % does not show any adverse effect on cell viability.
[127] The effect of L-Citrulline, as a standard at various weight percentages on cellular toxicity was also analysed (Table 3). As seen from Table 3, L-Citrulline in weight percentage range of 0.008% - 0.25% does not cause cellular toxicity. In particular, peptide in the range of 0.008 % - 0.125 % does not show any adverse effect on cell viability.

[128] Based on the effect of apple fruit cellular extract and peptide on cellular toxicity as shown in Table 1 and Table 2, respectively, the concentrations of 0.00002 %, 0.00004 %, and 0.00008 % of peptide and 0.0006 %, 0.002 %, and 0.006 % of apple fruit cellular extract were chosen to test the effect of the composition on enhancing collagen synthesis.
Example 4
Effect of the combination of apple fruit cellular extract and peptide on
involucrin synthesis
[129] The effect of peptide at different weight percentages on the involucrin level
was analysed first in isolation (Figure 3). At the individual concentrations of 0.00004
%, 0.00008 %, and 0.00016 %, the tetrapeptide did not show any significant increase
in involucrin levels. In all these percentages highest production of involucrin
observed was of 59 % in case of the peptide with weight percentage of 0.00004 %.
Thus, it can be inferred that peptide alone does not affect the production of involucrin
significantly.
[130] As is in the previous case, the effect of various concentrations of apple fruit
cellular extract was also analyzed at different weight percentages (Figure 3). Apple
fruit cellular extract was analysed in isolation at 0.0006 %, 0.002 %, and 0.006 %.
The extract showed a greater ability than the tetrapeptide to increase involucrin
levels. The maximum production of involucrin was at 141%, and this was observed at
a weight percentage of 0.006 %.
[131] As apple fruit cellular extract showed some ability to increase involucrin
levels, the combination of the tetrapeptide and the apple fruit cellular extract was
attempted to determine whether this could enhance involucrin levels. Different w/w
ratios of peptide to apple fruit cellular extract analysed included 1:30 (0.00002 %
peptide + 0.0006 % apple fruit cellular extract), 1:50 (0.00004 % peptide + 0.002 %
apple fruit cellular extract) and 1:75 (0.00008 % peptide + 0.006 % apple fruit
cellular extract).

[132] Surprisingly, at all the ratios tried i.e. 1:30, 1:50 and 1:75, the combination of apple fruit cellular extract and peptide remarkably increased involucrin protein expression levels at 150 %, 147 %, and 158 % respectively (Figure 3). Maximum production of involucrin was observed at a ratio of 1:75. Thus, all the tried w/w ratios, i.e. 1:30, 1:50 and 1:75 of apple fruit cellular extract: peptide functions synergistically to enhance collagen synthesis at levels exceeding which the components by themselves could achieve.
[133] Further, it was observed that using the same concentrations of the peptide with the amino acid L-citrulline does not result in any synergism, rather, it results in the inhibition of the levels of involucrin protein obtained from L-citrulline. Thus, the composition of apple fruit cellular extract and the tetrapaeptide cannot be obtained without undue experimentation and is therefore not obvious to a person skilled in the art to arrive at.
[134] Overall, the present disclosure provides a composition comprising of apple fruit cellular extract and peptide that at different weight percentages and w/w ratios synergistically enhances involucrin synthesis. Further, at the particular w/w ratio of 1:30, 1:50 and 1:75, there is observed a synergistic increase in involucrin protein synthesis. As involucrin synthesis is associated with inhibiting skin aging symptoms such as appearance of wrinkles, and fine lines, enhanced involucrin synthesis, using a composition as described herein is likely to be useful in cosmetics which yield good results against appearance of fine lines and wrinkles associated with aging.

I/We Claim:
1. A composition to inhibit skin aging comprising:
(a) at least one peptide; and
(b) apple fruit cellular extract,
wherein the at least one peptide to the apple fruit cellular extract w/w ratio in said composition is in the range of 1:20 - 1:80.
2. The composition as claimed in claim 1, wherein the peptide is a tetrapeptide having amino acid sequence Ile-Glu-Pro-Asp.
3. The composition as claimed in claim 1, wherein the apple fruit cellular extract comprises chlorogenic acid having weight percentage in the range of 0.0008%-0.0015%.
4. The composition as claimed in claim 1, wherein the peptide w/w percentage in said composition is in the range of 0.00001% - 0.0001%, and apple fruit cellular extract w/w percentage in said composition is in the range of 0.0005% - 0.007%.
5. The composition as claimed in any of the claims 1-4, further comprising
suitable carriers, diluents, and excipients.
6. A method of preparing a composition as claimed in any of the claims 1-4, said
method comprising:
(a) obtaining at least one peptide;
(b) obtaining apple fruit cellular extract; and
(c) contacting the at least one peptide and the apple fruit cellular extract to obtain said composition.
7. A method of preparing a composition as claimed in claim 5, said method
comprising:
(a) obtaining at least one peptide;
(b) obtaining apple fruit cellular extract;
(c) obtaining suitable carriers, diluents, and excipients; and

(d) contacting the at least one peptide, the apple fruit cellular extract, suitable carriers, diluents, and excipients to obtain said composition.
8. A method of inhibiting the appearance of skin aging, said method comprising:
(a) obtaining a composition as claimed in any of the claims 1-5; and
(b) contacting said composition with skin.
9. The composition as claimed in claim 1, wherein said composition inhibits the
appearance of skin aging by enhancing involucrin levels.