FIELD OF INVENTION
The present disclosure broadly relates to the field of cosmetics and particularly refers to a composition proving anti-senescent effect.
BACKGROUND OF INVENTION
Cellular ageing or cellular senescence is a universal attribute of normal non-transformed cells that is recognized as one of the potential hallmarks of ageing. Cellular ageing is a process that is referred to as irreversible growth arrest of individual mitotic cells. The irreversible growth arrest of individual mitotic cells occurs in response to potentially oncogenic stimuli. These stimuli include damage to DNA, shortening and dysfunctioning of telomere, or oncogenic stress (Biomarkers of Cellular senescence and Skin aging; Audrey S. Wang).
The presence of senescent cells in skin has emerged as a potential contributor for skin ageing. It is suggested the number of senescent cells in the epidermal compartment of skin increases with age. The accumulation of senescent cells in skin with age are responsible for breakdown of extracellular matrix (ECM) (Quan T., Qin Z., Xia W., Shao Y., Voorhees J. J., Fisher G. J. (2009). Matrix-degrading metalloproteinases in photoaging. J. Investig. Dermatol. Symp. Proc. 14 20-24. 10.1038/jidsymp.2009.8). In addition, the presence of senescent cells affects skin elasticity and skin integrity contributing to ageing of skin.
Skin ageing leads to characteristic changes that are manifested in the form of fine lines and wrinkles on facial skin and other parts of the body.
Various strategies have been deployed to reduce the level of senescent cells in skin in order to combat skin ageing related problems. For instance: US20180002703A1 discloses a composition for reducing cellular senescence level. Also, discloses a method for reducing cellular senescence level inhibiting dcunld3 activity or inhibiting expression of dcunld3-encoding gene and use thereof.
AU2010233073B2 refers to methods of identifying, detecting a novel agent for preventing or treating an age-related disorder by improving mitochondrial function, maintaining the cell cycle- arrested state in senescent and post mitotic

cells. Further provides the use of low doses of rapamycin or its analogs in
preventing age-related diseases or disorders.
[007] Although a plethora of anti-skin agents have been described in literature to
enhance cellular activities, skin firmness and elasticity. However, the anti-agents
that are known in art do not provide much decrease in the senescence levels.
[008] Accordingly, there is a need for implementing a novel composition to
decrease cellular senescence in ageing cells.
SUMMARY OF THE INVENTION
[009] In an aspect of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4.
[0010] In another aspect of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition.
[0011] 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
[0012] 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.
[0013] Figure 1 depicts the graphical representation of cytotoxicity of ATP tested
on human dermal fibroblasts cells, in accordance with an embodiment of the present
disclosure.
[0014] Figure 2 depicts the graphical representation of cytotoxicity of Boerhavia
diffusa extract tested on human dermal fibroblasts cells, in accordance with an
embodiment of the present disclosure.
[0015] Figure 3 depicts the graphical representation of senescence associated beta
galactosidase activity of ATP and Boerhavia diffusa extract present at a weight ratio
of 1:1, in accordance with an embodiment of the present disclosure.
[0016] Figure 4 depicts the graphical representation of senescence associated beta
galactosidase activity of ATP and Boerhavia diffusa extract present at a weight ratio
of 1:0.2, in accordance with an embodiment of the present disclosure.
[0017] Figure 5 depicts the graphical representation of senescence associated beta
galactosidase activity of ATP and Boerhavia diffusa extract present at a weight ratio
of 1:5, in accordance with an embodiment of the present disclosure.
[0018] Figure 6 depicts the HPLC fingerprint of Boerhavia diffusa extract, in
accordance with an embodiment of the present disclosure.
[0019] Figure 7 depicts HPLC overlay of Boerhavia diffusa extract and gallic acid,
in accordance with an embodiment of the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0020] 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
[0021] 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.
[0022] 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.
[0023] 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”.
[0024] 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.
[0025] The term “including” is used to mean “including but not limited to”. “Including” and “including but not limited to” are used interchangeably. [0026] As used herein, the term “skin ageing” in the present disclosure refers to a multifactorial phenomenon exhibited by fine lines and wrinkles.
[0027] The term “senescence” as used herein is a process that refers to irreversible cell cycle arrest.
[0028] For the purpose of the present disclosure, senescence - associated beta-galactosidase refers to a hydrolase enzyme that catalyses the hydrolysis of beta-galactosides into monosaccharides in senescent cells.
[0029] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used

merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. [0030] 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.
[0031] 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.
[0032] A plethora of products that have been developed to address skin ageing problems failed to show substantial decrease in the cellular senescence. Further the ingredients that are utilized for preparing the said products associated with various side effects such as skin irritation, reduced skin elasticity, increased skin dryness. Also, the afore-mentioned products that are available in market are not cost effective.
[0033] Therefore, to overcome the afore-mentioned problems, the object of the present disclosure is to provide a composition addressing skin ageing by reducing the senescence levels, wherein the composition is based on natural extracts. The object of the present disclosure further discloses a composition comprising at least one purine nucleic acid substance; and Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4. The combination of at least one purine nucleic acid substance; and Boerhavia diffusa extract provides a synergistic effect in decreasing enzymatic levels of senescence associated beta galactosidase. The decrease in senescence associated beta galactosidase enzyme levels leads to the enhancement of cellular activities, skin firmness and elasticity.

[0034] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4. In another embodiment of the present disclosure, the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.45 to 1:4. In yet another embodiment of the present disclosure, the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.475 to 1:3.5
[0035] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof. [0036] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).
[0037] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).

[0038] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4. In another embodiment of the present disclosure, the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3. In yet another embodiment of the present disclosure, the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.65 to 1:2.5.
[0039] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof. [0040] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).
[0041] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).

[0042] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4. In another embodiment of the present disclosure, the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2. In yet another embodiment of the present disclosure, the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio of 1:1. [0043] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof. [0044] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2 and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).
[0045] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).

[0046] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4, and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition. In another embodiment of the present disclosure, the at least one purine nucleic acid substance has a weight percentage in a range of 0.002-0.09% with respect to the composition. In yet another embodiment of the present disclosure, the at least one purine nucleic acid substance has a weight percentage in a range of 0.006-0.03% with respect to the composition. [0047] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3, and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition
[0048] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition.
[0049] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine

5’-triphosphate (ATP) and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0050] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0051] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition.
[0052] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0053] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the

Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0054] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition. [0055] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition. In another embodiment of the present disclosure, the Boerhavia diffusa extract has a weight percentage in a range of 0.009-0.1% with respect to the composition. In yet another embodiment of the present disclosure, the Boerhavia diffusa extract has a weight percentage in a range of 0.009-0.03% with respect to the composition.
[0056] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and

wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition.
[0057] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition. [0058] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition. [0059] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-

triphosphate (ATP) and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition.
[0060] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition.
[0061] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.02% with respect to the extract.
[0062] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition, and wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.02% with respect to the extract.
[0063] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1%

with respect to the composition, and wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.02% with respect to the extract.
[0064] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2 and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition, and wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.02%% with respect to the extract.
[0065] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition.
[0066] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).

[0067] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).
[0068] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).
[0069] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to

obtain the composition and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition.
[0070] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4, said process comprising: (i) obtaining the at least one purine nucleic acid substance; (ii) obtaining the Boerhavia diffusa extract; and (iii) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition and wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.21% with respect to the extract.
[0071] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the composition decreases production of senescence associated beta galactosidase. [0072] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof and wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP) and wherein the composition decreases production of senescence associated beta galactosidase.
[0073] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia

diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3 and wherein the composition decreases production of senescence associated beta galactosidase. [0074] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition and wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition and wherein the composition decreases production of senescence associated beta galactosidase.
[0075] In an embodiment of the present disclosure, there is provided a composition comprising: (a) at least one purine nucleic acid substance; and (b) Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4 and wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.02% with respect to the extract and wherein the composition decreases production of senescence associated beta galactosidase.
[0076] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[0077] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and

compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply. The working and non-working examples as depicted in the forthcoming sections highlight the criticality of the working percentages of different components in achieving the composition of the present disclosure. It is further specified that the presence of at least one purine nucleic acid substance and Boerhavia diffusa extract, present at a weight ratio range of 1:0.4 to 1:4 is critical so as to provide synergistic decrease in production of senescence associated beta glucosidase. The absence of any of the components specified above or variation in the weight ratio range substantially affects the synergistic effect in decreasing production of senescence associated beta glucosidase.
Materials and Methods
[0078] The components of the present disclosure: ATP having catalogue no. A2383 was procured from sigma, and Boerhavia diffusa extract was commercially procured. from Amrutha herbals. The stock of ATP was prepared in water, wherein pH was adjusted with 1N NaOH and stock of Boerhavia diffusa extract was prepared in water. Human dermal fibroblast (HDF) cells were procured from LONZA, fibroblast basal medium was procured from INVITROGEN, cellular senescence assay kit was procured from CellBiolabs, and 10X phosphate buffered saline was procured from Sigma.
[0079] The characterization and identification of active present in Boerhavia diffusa extract, was achieved by High-Performance Liquid Chromatography (HPLC) analyses. The following conditions were required for carrying out HPLC analysis (as depicted in Figure 6) of Boerhavia diffusa extract: Column: Inertsil ODS 3V 250mm X4.6mm, 5µ; flow rate: 1.0mLl/min; Wavelength: 270nm; Mobile phase: 0.1% H3PO4 in water: Acetonitrile (70:30); Column temperature: 250℃. From the analysis of HLPC profile of Boerhavia diffusa extract, it was found that Boerhavia diffusa extract comprised gallic acid having a weight percentage of 0.01% with respect to the extract.
Example 1

Evaluation of cytotoxicity for ATP and Boerhavia diffusa extract
[0080] ATP and Boerhavia diffusa extract was utilized at a weight percentage
ranging from 0.001-0.1% and 0.005-0.1% respectively, in order to determine the
cytotoxicity level. The cytotoxicity level was determined in vitro by using WST-1
Cell Proliferation and Cell Viability kit from Clontech.
[0081] Figure 1 depicts the cytotoxicity of ATP. It can be observed from Figure 1
that cells treated with 0.025% of ATP exhibited 92% cell survival as compared to
control.
[0082] Figure 2 depicts the cytotoxicity of Boerhavia diffusa extract, wherein it
can be observed that cells treated with 0.025% of Boerhavia diffusa extract
exhibited 105% cell survival as compared to control.
[0083] Therefore, it can be inferred from Figure 1 and Figure 2 that ATP and
Boerhavia diffusa extract was safe to use, wherein the cells that were treated with
0.025% of ATP and 0.025% of Boerhavia diffusa extract showed maximum
survival rate of 92% and 105%, respectively. The ATP and Boerhavia diffusa
extract having a weight percentage range of 0.001-0.1% and 0.005-0.1% was used
further for the purpose of the present disclosure.
Example 2
Method for evaluating effect of the composition on expression of senescence
associated beta galactosidase enzyme
[0084] The effect of the composition on expression of senescence associated beta
galactosidase enzyme was evaluated by cellular senescence assay. The cellular
senescence assay was carried out for a period of 3 days and consisted of following
steps:
[0085] Step 1: HDF cells were seeded in 96- well plates and incubated in 5% CO2
incubator on day 1.
[0086] Step 2: Adherent HDF cells were exposed to 100µM Hydrogen peroxide
(H2O2) for 3hours, that was followed by treatment with actives (ATP and
Boerhavia diffusa extract) in serum free fibroblast basal medium for 24 hrs. on day
2.

[0087] Step 3: Post incubation with actives, cells were washed twice with 1X PBS on day 3. Cell Lysis and processing was performed as per the manufacturer’s protocol (CELLBIOLABS) to estimate the enzymatic levels of senescence associated beta galactosidase (SA-β-gal) expression.
Example 3
Evaluation of effect of ATP and Boerhavia diffusa extract present at a weight
ratio of 1:1
[0088] The combination of ATP and Boerhavia diffusa extract present at a
concentration of 0.01% each, were used to evaluate the enzymatic levels of
senescence associated beta galactosidase (SA-β-gal) expression.
[0089] Figure 3 depicts the effect of combination of 0.01% of ATP and 0.01% of
Boerhavia diffusa extract on SA-β-gal expression levels. It was observed from
Figure 3 that the combination of 0.01% of ATP and 0.01% of Boerhavia diffusa
extract synergistically decreased SA-β-gal expression levels responsible for cellular
senescence. It can be deduced from Figure 3 that the synergistic decrease in SA-β-
gal expression levels helped in enhancing cellular activities in ageing skin.
[0090] It can be inferred that combination of 0.01% of ATP and 0.01% of
Boerhavia diffusa extract was present within the disclosed weight range of the
present disclosure, i.e. 1:0.4-1:4. Further, said combination provided synergistic
effect in decreasing SA-β-gal expression levels. Therefore, combination of 0.01%
of ATP and 0.01% of Boerhavia diffusa extract were considered as the working
example of the present disclosure.
Example 4
Evaluation of effect of ATP and Boerhavia diffusa extract present at a weight
ratio of 1:0.2
[0091] Combination having 0.1% of ATP and 0.02% of Boerhavia diffusa extract
was used to evaluate the enzymatic levels of SA-β-gal expression. Referring to
Figure 4, it can be observed that 0.1% of ATP and 0.02% of Boerhavia diffusa
extract did not show any substantial decrease in SA-β-gal expression.
[0092] It is clear from Figure 4 that the combination of 0.1% of ATP and 0.02%
of Boerhavia diffusa extract was present outside the disclosed weight ratio range of

the present disclosure. Further, the combined effect of 0.1% of ATP and 0.02% of Boerhavia diffusa extract was less than that expected by additive effect 0.1% of ATP and 0.02% of Boerhavia diffusa extract.
[0093] It can be inferred that combination of 0.1% of ATP and 0.02% of Boerhavia diffusa extract did not exhibit enhanced decrease in SA-β-gal expression. Therefore, said combination at weight ratio of 1:0.2 were considered as non-working example of the present disclosure.
Example 5
Evaluation of effect of ATP and Boerhavia diffusa extract present at a weight
ratio of 1:5
[0094] The combination of ATP and Boerhavia diffusa extract was utilized at a
concentration of 0.001% and 0.005% respectively for the purpose of the present
disclosure.
[0095] Referring to Figure 5, it can be observed that the combination having
0.001% ATP and 0.005% of Boerhavia diffusa extract did not show any substantial
decrease in SA-β-gal expression. It can also be observed that the afore-mentioned
combination was present outside the working range of the present disclosure.
[0096] It is clear from Figure 5 that the combined effect of 0.001% of ATP and
0.005% of Boerhavia diffusa extract was less than that expected by additive effect
of 0.001% of ATP and 0.005% of Boerhavia diffusa extract. Therefore,
combination of 0.001% of ATP and 0.005% of Boerhavia diffusa extract that did
not exhibit enhanced decrease in SA-β-gal expression and was considered as the
non-working example of the present disclosure.
[0097] Based on the observations made from the analysis of HPLC profile (as
depicted in Figure 6) of Boerhavia diffusa extract carried out under the conditions
(as mentioned in materials and methods of the present disclosure), it was observed
that gallic acid was present as active in Boerhavia diffusa extract, wherein
Boerhavia diffusa extract was having gallic acid content of about 0.2ppm, as
depicted in Figure 7.
[0098] Overall, the combination of ATP and Boerhavia diffusa extract in the
disclosed weight ranges is essential to have a synergistic effect in decreasing

senescence-associated beta galactosidase (SA-β-gal) enzyme synthesis. Deviating ATP concentration or Boerhavia diffusa extract concentration from the disclosed weight ranges does not exhibit the desired result.
Advantages of the present disclosure
[0099] The present disclosure discloses a composition comprising at least one purine nucleic acid substance; and Boerhavia diffusa extract, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4. The composition provides synergistic effect in decreasing the enzymatic levels of senescence-associated beta galactosidase (SA-β-gal) enzyme expression. The composition involving natural extract helps in enhancing the cellular activities, skin firmness and elasticity in ageing skin without producing any side effects. Therefore, the composition is safe to use. Also, the composition is cost-effective.

I/We Claim:
1. A composition comprising:
a) at least one purine nucleic acid substance; and
b) Boerhavia diffusa extract,
wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.4 to 1:4.
2. The composition as claimed in claim 1, wherein the at least one purine nucleic acid substance is selected from a group consisting of adenosine 5’-triphosphate (ATP), adenosine 2′-monophosphate, adenosine 3′-monophosphate, adenosine 5′-monophosphate, and salts thereof.
3. The composition as claimed in claim 2, wherein the at least one purine nucleic acid substance is disodium salt hydrate of adenosine 5’-triphosphate (ATP).
4. The composition as claimed in claim 1, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.5 to 1:3.
5. The composition as claimed in claim 1, wherein the at least one purine nucleic acid substance to the Boerhavia diffusa extract has a weight ratio in a range of 1:0.7 to 1:2.
6. The composition as claimed in claim 1, wherein the at least one purine nucleic acid substance has a weight percentage in a range of 0.001-0.1% with respect to the composition.
7. The composition as claimed in claim 1, wherein the Boerhavia diffusa extract has a weight percentage in a range of 0.005-0.1% with respect to the composition.
8. The composition as claimed in claim 1, wherein the Boerhavia diffusa extract comprises gallic acid having a weight percentage in a range of 0.005-0.02% with respect to the extract.
9. A process for preparing the composition as claimed in claim 1, said process comprising:
a) obtaining the at least one purine nucleic acid substance;

b) obtaining the Boerhavia diffusa extract; and
c) contacting the at least one purine nucleic acid substance and the Boerhavia diffusa extract, to obtain the composition.
10. The composition as claimed in any one of the claims 1-8, wherein the composition decreases production of senescence associated beta galactosidase.