FIELD OF INVENTION
[001] The present disclosure broadly relates to field of cosmetics, and particularly relates to a composition comprising aloin and caffeic acid for addressing skin aging.
BACKGROUND OF INVENTION
[002] The matrix metalloprotease are zinc-containing endopeptidases which mediate the degradation of the different components of the extracellular matrix secretions of the cells. Matrix metalloprotease 1 (MMP1) also known as interstitial collagenase is the protease involved in the breakdown of collagen, the predominant protein of the ECM. These proteases are secreted by keratinocytes and dermal fibroblasts in response to multiple stimuli such as aging, oxidative stress, UV radiation, and cytokines. Upon the onset of skin aging the formation of fine lines and wrinkles appear on the facial skin and other parts of the body due to fragmented collagen as a result of the up regulated MMP1 levels. The decrease in the density of the collagen protein in the ECM results in the loss of the skin's mechanical properties as a result of which skin loses its firmness and elasticity. [003] As a result of aging, numerous dynamic changes tend to happen to the skin which is a manifestation of molecular pathways. Therefore, for a composition to be effective it needs to act on a proper target through which it is able to exert its effect on the skin. Many compositions available make use of synthetic components which has high risks of developing side effects.
[004] CA2857231A1 discloses a skin anti-aging composition which comprises myricetin, or glycoside thereof, and at least one LXR alpha agonist. [005] EP0688559 Al discloses anti-aging compositions for preventing cutaneous aging. The composition comprises one or two of aminoethyl compounds represented by the formula as described.
[006] In light of lack of available compositions that target the basic molecular pathways, novel compositions are constantly sought after to fill the void created in the field.

SUMMARY OF THE INVENTION
[007] In an aspect of the present disclosure, there is provided a composition comprising: (a) aloin; and (b) caffeic acid, wherein aloin to caffeic acid has a w/w ratio in a range of 8:1 to 12:1.
[008] In an aspect of the present disclosure, there is provided a process for preparing the composition comprising: (a) aloin; and (b) caffeic acid, wherein aloin to caffeic acid w/w ratio is in a range of 8:1 to 12:1, said process comprising: (i) obtaining aloin; (ii) obtaining caffeic acid; and (iii) contacting aloin and caffeic acid, to obtain the composition.
[009] 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
[0010] 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.
[0011] Figure 1 illustrates the cytotoxicity assay for aloin, in accordance with an
embodiment of the present disclosure.
[0012] Figure 2 illustrates the cytotoxicity assay for caffeic acid, in accordance with
an embodiment of the present disclosure.
[0013] Figure 3 illustrates the effect of aloin (0.01%), and caffeic acid (0.001%), in
accordance with an embodiment of the present disclosure.

[0014] Figure 4 illustrates the effect of aloin (0.01%), and caffeic acid (0.01%), in accordance with an embodiment of the present disclosure.
[0015] Figure 5 illustrates the effect of aloin (0.01%), and caffeic acid (0.0005%), in accordance with an embodiment of the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0016] Those skilled in the art will be aware that the present disclosure is subject to
variations and modifications other than those specifically described. It is to be
understood that the present disclosure includes all such variations and
modifications. The disclosure also includes all such steps, features, compositions,
and compounds referred to or indicated in this specification, individually or
collectively, and any and all combinations of any or more of such steps or features.
Definitions
[0017] For convenience, before further description of the present disclosure,
certain terms employed in the specification, and examples are delineated here.
These definitions should be read in the light of the remainder of the disclosure and
understood as by a person of skill in the art. The terms used herein have the
meanings recognized and known to those of skill in the art, however, for
convenience and completeness, particular terms and their meanings are set forth
below.
[0018] The articles "a", "an" and "the" are used to refer to one or to more than one
(i.e., to at least one) of the grammatical object of the article.
[0019] The terms "comprise" and "comprising" are used in the inclusive, open
sense, meaning that additional elements may be included. It is not intended to be
construed as "consists of only".
[0020] Throughout this specification, unless the context requires otherwise the
word "comprise", and variations such as "comprises" and "comprising", will be
understood to imply the inclusion of a stated element or step or group of element or
steps but not the exclusion of any other element or step or group of element or
steps.

[0021] The term "including" is used to mean "including but not limited to". "Including" and "including but not limited to" are used interchangeably. [0022] Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. [0023] Aloin is an anthraquinone glycoside isolated from Aloe species plants. It is a lemon yellow coloured crystalline compound known for various health benefits. [0024] Caffeic acid (3,4-dihydroxy-cinnamic acid) is an organic compound, is a potent antioxidant, and can be naturally found in many plants. It has also been shown to have anti-inflammatory effects.
[0025] For the purposes of the present document, 'skin aging' refers to the manifestation of aging process that is visible on the skin in form of wrinkles or facial lines.
[0026] As discussed previously, the decrease in the density of the collagen protein in the ECM results in the loss of the skin's mechanical properties as a result of which skin loses its firmness and elasticity. Literature survey (Pittayapruek P., 2016. Int. Mol Sci. 17(6); MinaYaar, et al., 2002. J.Invest. Dermatol.Symp.Proc; 7:51 -58; and Xia et al., 2013 Aging Cell.; 12(4):661-71) reveals that active research is carried out to decrease the levels of MMP1 by inhibiting the MMP1 synthesis which can eventually decrease facial fine lines and wrinkles. The present composition being herbal in nature is also devoid of any side effects. [0027] The present disclosure aims to address the issue of providing an anti-aging composition by targeting the production of MMP-1 enzyme production. The present disclosure provides a composition comprising aloin and caffeic acid in a w/w ratio having a range of 8:1 to 12:1 (aloin to caffeic acid). The composition has been shown to synergistically inhibit the production of MMP-1 enzyme production. [0028] An excipient is an inactive substance that serves as the vehicle or medium for a drug or other active substance. Excipients include colouring agents,

humectants, diluting agent, preservatives, emollients, other known cosmetically
suitable additives, and combinations thereof.
[0029] 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.
[0030] The present disclosure is not to be limited in scope by the specific
embodiments described herein, which are intended for the purposes of
exemplification only. Functionally-equivalent products, compositions, and methods
are clearly within the scope of the disclosure, as described herein.
[0031] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) aloin; and (b) caffeic acid, wherein aloin to caffeic acid has a w/w
ratio in a range of 8:1 to 12:1. In another embodiment of the present disclosure,
aloin to caffeic acid has a w/w ratio in a range of 9:1 to 11:1.
[0032] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) aloin; and (b) caffeic acid, wherein aloin to caffeic acid has a w/w
ratio of 10:1.
[0033] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) aloin having a weight percentage in a range of 0.005-0.015% with
respect to the composition; and (b) caffeic acid having a weight percentage in a
range of 0.0005-0.002% with respect to the composition, wherein aloin to caffeic
acid has a w/w ratio in a range of 8:1 to 12:1. In another embodiment of the present
disclosure, aloin has a weight percentage in a range of 0.007-0.012%) with respect
to the composition, and caffeic acid has a weight percentage in a range of 0.0007-
0.0015%) with respect to the composition.
[0034] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) aloin having a weight percentage of 0.01% with respect to the
composition; and (b) caffeic acid having a weight percentage of 0.001% with
respect to the composition, wherein aloin to caffeic acid has a w/w ratio of 10:1.

[0035] In an embodiment of the present disclosure, there is provided a composition comprising: (a) aloin; (b) caffeic acid; and (c) at least one excipient, wherein the at least one excipient is selected from the group consisting of water, DMSO, polymer, gum base, chelating agent, emulsifier, surfactant, bulking agent, humectant, solvent, antimicrobial agent, moisturizer, fragrance, and combinations thereof. Wherein aloin to caffeic acid has a w/w ratio in a range of 8:1 to 12:1. [0036] In an embodiment of the present disclosure, there is provided a composition comprising: (a) aloin; (b) caffeic acid; and (c) at least one excipient, wherein the at least one excipient is selected from the group consisting of water, dimethyl sulfoxide (DMSO), polymer, gum base, chelating agent, emulsifier, surfactant, bulking agent, humectant, solvent, antimicrobial agent, moisturizer, fragrance, and combinations thereof, wherein aloin to caffeic acid has a w/w ratio of 10:1. [0037] In an embodiment of the present disclosure, there is provided a composition comprising: (a) aloin having a weight percentage in the range of 0.005% - 0.015% with respect to the composition; (b) caffeic acid having a weight percentage in the range of 0.0005%) - 0.002%) with respect to the composition; (c) polymers having a weight percentage in the range of 0.2 - 0.5% with respect to the composition; (d) gum base having a weight percentage in the range of 0.1 - 0.5% with respect to the composition; (e) chelating agents having a weight percentage in the range of 0.05 -0.2%) with respect to the composition; (f) emulsifiers having a weight percentage in the range of 1 - 5% with respect to the composition; (g) surfactants having a weight percentage in the range of 0.5 - 3% with respect to the composition; (h) humectants having a weight percentage in the range of 0.1 - 1% with respect to the composition; (i) solvents having a weight percentage in the range of 1 - 3% with respect to the composition; (j) antimicrobial agents having a weight percentage in the range of 0.1 - 1% with respect to the composition; (k) moisturizers having a weight percentage in the range of 2 - 3% with respect to the composition; and (1) bulking agent having a weight percentage in a range of 0.5-5% with respect to the composition, wherein aloin to caffeic acid has a w/w ratio in a range of 8:1 to 12:1. [0038] In an embodiment of the present disclosure, there is provided a composition comprising: (a) aloin having a weight percentage in the range of 0.005%) - 0.015%)

with respect to the composition; (b) caffeic acid having a weight percentage in the range of 0.0005% - 0.002% with respect to the composition; (c) polymers having a weight percentage in the range of 0.3 - 0.4% with respect to the composition, and is selected from the group consisting of ammonium acryloyl - dimethyltaurate, Cio-30 alkyl acrylate crosspolymer, sodium acryloyldimethyl taurate copolymer (and) isohexadecane (and) polysorbate 60, pectin, and combinations thereof; (d) gum base having a weight percentage in the range of 0.2 - 0.4% with respect to the composition, and is selected from the group consisting of xanthan gum, guar gum, and combinations thereof; (e) chelating agents having a weight percentage in the range of 0.1 - 0.15% with respect to the composition, and is selected from the group consisting of disodium EDTA, tetrasodium EDTA, sodium phosphate, calcium disodium EDTA, and combinations thereof; (f) emulsifiers having a weight percentage in the range of 2 - 4% with respect to the composition, and is selected from the group consisting of potassium cetyl phosphate (and) hydrogenated palm glycerides, cetostearyl alcohol, magnesium isodecylbenzenesulfonate, glyceryl stearate (and) PEG-100 stearate, polysorbates, laureth-4, and potassium cetyl sulfate, and combinations thereof; (g) surfactants having a weight percentage in the range of 1 - 2.5% with respect to the composition, and is selected from the group consisting of triethanolamine, coconut oil, sodium laureth sulfate, and combinations thereof; (h) humectants having a weight percentage in the range of 0.3 - 0.7% with respect to the composition, and is selected from the group consisting of ethylhexyl glycerin, propylene glycol, glycerine, sorbitol, and combinations thereof; (i) solvents having a weight percentage in the range of 1.5 - 2.5% with respect to the composition, and is selected from the group consisting of propylene carbonate, stearate oil, mineral oil, and combinations thereof; (j) antimicrobial agents having a weight percentage in the range of 0.3 - 0.7% with respect to the composition, and is selected from the group consisting of phenoxyethanol, parabens, and combinations thereof; (k) moisturizers having a weight percentage in the range of 2.3 - 2.7% with respect to the composition, and is selected from the group consisting of C12-15 alkyl benzoate, hyaluronic acid, essential oils, and combinations thereof; (1) bulking agent having a

weight percentage in the range of 0.5-5% with respect to the composition, and is selected from the group consisting of magnesium sulfate, magnesium myristate, zinc oxide, silica, and combinations thereof, wherein aloin to caffeic acid w/w ratio is in a range of 12:1 to 8:1.
[0039] In an embodiment of the present disclosure, there is provided a composition comprising: (a) aloin having a weight percentage in a range of 0.005-0.015% with respect to the composition; (b) caffeic acid having a weight percentage in a range of 0.0005-0.002%) with respect to the composition; (c) ammonium acryloyl -dimethyltaurate having a weight percentage of 0.05% with respect to the composition; (d) C10-30 alkyl acrylate crosspolymer having a weight percentage of 0.17%) with respect to the composition; (e) xanthan gum having a weight percentage of 0.10%> with respect to the composition; (f) disodium EDTA having a weight percentage of 0.10%> with respect to the composition; (g) magnesium sulfate having a weight percentage of 0.70% with respect to the composition; (h) C12-15 alkyl benzoate having a weight percentage of 3% with respect to the composition; (i) potassium cetyl phosphate (and) hydrogenated palm glycerides having a weight percentage of 2% with respect to the composition; (j) cetostearyl alcohol having a weight percentage of 1.50% with respect to the composition; (k) glyceryl stearate (and) PEG-100 stearate having a weight percentage of 2% with respect to the composition; (1) triethanolamine having a weight percentage of 0.2% with respect to the composition; (m) sodium acryloyl dimethyl taurate copolymer (and) isohexadecane (and) polysorbate 60 having a weight percentage of 3% with respect to the composition; (n) phenoxyethanol having a weight percentage of 0.6% with respect to the composition; (o) ethylhexyl glycerin having a weight percentage of 0.5%) with respect to the composition; and (p) fragrance having a weight percentage of 0.8%) with respect to the composition, and wherein aloin to caffeic acid w/w ratio is in a range of 8:1 to 12:1.
[0040] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) aloin; and (b) caffeic acid, wherein aloin to caffeic acid weight ratio is in a range of 8:1 to 12:1, said process comprising: (i)

obtaining aloin; (ii) obtaining caffeic acid; and (iii) contacting aloin and caffeic
acid, to obtain the composition.
[0041] In an embodiment of the present disclosure, there is provided a process for
preparing the composition comprising: (a) aloin; and (b) caffeic acid, wherein aloin
to caffeic acid weight ratio is 10:1, said process comprising: (i) obtaining aloin; (ii)
obtaining caffeic acid; and (iii) contacting aloin and caffeic acid, to obtain the
composition.
[0042] In an embodiment of the present disclosure, there is provided a process for
preparing the composition comprising: (a) aloin having a weight percentage in a
range of 0.005-0.015% with respect to the composition; and (b) caffeic acid having
a weight percentage in a range of 0.0005-0.002% with respect to the composition,
wherein aloin to caffeic acid weight ratio is in a range of 8:1 to 12:1, said process
comprising: (i) obtaining aloin; (ii) obtaining caffeic acid; and (iii) contacting aloin
and caffeic acid, to obtain the composition.
[0043] In an embodiment of the present disclosure, there is provided a process for
preparing the composition comprising: (a) aloin; (b) caffeic acid; and (c) at least
one excipient, wherein aloin to caffeic acid weight ratio is in a range of 8:1 to 12:1,
said process comprising: (i) obtaining aloin; (ii) obtaining caffeic acid; (iii)
obtaining the at least one excipient; and (iv) contacting aloin, caffeic acid, and the
at least one excipient, to obtain the composition, wherein the at least one excipient
is selected from the group consisting of water, dimethyl sulfoxide (DMSO),
polymer, gum base, chelating agent, emulsifier, surfactant, bulking agent,
humectant, solvent, antimicrobial agent, moisturizer, fragrance, and combinations
thereof.
[0044] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein the composition inhibits MMP1 (matrix
metalloprotease 1) protein synthesis.
[0045] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein the composition reduces skin wrinkles caused due to
aging.

[0046] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein said composition can be for topical application. [0047] 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. [0048] 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. [0049] 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. [0050] 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. [0051] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[0052] 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. [0053] The working and non-working examples of the composition comprising aloin, and caffeic acid will provide support to the weight percentage range and weight ratio as claimed in the present disclosure. The effect of the present composition on inhibition of MMP-1 protein production will be depicted in the forthcoming sections. Material and Methods

[0054] Aloin and caffeic acid were procured from Sigma. Stock solution was prepared in DMSO.
Protocol of experiments performed
[0055] Human dermal fibroblasts were seeded in 12- well plates (Costar) in Fibroblast basal media (LONZA). The culture was incubated in 5% CO2 incubator and was allowed to reach 100% confluency. Then the fully confluent fibroblast cells are treated with actives and incubated in 5% CO2 incubator. Spent medium was harvested after 48 hrs.
[0056] MMP1 ELISA was performed as per manufacturer's protocol to estimate secreted MMP1. Briefly, plates were thawed on ice on the day MMP-1 estimation was performed using Human MMP-1 ELISA Kit, (Sigma, and Cat # RAB0361). Before starting the ELISA assay, all the kit components were allowed to reach room temperature. Samples were diluted by 100-fold in 96-wells deep well block by transferring 5ul of sample in 495 ul of IX assay diluent buffer supplied with the kit. The contents were mixed thoroughly to ensure uniformity of the suspension. Lyophilized Human MMP-1 Protein Standard vial supplied with the kit was briefly centrifuged and 400 uL of IX Assay Diluent was added and mixed to prepare a 0.1 25 ug/mL standard. 5 ul of 0.1 ug/mL MMP-1 standard was added to 995 uL of IX assay diluent to achieve 500 pg/mL concentration. 2-fold serial dilution from 500 pg/ml stock was carried out in assay diluent buffer upto 7.8 pg/ml for the standard curve. lOOul of samples (n=3) and MMP-1 standard dilutions (n=4) along with lOOul of only. Assay diluent (which serves as blank) was transferred to Human MMP-1 Antibody-coated ELISA plate and incubated for 2.5 hours at room temperature with gentle shaking. Meanwhile, the Biotinylated Detection Antibody vial was briefly centrifuged and 100 uL of lx Assay Diluent Buffer was added to prepare a detection antibody concentrate (80x). The detection antibody concentrate was then diluted by 80-fold with lx assay diluent buffer for final use. 20x Wash Buffer was diluted to obtain adequate volume of lx wash buffer. Samples/solution from the ELISA plates were decanted out and washed 4 times with lx wash buffer by filling each well with 300 ul of wash buffer using a multichannel pipette.

Complete removal of liquid at each step was made sure by blot-drying the plates. 100 ul of lx prepared Biotinylated Detection Antibody was added to each well and the plates were incubated for 1 hour at room temperature with gentle shaking. After incubation, the detection antibody solution was discarded from the plate and the wash step was repeated with lx wash buffer. The plates were dried by tapping against paper-towel. The HRP-Streptavidin concentrate vial was briefly spun and pipetted up and down to mix gently before use, as precipitates may form during storage. HRP-Streptavidin concentrate was then diluted 440-fold with lx Assay Diluent Buffer before use. lOOul of prepared lx HRP-Streptavidin solution was then added to each well and incubated for 45 minutes at room temperature with gentle shaking. After incubation, the HRP-Streptavidin solution was discarded and the wash step was repeated with lx wash buffer and the plates were blot-dried. lOOul of ELISA colorimetric TMB Reagent was added to each well and plates were incubated for 30 minutes at room temperature (25±2°C) in dark with gentle shaking. 50ul of stop solution was added to each well and the absorbance was measured at 450 nm immediately.
[0057] Cell viability assay: Fibroblast (American type cell Culture, India) were cultured in DMEM medium (Sigma, India). For experiments, confluent cells were trypsinized and suspended in DMEM growth medium at 2 x 105 cells/ml. Then the cells were placed in 96-well plates (2 x 104 cells/well) for cell viability assay. Stock solution of aloin and caffeic acid was prepared in DMSO. After 24 hours, the cells were treated with aloin and caffeic acid respectively at 37°C and incubated for 72 hours. The media was then removed and 20uL of 5mg/ml solution of 15 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was added to each well. The formazan crystals were solubilized following addition of lOOul of dimethyl sulfoxide and absorbance was read at 540nm.
Example 1
Cytotoxicity of aloin and caffeic acid
[0058] The present example evaluated the cytotoxicity of the actives i.e. aloin and
caffeic acid.

[0059] The assay was performed as per the protocol mentioned in previous section.
Figure 1 depicts the cytotoxicity assay for aloin over a concentration of 0.001-
0.5%. Referring to Figure 1, it can be observed that the viability of the fibroblast
cells was not affected till a concentration of 0.01%. Around 85% of the cells were
viable till a concentration of 0.5%.
[0060] Figure 2 depicts the cytotoxicity assay for caffeic acid over a concentration
of 0.001%-0.05%. It can be inferred from Figure 2, that the viability of the
fibroblast cells was not affected till a concentration of 0.01%. Significant reduction
in viability was observed for higher concentration of caffeic acid.
[0061] Therefore, the cytotoxicity assay depicts the safe concentration of the
actives which can be used for preparing the composition for skin benefits.
Example 2
Effect of aloin (0.01%) and caffeic acid (0.001%) on MMP1 protein expression [0062] The present example studies the effect of aloin (0.01%) and caffeic acid (0.001%) on MMP1 protein expression.
[0063] Figure 3 depicts the inhibition of MMP1 protein inhibition in the presence of the composition. It can be appreciated that caffeic acid and aloin in isolation are not able to inhibit the production of MMP1 production, but at a combination of aloin (O.OP/o) and caffeic acid (0.001%) a synergistic inhibition can be observed. A reduction of around 18.5% (81.5% MMP1 protein production) can be observed (Figure 3) in the presence of the present composition. Therefore, the composition comprising aloin and caffeic acid in a weight ratio of 10:1 is showing a synergistic effect in inhibiting the MMP1 protein production. The composition of the present disclosure helps in preventing collagen breakdown thereby increasing the tensile strength of the skin, helping in reduction of facial fine lines and wrinkles
Example 3
Effect of aloin (0.01%) and caffeic acid (0.01%) on MMP1 protein expression [0064] The present example studies the effect of aloin (0.01%) and caffeic acid (0.01%) on MMP1 protein expression.

[0065] Figure 4 depicts the inhibition of MMP1 protein inhibition in the presence of the composition. It can be appreciated that neither the components in isolation and nor in combination of aloin (0.01%) and caffeic acid (0.01%) is able to inhibit the production of MMP1 protein production. Therefore, the composition comprising aloin and caffeic acid in a weight ratio of 1:1 is not able to exhibit anti-aging benefits.
Example 4
Effect of aloin (0.01%) and caffeic acid (0.0005%) on MMP1 protein expression
[0066] Figure 5 depicts the effect of a combination of aloin to caffeic acid in a weight ratio of 20:1.
[0067] It can be appreciated from Figure 5 that 0.01%> aloin and 0.0005%> caffeic acid in isolation are not able to inhibit MMP1 protein expression. Also, no significant reduction in the MMP1 protein expression can be observed in case of a combination of 0.01% aloin and 0.0005% caffeic acid.
[0068] Therefore, it can be inferred that the composition comprising aloin to caffeic acid at a weight ratio of 20:1 is not able to exert any significant inhibition of MMP1 protein production.
[0069] Overall, Example 2 depicts a working example which exhibits the synergistic effect of the composition comprising aloin and caffeic acid in a 10:1 weight ratio. Whereas, Example 3 (aloin to caffeic weight ratio 1:1) and Example 4 (aloin to caffeic weight ratio 20:1) depict non-working examples which fail to display any synergistic effect in inhibiting MMP1 protein expression. Therefore, it is evident that compositions comprising aloin and caffeic acid in any random ratio will not exhibit the desired effect, and significant experimentation was required to arrive at the composition of the present disclosure. Example 5 Formulation comprising the composition of the present disclosure
[0070] The composition can be used in form of a formulation that can be used for topical application in various forms of oil, serum, cream, lotion and so on.

[0072] Process for preparation of the composition comprising aloin and caffeic
acid: The composition comprising aloin and caffeic acid was prepared by obtaining
the desired weight percentage of aloin and caffeic acid, and contacting and mixing
them under ambient conditions.
[0073] Process for preparation of the formulation: The formulation as disclosed
herein was prepared by adding all the excipients and the actives (aloin and caffeic
acid) and mixing them properly under ambient conditions.
[0074] It can be contemplated that the formulation can be prepared by adding all
the components in the disclosed weight percentages. The order of adding the
components can be varied as per the process followed.
Advantages of the present disclosure
[0075] The present disclosure discloses a composition comprising aloin and caffeic acid in a specific weight ratio. The composition exhibits anti-aging effects by synergistically inhibiting the production of MMP1 protein. The actives being herbal in nature will not have side effects upon extended usage. The composition of the present disclosure can be used for dispensing in various commercially acceptable forms with ease. Also, the process for preparation is not associated with any complexities.

I/We Claim:
1. A composition comprising:
a) aloin; and
b) caffeic acid,
wherein aloin to caffeic acid has a w/w ratio in a range of 8:1 to 12:1.
2. The composition as claimed in claim 1, wherein aloin to caffeic acid has a w/w ratio of 10:1.
3. The composition as claimed in any one of the claims 1-2, wherein aloin has a weight percentage in a range of 0.005-0.015% with respect to the composition, and caffeic acid has a weight percentage in a range of 0.0005-0.002% with respect to the composition.
4. The composition as claimed in claim 3, wherein aloin has a weight percentage of 0.01% with respect to the composition, and caffeic acid has a weight percentage in a range of 0.001% with respect to the composition.
5. The composition as claimed in any one of the claims 1-4, wherein the composition further comprises at least one excipient selected from the group consisting of water, dimethyl sulfoxide (DMSO), polymer, gum base, chelating agent, emulsifier, surfactant, bulking agent, humectant, solvent, antimicrobial agent, moisturizer, fragrance, and combinations thereof.
6. A process for preparing the composition as claimed in claim 1, said process comprising:

a) obtaining aloin;
b) obtaining caffeic acid; and
c) contacting aloin with caffeic acid, to obtain the composition.
7. A process for preparing the composition as claimed in any one of the claims
1-5, said process comprising:
a) obtaining aloin;
b) obtaining caffeic acid;
c) obtaining the at least one excipient; and

d) contacting aloin, and caffeic acid with the at least one excipient, to obtain the composition.
8. The composition as claimed in any one of the claims 1-5, wherein the composition inhibits MMP1 (matrix metalloprotease 1) protein synthesis.
9. The composition as claimed in any one of the claims 1-5, wherein the composition reduces skin wrinkles caused due to aging.