FIELD OF INVENTION
[001] The present disclosure broadly relates to field of cosmetics and particularly relates to a composition comprising trehalose and oxyresveratrol for imparting anti-ageing, and skin-whitening effect.
BACKGROUND OF INVENTION
[002] In the last ten years, important progresses have been reported in the field of pharmacologic targets identification and in the development of new bioactive molecules and its combinations. These progresses were enabled, among other breakthroughs, by technological advances in genomics, proteomics and structural biology. The general practice of all these techniques together led to the identification of highly selective and potent molecules and its combination to reduce the aging process as well as treating other pathologies, such as chronic inflammatory diseases and cancer.
[003] Molecular chaperones are a fundamental group of proteins that have been involved in various cellular function. They are key components of a protein quality machinery in the cell which ensures that the folding process of any newly-synthesized polypeptide chain results in the formation of a properly folded protein and that the folded protein is maintained in an active conformation throughout its functional lifetime. Chaperones can also assist in the unfolding and degradation of misfolded proteins and in disaggregating the preformed protein aggregates. Chaperones are also involved in other cellular functions including protein translocation across membranes, vesicle fusion events, and protein secretion which essentially have role in skin pigmentation.
[004] In recent years, tremendous advances have been made in understanding of the biology, biochemistry, and biophysics of function of molecular chaperones. In addition, recent technical developments in the fields of proteomics and genomics allowed us to obtain a global view of chaperone interaction networks. There is now a growing interest in the role of molecular chaperones in skin pigmentation. Melanogenesis is regulated by a variety of signal transduction pathways. In

mammals, more than 100 genes have been shown to be involved in the process of
melanogenesis. Several of these have also been implicated in chaperones response.
[005] Skin aging is a natural process associated with a number of
pathophysiology that can reduce quality of life and longevity. Thus, various
molecular targets need to be taken into consideration for arriving at an effective
formulation for skin benefits.
[006] US5980904A discloses a skin-whitening composition comprising bearberry
extract and a reducing agent. The composition provides significant whitening of the
skin on topical application.
[007] US20120064136A1 discloses a nano-emulsion composition for
preventing/minimizing signs of ageing on the skin.
[008] Thus, there is a dire need of a composition that can effectively provide a
skin-whitening effect as well as anti-ageing effect.
SUMMARY OF THE INVENTION
[009] In an aspect of the present disclosure, there is provided a composition comprising: (a) trehalose; and (b) oxyresveratrol, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1. [0010] In an aspect of the present disclosure, there is provided a process for preparing the composition comprising: (a) trehalose; and (b) oxyresveratrol, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1, said process comprising: (i) obtaining trehalose; (ii) obtaining oxyresveratrol; and (iii) contacting trehalose and oxyresveratrol, 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 illustrates percentage of melanin inhibition in B16 mouse
melanoma cells with respect to oxyresveratrol concentrations, in accordance with an
embodiment of the present disclosure.
[0014] Figure 2 illustrates percentage of increase in activation of microtubule-
associated proteins 1A/1B light chain 3 (LC3) protein expression in B16 mouse
melanoma cells with respect to trehalose concentrations, in accordance with an
embodiment of the present disclosure.
[0015] Figure 3 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on calnexin protein expression in B16 mouse melanoma cells,
in accordance with an embodiment of the present disclosure.
[0016] Figure 4 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on calmodulin protein expression in B16 mouse melanoma
cells, in accordance with an embodiment of the present disclosure.
[0017] Figure 5 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on myosin-9 protein expression in B16 mouse melanoma cells,
in accordance with an embodiment of the present disclosure.
[0018] Figure 6 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on heat shock protein 70 ((HSP 70) expression in B16 mouse
melanoma cells, in accordance with an embodiment of the present disclosure.
[0019] Figure 7 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on heat shock protein 90 (HSP 90) expression in B16 mouse
melanoma cells, in accordance with an embodiment of the present disclosure.
[0020] Figure 8 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on profilin-1 protein expression in B16 mouse melanoma cells,
in accordance with an embodiment of the present disclosure.

[0021] Figure 9 illustrates the effect of a combination of lOOmM trehalose and
lOuM oxyresveratrol on cyclophilin-A in B16 mouse melanoma cells, in accordance
with an embodiment of the present disclosure.
[0022] Figure 10 illustrates the effect of individual actives and various combinations
on calnexin protein expression in B16 mouse melanoma cells, in accordance with an
embodiment of the present disclosure.
[0023] Figure 11 illustrates the effect of individual actives and various combinations
on calmodulin protein expression in B16 mouse melanoma cells, in accordance with
an embodiment of the present disclosure.
[0024] Figure 12 illustrates a schematic representation of combinatory effect of
oxyresveratrol and trehalose on skin health, in accordance with an embodiment of
the present disclosure.
[0025] Figure 13 illustrates a schematic representation of the role of calnexin,
calmodulin, HSP70, HSP90 a, and myosin-9 in inducing melanogenesis process, in
accordance with an embodiment of the present disclosure.
[0026] Figure 14 illustrates a schematic representation of the role of profilin-1 and
cyclophilin-A in reducing skin aging process, in accordance with an embodiment of
the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0027] 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
[0028] 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.
[0029] 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.
[0030] 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".
[0031] 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.
[0032] The term "including" is used to mean "including but not limited to".
"Including" and "including but not limited to" are used interchangeably.
[0033] 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.
[0034] Oxyresveratrol (2, 4, 3, 5-tetrahydroxystilbene) is a derivative of
resveratrol. It's linking C = C double bond has a trans conformation and allows the
formation of a conjugated system throughout the molecule. Oxyresveratrol is a
yellow solid with a melting-point of 199-204°C (>98%), a molecular formula of
C14H12O4 and a molecular weight of 244.24 MW. Oxyresveratrol is a hydroxyl-
substituted stilbene found in the roots, leaves, stem and fruit of many widely
distributed plants. Oxyresveratrol has various kinds of biological activities
including antineoplastic activity, antioxidant activity, tyrosinase inhibitory activity
and ability of boosting the immune system. Oxyresveratrol is a potential substance
to be utilized to produce medicines, cosmetics and health supplements.

[0035] Trehalose is a non-reducing sugar with similar chemical structure and characteristics to those of sucrose. Trehalose, a naturally occurring nontoxic disaccharide found in plants, insects, microorganisms and invertebrates, but not in mammals, was reported to function as a mechanistic target of the rapamycin (mTOR)-independent inducer of autophagy. It is suggested that this sugar is highly connected with life; it supposedly protects life from damages by freezing or drying and plays an important role in the development of tolerance. Trehalose possesses antioxidant and anti-inflammatory properties that protects against aging in lower organisms. Trehalose also improves physiological function in a number of pro-oxidant and inflammatory age-associated diseases. The mechanisms by which trehalose protects against oxidative stress and inflammation are incompletely understood but may be mediated through the ability of trehalose to up-regulate autophagy. Importantly from a therapeutic standpoint, trehalose has been investigated extensively for safety, toxicity and carcinogenesis because of its widespread use in foods, cosmetics, and pharmaceuticals. Furthermore, trehalose received the Food and Drug Administration's designation "generally recognized as safe" in 2000. Together, the safety profile, low cost, and high availability to trehalose make it a feasible and attractive agent for clinical use. [0036] 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.
[0037] For the purposes of the present disclosure, the composition of trehalose and oxyresveratrol which comprises the formulation of the present disclosure is only the partial solution for skin anti-ageing mechanism.
[0038] To address the problem of skin ageing and hyper pigmentation, an effective solution would be to intervene the process of ageing and pigmentation at the molecular level. As discussed, one attractive target is chaperone system. The goal of this disclosure is to offer anew composition to promote skin depigmentation and anti-aging process in skin cells and the same composition can also be used to promote anti-melanogenesis activity. The basis for the present disclosure is that

skin depigmentation results from the dysregulation of key regulatory proteins (i.e., protein signatures) which in turn, interacting and overlapping networks and intracellular pathways. Dysregulation of protein functions leads to physiological aging of tissues, cells and subcellular components. The present study provides information that will guide future research and suggest new therapies for preventing, mitigating or treating skin pathologies related to aging. [0039] The present disclosure discloses a composition comprising trehalose and oxyresveratrol and its effect in modulating a variety of targets. The targets studied have been represented in the following paragraphs:
1. Calnexin is an abundant 90kDa chaperone protein that resides in the membrane
of the endoplasmic reticulum. It is a calcium-binding protein that interacts with
newly synthesized glycoproteins in the endoplasmic reticulum. It may act in
assisting protein assembly and/or in the retention within the ER of unassembled
protein subunits. Calnexin (CNX) is a lectin chaperones, components of the
endoplasmic reticulum quality control system (ERQC) that associate with proteins
during folding Formation of a fully functional, active tyrosinase requires that the
tyrosinase translated from tyrosinase mRNA on ribosomes becomes translocated
into the ER for folding and further maturation. Tyrosinase folds through several
inactive intermediates, at least two of which are recognized by the ER chaperone,
calnexin. If the association with calnexin is prevented, more rapid folding occurs,
the resulting protein fails to bind copper and is inactive. If dissociation from
calnexin is inhibited, folding is prevented; the protein does not go through the
normal secretory pathway and is targeted for degradation. Hence, calnexin has an
important role in skin pigmentation process.
2. Calmodulin (CaM) (an abbreviation for calcium-modulated protein) is a
multifunctional intermediate calcium-binding messenger protein expressed in all
eukaryotic cells. It is an intracellular target of the secondary messenger Ca2+, and
the binding of Ca2+ is required for the activation of calmodulin. Calcium and
calmodulin are intimately involved in both the production and the degradation of
cyclic AMP. The majority of the actions of calcium within the cell are now known
to be mediated by its binding to the ubiquitous calcium binding protein calmodulin.

It is the calcium-calmodulin complex which then interacts with intracellular proteins, usually enzymes, to affect the biological response such as skin pigmentation.
3. Hsp70 family proteins generally function as molecular chaperones for the post-translational modification of target proteins and to prevent aggregate formation, promoting maintenance of cellular homeostasis following cellular stresses. In addition to promoting optimal protein folding, Hsp70 proteins also function in a variety of cellular biological processes such as melanogenesis. Two-dimensional gel electrophoresis (2-DGE) and mass spectrometry demonstrated that heat shock protein 70-1A (Hsp70-1A) protein levels were significantly higher in AA melanocytes compared to white melanocytes. Hsp70-1A expression significantly correlated with levels of tyrosinase, the rate-limiting melanogenic enzyme, consistent with a proposed role for Hsp70-family members in tyrosinase post-translational modification. Additionally, pharmacologic inhibition and siRNA-mediated down-regulation of Hsp70-1A correlated with pigmentation changes in cultured melanocytes, modified human skin substitutes and ex vivo skin.
4. Melanogenesis is a complex metabolic pathway controlled by a family of enzymes known as tyrosinase-related proteins, which are synthesized and glycosylated in the ER and Golgi and then further trafficked to melanosomes where they synthesize and deposit melanin. HSP90 family, also known as HSP90 is a molecular chaperone localized in the lumen of the endoplasmic reticulum (ER) and is important for protein maturation and homeostasis. A proteomics study from other group showed that the chaperone hsp90 is also present within melanosome and studies revealed that hsp90 positively regulates melanogenesis and melanosome development by facilitating tyrosinase trafficking and enzymatic function via chaperoning the Wnt canonical pathway.
5. Profilin-1 as an actin-binding protein is a class of small molecule proteins (12 to 15 KD) and is widely distributed in various types of cells with highly conserved sequences. It plays an essential role in the regulation of cytoskeleton by promoting actin polymerization and remodeling. PFN-1 plays an important role in AGEs-induced endothelial abnormalities via promoting cytoskeleton rearrangement and

redistribution and silencing the expression of PFN-1 attenuated AGEs-induced myocardium injury including cardiac hypertrophy and fibrosis in vivo. The reorganization and redistribution of the cytoskeleton regulated by profilin-1 mediates endothelial cell contraction, which results in vascular hyperpermeability. Recently, it has been reported that overexpression of profilin-1 up-regulated the expression of ICAM-1, increased endothelial cell permeability, induced endothelial cell apoptosis and promoted endothelial cell migration, adhesion and focal contact formation and silencing profilin-1 gene expression provided significant protection on endothelial cells. Profilin-1 might act as an ultimate and common cellular effector in the process of metabolic memory (endothelial abnormalities) mediated by advanced glycation end products via the ROS/PKC or ROS/NF-K;B signaling pathways.
6. Peptidylprolyl isomerase A (PPIA). also known as cyclophilin A (CypA) or rotamase A is an enzyme that in humans is encoded by the PPIA gene on chromosome 7. As a member of the peptidyl-prolyl cis-trans isomerase (PPIase) family, this protein catalyzes the cis-trans isomerization of proline imidic peptide bonds, which allows it to regulate many biological processes, including intracellular signaling, transcription, inflammation, and apoptosis. Moreover, the enzyme participates in inflammatory and apoptotic processes in extracellular settings. In the presence of reactive oxygen species (ROS), vascular smooth muscle cells (VSMCs), monocytes/macrophages, and endothelial cells (ECs) secrete PPIA to induce an inflammatory response and mitigate tissue injury. Another interesting aspect of CyPA functions is its correlation with aging. Recently, Li et al. showed that the expression of CyPA increased with aging in normal skin, regardless of whether it was exposed or shaded. Similarly, it has been discovered that the expression level of CyPA is upregulated in the epidermis of aged people compared with younger people. Further, CyPA expression was found significantly higher in old rats than in the younger ones. It is believed that the development of agents that selectively inactivate CyPA or that block binding of CyPA to its receptor could prove to be a fruitful approach to forestalling several human diseases.

[0040] The present disclosure provides a composition comprising trehalose and oxyresveratrol, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10000:1. The composition comprises trehalose having a weight percentage in a range of 2.5-4.5% with respect to the composition, and oxyresveratrol having a weight percentage in a range of 0.0002-0.0003% with respect to the composition. The effect of the composition has been studied in B-16 mouse melanoma cells. As a part of the present disclosure, the composition is shown to synergistically down-regulate the expression of: (i) melanogenesis promoting proteins such as calnexin, calmodulin, HSP 70, HSP 90 a, and myosin-9; (ii) skin-ageing promoting proteins such as profilin-1 and cyclophilin-A. Also provided is a method to prepare the composition as disclosed herein. [0041] 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. [0042] 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. [0043] In an embodiment of the present disclosure, there is provided a composition comprising: (a) trehalose; and (b) oxyresveratrol, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1. In another embodiment of the present disclosure, trehalose to oxyresveratrol weight ratio is in a range of 9800:1 to 10200:1. In yet another embodiment of the present disclosure, trehalose to oxyresveratrol weight ratio is in a range of 9900:1 to 10100:1. [0044] In an embodiment of the present disclosure, there is provided a composition comprising: (a) trehalose; and (b) oxyresveratrol, wherein trehalose to oxyresveratrol weight ratio is 10000:1.

[0045] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) trehalose having a concentration in a range of 75mM tol25mM;
and (b) oxyresveratrol having a concentration in a range of 7.5uM to 15uM,
wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1.
In another embodiment of the present disclosure, trehalose is having a
concentration in a range of 90mM to llOmM, and oxyresveratrol is having a
concentration in a range of 9uM to 12uM.
[0046] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) trehalose having a concentration of lOOmM; and (b) oxyresveratrol
having a concentration of lOuM, wherein trehalose to oxyresveratrol weight ratio is
in a range of 9500:1 to 10500:1.
[0047] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) trehalose having a concentration of lOOmM; and (b) oxyresveratrol
having a concentration of lOuM, wherein trehalose to oxyresveratrol weight ratio is
10000:1.
[0048] In an embodiment of the present disclosure, there is provided a composition
comprising: (a) trehalose having a weight percentage in a range of 2.5-4.5% with
respect to the composition; and (b) oxyresveratrol having a weight percentage in a
range of 0.0002-0.0003% with respect to the composition, wherein trehalose to
oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1.
[0049] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein the composition further comprises at least one
excipient selected from the group consisting of polymer, gum base, chelating agent,
emulsifier, surfactant, humectant, solvent, antimicrobial agent, moisturizer,
fragrance, bulking agent, water, dimethyl sulfoxide (DMSO), and combinations
thereof.
[0050] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein the composition further comprises at least one
excipient selected from the group consisting of polymer, gum base, chelating agent,
emulsifier, surfactant, humectant, solvent, antimicrobial agent, moisturizer,
fragrance, bulking agent, water, and combinations thereof, and wherein the at least

one excipient is selected from the group consisting of: (a) polymers having a weight percentage in the range of 0.2 - 0.5% with respect to the composition; (b) gum base having a weight percentage in the range of 0.1 - 0.5% with respect to the composition; (c) chelating agents having a weight percentage in the range of 0.05 -0.2% with respect to the composition; (d) emulsifiers having a weight percentage in the range of 1 - 5% with respect to the composition; (e) surfactants having a weight percentage in the range of 0.5 - 3% with respect to the composition; (f) humectants having a weight percentage in the range of 0.1 - 1% with respect to the composition; (g) solvents having a weight percentage in the range of 1 - 3% with respect to the composition; (h) antimicrobial agents having a weight percentage in the range of 0.1 - 1% with respect to the composition; (i) moisturizers having a weight percentage in the range of 2 - 3% with respect to the composition; (j) bulking agent having a weight percentage in the range of 0.5-5% with respect to the composition.
[0051] In an embodiment of the present disclosure, there is provided a composition comprising: (a) trehalose having a weight percentage in a range of 2.5-4.5% with respect to the composition; (b) oxyresveratrol having a weight percentage in a range of 0.0002-0.0003% 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 trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1.
[0052] In an embodiment of the present disclosure, there is provided a composition comprising: (a) trehalose having a weight percentage in a range of 2.5-4.5% with respect to the composition; (b) oxyresveratrol having a weight percentage in a range of 0.0002-0.0003% 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 acryloyldimethyl 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 trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1.
[0053] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) trehalose; and (b) oxyresveratrol, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1, said process comprising: (i) obtaining trehalose; (ii) obtaining oxyresveratrol; and (iii) contacting trehalose and oxyresveratrol, to obtain the composition. [0054] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) trehalose having a weight percentage in a range of 2.5% to 4.5% with respect to the composition; and (b) oxyresveratrol having a weight percentage in a range of 0.0002% to 0.0003% with respect to the composition, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1, said process comprising: (i) obtaining trehalose; (ii) obtaining oxyresveratrol; and (iii) contacting trehalose and oxyresveratrol, to obtain the composition.
[0055] In an embodiment of the present disclosure, there is provided a process for preparing the composition comprising: (a) trehalose having a concentration in a range of 75mM to 125mM with respect to the composition; and (b) oxyresveratrol having a concentration in a range of 7.5uM to 15uM with respect to the composition, wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1, said process comprising: (i) obtaining trehalose; (ii) obtaining

oxyresveratrol; and (iii) contacting trehalose and oxyresveratrol, to obtain the
composition.
[0056] In an embodiment of the present disclosure, there is provided a process for
preparing the composition comprising: (a) trehalose; (b) oxyresveratrol; and (c) at
least one excipient, wherein trehalose to oxyresveratrol weight ratio is in a range of
9500:1 to 10500:1, said process comprising: (i) obtaining trehalose; (ii) obtaining
oxyresveratrol; (iii) obtaining the at least one excipient; and (iv) contacting
trehalose, and oxyresveratrol with the at least one excipient, to obtain the
composition.
[0057] In an embodiment of the present disclosure, there is provided a process for
preparing the composition comprising: (a) trehalose having a weight percentage in
a range of 2.5% to 4.5% with respect to the composition; (b) oxyresveratrol having
a weight percentage in a range of 0.0002% to 0.0003% with respect to the
composition; and (c) at least one excipient, wherein trehalose to oxyresveratrol
weight ratio is in a range of 9500:1 to 10500:1, said process comprising: (i)
obtaining trehalose; (ii) obtaining oxyresveratrol; (iii) obtaining the at least one
excipient; and (iv) contacting trehalose, and oxyresveratrol with the at least one
excipient, to obtain the composition.
[0058] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein the composition exhibits anti-ageing effects.
[0059] embodiment of the present disclosure, there is provided a composition as
described herein, wherein the composition exhibits skin-whitening effects.
[0060] In an embodiment of the present disclosure, there is provided a formulation
as described herein, wherein said composition can be for topical application.
[0061] In an embodiment of the present disclosure, there is provided a formulation
as described herein, wherein said composition can be in the form of an aerosol.
[0062] In an embodiment of the present disclosure, there is provided a formulation
as described herein, wherein said composition can be in the form of a lotion.
[0063] In an embodiment of the present disclosure, there is provided a formulation
as described herein, wherein said composition can be in form of an oil.

[0064] In an embodiment of the present disclosure, there is provided a formulation as described herein, wherein said composition can be in the form of a serum. [0065] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[0066] 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.
[0067] The working and non-working examples of the composition comprising
trehalose and oxyresveratrol will provide support to the weight percentage range
and weight ratio as claimed in the present disclosure. The effect of the present
composition on various molecular targets will be depicted in the forthcoming
sections.
Material and Methods
[0068] Oxyresveratrol was procured from Sami Labs., India. Stock solution was
prepared in DMSO. Trehalose was purchased from Sigma. Stock solution was
prepared in water.
[0069] Mouse melanoma cells (B16) and HaCat cells were procured from ATCC,
India.
[0070] DMEM media was purchased from Gibco, USA.
Protocol of experiments performed

Proteomics study; The protocol was spread over a period of 5 days:
[0071] Day 1: B-16 cells were seeded in T 25- flask and incubated in 5% C02 incubator at 37 °C.
[0072] Day 2: B16 cells were exposed to 10 uM oxyresveratrol and 100 millimolar of trehalose and combination of oxyresveratrol (10 uM) + trehalose (100 mM) for 72 hours.
[0073] Day 5: Cells were lysed and processed for proteomics analysis as per the following protocol
Proteomic analysis; 30 ug of total protein of each sample (untreated B16 cell lines treated with oxyresveratrol (10 uM), trehalose (100 mM) and its combination (10 uM) of oxyresveratrol and trehalose (100 mM) was subjected to in-solution digestion method described later. The peptide mixtures were separated using a reverse phase column nano-high performance liquid chromatography (HPLC) using pico-frit columns (Thermo) (0.075 mm x 150 mm, CI8). The peptides were eluted using a gradient 5 to 90 % of acetonitrile (95%) at a flow rate of 350 nl/min for 60 min. A linear trap quadrapole (LTQ) orbitrap velos mass spectrometer equipped with nano-electron source ionization (ESI) source was used to acquire spectra. A 75-um metal emitter spray tip was used and the spray voltage was set at 2 kV. The instrument was set under stable spray and conditioned before loading the samples. The instrument was operated in data-dependent mode with dynamic exclusion enabled. The tandem mass spectrometry (MS/MS) spectra on the top 20 most abundant peptide ions in full MS scan were obtained. The normalized collision-induced dissociation (CID) was set at 35% for MS/MS. All MS/MS spectra were searched against the human protein database from uniprot using SEQUEST algorithm incorporated in proteome discoverer 1.4. (Using trypsin as cleavage enzyme).
In solution digestion method; In-solution protein digestion, is a proteomics protocol in which proteins are digested in solution. 100 ug of cell protein extract was resuspended in 47 ul of 6 M urea, 50 mM ammonium bicarbonate solution.

One ul of 250 mM TCEP (tris (2 carboxyethyl) phosphine) was added in solution and the sample was incubated at 37°C for 30 minutes. Further 2 ul of 625 mM iodoacetamide was added and the sample was incubated in dark at room temperature (25±2°C) for 45 minutes. Finally, in-solution digestion was carried by addition of 450 ul trypsin in 50 mM ammonium bicarbonate with protein digest ratio of 50:1. The digested peptides were purified with OASIS 1 ml Cartridge system form Waters Scientific (WAT094225).
Nano-HPLC & Mass Spectrometry; The purified / enriched samples were dried and re-suspended in 0.1% formic acid with 5% acetonitrile in MS water. The peptides were loaded onto an EASY-nano LC system (Proxeon). Peptide mixtures were separated on a CI8 reverse phase column (PicoFrit capillary column, 75 urn* 10cm, New Objective) using a linear gradient of solvent A (0.1% formic acid in 5% acetonitrile) and Solvent B (0.1% formic acid in 95%acetonitrile) at a flow rate 250 nL/min directly into a LTQ-OrbitrapVelos mass spectrometer (Thermo Scientific, San Jose, CA, USA) with nano-ESI source with spray voltage was set at 1.6 KV. Stable spray condition maintained prior to sample loading. The instrument was operated in data-dependent mode with dynamic exclusion enabled. The MS/MS spectra on the top 20 most abundant peptide ions in full MS scan were obtained. The normalized CID was set at 35% for MS/MS.
Data Analysis; Data analysis pipeline were set to carry out the analysis of data produced by Proteome discoverer software version 1.4. An in-house Python based tool was used for identifying the differentially expressed proteins across and among the samples. The data were searched against the mouse protein database of Uniprot. The search was performed choosing trypsin as specific enzyme. A maximum of one missed cleavage was allowed. Peptide (parent ion) tolerance of 2.5 Da and fragment ion tolerance of 1 Da were allowed, Oxidation (M) as variable modifications and fixed modification of carbamidomethylation (C) were used. The combinational studies using oxyresveratrol and trehalose on mouse melanoma cells (B16 Cells) were performed using in-solution LC MS/MS approach. A total of

3306 unique proteins were identified and 945 unique proteins were identified in the basal group. 983 unique proteins were identified in the cells treated with oxyresveratrol at 10 micro molar (10 uM). 1048 unique proteins were identified in the 100 milli molar ( mM) trehalose treated cells. 688 unique proteins were identified in cells treated with combination of molecules comprising of oxyresveratrol at 10 micro molar + trehalose at 100 milli molar (mM).
Quantitative proteomic data analysis was performed after global data normalization. All the above proteins are deregulated in the various physiological conditions directly or indirectly and the above combination synergistically modulated the protein levels in a favorable way.
[0074] Cell culture conditions; B 16 cells obtained from ATCC were used for this study. B 16 cells will be seeded (at a density of 104 cells/cm2) in appropriate culture medium (DMEM and 10 % FBS). Cells will be treated with 10 nm of Forskolin before treating with test components. Cell Culture vessels were incubated at 37 °C with a humidified 5 % CO2 atmosphere. Cell growth and morphology was monitored on day 5, 7 and 9 post-seeding. After 3 days of the treatment of test compounds (Non-cytotoxic concentrations), the B 16 cells were collected for the total melanin assay. The assay was performed as mentioned in the below protocol. [0075] Melanin determination; The following steps were followed for determining melanin content in B 16 cells.
1. B16 cells (approximately 105 cells) were pelleted in eppendorf tube.
2. The cells were solubilized in 1 ml of IN NaOH / 10% DMSO for 2 hours at 80°C
3. Similarly, the tubes were prepared for standard curve using synthetic melanin (Sigma) covering concentration range of 0-20 (ig/ml.
4. The tubes were centrifuged at 12000 g for 10 min at RT, and supernatants were transferred to fresh tubes.
5. The absorbance of the supernatants was measured at 405 nm. The melanin content was further determined using standard curve generated for synthetic

melanin. Melanin contents was determined by the absorbance/ug of protein in a
cell extract.
The melanin content is expressed percentage of inhibition when compared to
control
Determination of LC3 content; Autophagy was evaluated by using western blot to detect the autophagic markers - microtubule associated protein light chain 3 (LC3) levels in cell culture supernatants were measured by a commercial enzyme-linked immunosorbent assay kit (ELISA, LifeSpan BioSciences, Katy, USA) following the manufacturer's protocol.
The reagents and samples were brought to room temperature without additional heating and mixed thoroughly by gently swirling before pipetting (avoid foaming). All the reagents, working standards, and samples were prepared as directed in the manual.
1. 100 ul of standard, blank, or sample per well were added, covered with a plate sealer, and incubated for 2 hours at 37°C.
2. The liquid of each well was aspirated.
3. 100 ul of detection reagent A working solution to each well were added, covered with a plate sealer, and gently agitated to ensure thorough mixing. Incubated for 1 hour at 37°C.
4. The liquid was aspirated from each well and washed 3 times. Washed by adding approximately 350 ul of wash buffer using a squirt bottle, multi-channel pipette. Allowed each wash to sit for 1-2 minutes before completely aspirating. After the last wash, aspirated to remove any remaining wash buffer then invert the plate and tap against clean absorbent paper.
5. 100 ul of detection reagent B working solution to each well were added, covered with a new plate sealer, and incubated for 30-60 minutes at 37°C.
6. Aspirated the liquid from each well and washed 5 times as outlined in step 4.

7. Added 90 ul of TMB Substrate solution to each well, covered with a new plate sealer, and incubated for 15-30 minutes at 37°C. Protected from light and monitor periodically until optimal color development has been achieved.
8. Added 50 ul of stop solution to each well. The blue color was changed to yellow immediately. The Stop Solution were added to wells in the same order and timing as the TMB substrate solution.
9. Determined the optical density (OD value) of each well immediately using a microplate reader set to 450 nm.
Example 1
Effect of oxyresveratrol on melanin content
[0076] The present example depicts the effect of oxyresveratrol on melanin content in B16 cells. Different concentrations of oxyresveratrol such as 2uM, 4uM, 6uM, 8uM, lOuM, 12uM, and 14uM were tested on B16 cells.
[0077] Referring to Figure 1, it can be appreciated that a steady increase in the rate of melanin inhibition can be observed till lOuM (42.47% inhibition). At higher concentrations, very slight increase in melanin inhibition can be observed. Therefore, to avoid unnecessary toxicity because of high oxyresveratrol content, lOuM was used for preparing a combinatorial composition.
Example 2
Effect of trehalose on LC3 protein expression
[0078] The present example depicts the effect of trehalose on percentage of increase in activation of microtubule-associated proteins 1A/1B light chain 3 (LC3) protein expression in B16 mouse melanoma cells. Trehalose concentrations in a range of lOmM to 200mM was tested for the effect on LC3 protein expression in B16 cells. LC3 is an autophagy marker and was observed that trehalose induces the autophagy in cells.

[0079] Referring to Figure 2, it can be observed that the percentage expression of LC3 gradually increases with increase in trehalose concentration. But the percentage of LC3 expression in case of lOOmM and 200mM trehalose is similar. Thus, lOOmM trehalose was used for making synergistic combination with oxyresveratrol.
Example 3
Effect of a combination of trehalose and oxyresveratrol on calnexin protein
expression
[0080] The present example depicts the effect of the present composition on calnexin protein expression using a method as described previously. [0081] Referring to Figure 3, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol in isolation no significant change in the expression can be observed. In case of treatment with the composition of the present disclosure, a significant fold change of 0.88-fold can be observed. Thus, it can be appreciated that a synergistic decrease in expression of calnexin can be observed in presence of the present composition.
[0082] Calnexin is a calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins. Calnexin, which could be a candidate as a chaperone for sorting and maturation of tyrosinase family glycoproteins in melanogenesis cascade. Therefore, by down regulating the expression of calnexin, the present composition affects the melanogenesis cascade, thereby exhibiting skin-whitening effect.
Example 4
Effect of a combination of trehalose and oxyresveratrol on calmodulin
expression
[0083] The present example depicts the effect of the present composition on calmodulin expression using a method as described previously.

[0084] Referring to Figure 4, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol in isolation no significant change in the expression can be observed. In case of treatment with the composition of the present disclosure, a significant fold decrease of 6.69-fold can be observed. Thus, it can be appreciated that a synergistic decrease in expression of calmodulin can be observed in presence of the present composition.
[0085] Calmodulin is a calcium modulated protein. It is abundant in the cytoplasm of all higher cells and has been highly conserved through evolution. Calmodulin acts as an intermediary protein that senses calcium levels and relays signals to various calcium-sensitive enzymes, ion channels and other proteins. Calmodulin is a small dumbbell-shaped protein composed of two globular domains connected together by a flexible linker. This protein plays a crucial role in promoting melanogenesis cascade, thereby down-regulating calmodulin exhibits a reduction in skin pigmentation.
Example 5
Effect of a combination of trehalose and oxyresveratrol on Myosin-9 protein
expression
[0086] The present example depicts the effect of the present composition on Myosin-9 protein expression using a method as described previously. [0087] Referring to Figure 5, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol in isolation no significant change in the expression can be observed. In case of treatment with the composition of the present disclosure, a significant fold decrease of 3.25-fold can be observed. Thus, it can be appreciated that a synergistic decrease in expression of Myosin-9 protein can be observed in presence of the present composition.
[0088] Myosin-9 protein function in Ca2+/calmodulin signaling and are also found in a number of other types of protein, for example voltage operated channels, phosphatases and Ras GTPase-activating protein. Myosin-9 play an important role in mechanism that controls melanin transfer from human cutaneous melanocytes to human cutaneous keratinocytes. Considering the crucial role played by Myosin-9

protein, the down regulation of the protein by the present composition is a promising feature of displaying skin-whitening effect. Example 6
Effect of a combination of trehalose and oxyresveratrol on Heat shock 70 (HSP 70) protein expression
[0089] The present example depicts the effect of the present composition on HSP 70 protein expression using a method as described previously. [0090] Referring to Figure 6, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol a slight increase in the expression of HSP 70 can be observed. In case of treatment with the composition of the present disclosure, surprisingly, a significant fold decrease of 1.83-folds can be observed. Thus, it can be appreciated that a synergistic decrease in expression of HSP 70 protein can be observed in presence of the present composition. [0091] Heat shock 70 family chaperone mtHsp70/mortalin (Wadhwa et al. 1993) are critically involved in the process of melanogenesis. Two-dimensional gel electrophoresis (2-DGE) and mass spectrometry demonstrated that Heat Shock Protein 70-1A (Hsp70-1A) protein levels were significantly higher in African American; melanocytes compared to White melanocytes. Hsp70-1A expression significantly correlated with levels of tyrosinase, the rate-limiting melanogenic enzyme, consistent with a proposed role for Hsp70-family members in tyrosinase post-translational modification. Epidermal Hsp70 contributes to the diversity of skin color by regulating the amount of melanin synthesized in melanocytes and modulating autophagic melanosome degradation in keratinocytes. Therefore, the present composition down regulates a crucial target thereby affecting the melanogenesis cascade.
Example 7
Effect of a combination of trehalose and oxyresveratrol on HSP 90 protein expression
[0092] The present example depicts the effect of the present composition on HSP 90 protein expression using a method as described previously.

[0093] Referring to Figure 7, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol in isolation no significant change in the expression can be observed. In case of treatment with the composition of the present disclosure, a significant fold change of 1.74-fold can be observed. Thus, it can be appreciated that a synergistic decrease in expression of HSP 90 protein can be observed in presence of the present composition.
[0094] HSP 90 is a molecular chaperone that promotes the maturation, structural maintenance and proper regulation of specific target proteins involved for instance in cell cycle control and signal transduction. Hsp 90 is a constitutively expressed heat shock protein in the endoplasmic reticulum whose expression is further upregulated upon ultraviolet irradiation. It is involved in blocking late melanosome maturation and also involved in melanosome translocation
Example 8
Effect of a combination of trehalose and oxyresveratrol on Profilin-1 protein expression
[0095] The present example depicts the effect of the present composition on Profilin-1 protein expression using a method as described previously. [0096] Referring to Figure 8, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol in isolation no significant change in the expression can be observed. In case of treatment with the composition of the present disclosure, a significant fold change of 1.14-fold can be observed. Thus, it can be appreciated that a synergistic decrease in expression of Profilin-1 protein can be observed in presence of the present composition.
[0097] Profilins are small proteins involved in actin dynamics. In accordance with this function, they are found in all eukaryotes and are structurally highly conserved. Protein encoded by this gene is a ubiquitous actin monomer-binding protein belonging to the profilin family. Profilin 1 in regulating the cytoskeletal architecture and dynamics, maintaining cell structure integrity. The protein thus contributes to ageing by participating in inflammation mediated responses. Therefore, a synergistic inhibition of Profilin-1 thereby exhibits anti-ageing effect.

Example 9
Effect of a combination of trehalose and oxyresveratrol on Cyclophilin A (CyPA) protein expression
[0098] The present example depicts the effect of the present composition on Cyclophilin A protein expression using a method as described previously. [0099] Referring to Figure 9, it can be appreciated that in case of treatment with lOOmM trehalose and lOuM oxyresveratrol in isolation no significant change in the expression can be observed. In case of treatment with the composition of the present disclosure, a significant down regulation of 1.36-fold can be observed. Thus, it can be appreciated that a synergistic decrease in expression of Cyclophilin A protein can be observed in presence of the present composition. [00100] Cyclophilin A (CyPA) is a ubiquitously distributed protein belonging to the immunophilin family. CyPA has peptidyl prolyl cis-trans isomerase (PPIase) activity, which regulates protein folding and trafficking. Although CyPA was initially believed to function primarily as an intracellular protein, recent studies have revealed that it can be secreted by cells in response to inflammatory stimuli. As CyPA promotes skin ageing by mediating inflammation, therefore, the present composition exerts anti-ageing effect by synergistically inhibiting CyPA expression.
Example 10
Effect of different actives in isolation, and different combinations on calnexin protein expression
[00101] The present example is a non-working example in which effect of different actives (trehalose, oxyresveratrol, and dihydro-oxyresveratrol) and also effect of different compositions are depicted. Effect of the following actives and combination were tested: (i) trehalose lOOmM; (ii) oxyresveratrol lOuM; (iii) dihydro-oxyresveratrol lOuM; (iv) a combination of lOuM oxyresveratrol and lOuM dihydro-oxyresveratrol; (v) a combination of lOuM oxyresveratrol and

lOOmM trehalose; and (vi) a combination of lOuM oxyresveratrol, lOuM oxyresveratrol, and 1 OOmM trehalose
[00102] Referring to Figure 10, the combination of lOuM oxyresveratrol, and 100 mM trehalose is the composition as disclosed in the present disclosure. It can be appreciated that only the composition of the present disclosure is able to down regulate the expression of calnexin in a synergistic manner. The combination comprising all the three actives is also not able to exhibit any synergistic down regulation effect.
Example 11
Effect of different actives in isolation, and different combinations on calmodulin protein expression
[00103] The present example is a non-working example in which effect of different actives (trehalose, oxyresveratrol, and dihydro-oxyresveratrol) and also effect of different compositions are depicted. Effect of the following actives and combination were tested: (i) trehalose lOOmM; (ii) oxyresveratrol lOuM; (iii) dihydro-oxyresveratrol lOuM; (iv) a combination of lOuM oxyresveratrol and lOuM dihydro-oxyresveratrol; (v) a combination of lOuM oxyresveratrol and lOOmM trehalose; and (vi) a combination of lOuM oxyresveratrol, lOuM oxyresveratrol, and 1 OOmM trehalose
[00104] Referring to Figure 11, the combination of lOuM oxyresveratrol, and 100 mM trehalose is the composition as disclosed in the present disclosure. It can be appreciated that only the composition of the present disclosure is able to down regulate the expression of calmodulin in a synergistic manner. The combination comprising all the three actives is also not able to exhibit any synergistic down regulation effect. Thus, it can be said that any random combination of two or three actives does not produce the desired result, Therefore, considerable experimentation was needed to arrive at the composition as disclosed in the present disclosure.
[00105] The combined effect of oxyresveratrol and trehalose has been depicted as a scheme and is represented by Scheme-1, depicted by Figure 12.

The Scheme-1 (Figure 12) along with different examples clearly depict combinatory effect of oxyresveratrol and trehalose on skin health with respect to modulating the molecular chaperones to enhance the aging associated skin depigmentation. The composition targets chaperones to address the problems of skin ageing and skin hyperpigmentation.
[00106] Scheme-2 (Figure 13) presents the role of calnexin, calmodulin, HSP70, HSP90 a, and Myosin-9 in inducing melanogenesis process (skin pigmentation), and the inhibitory effect of oxyresveratrol and trehalose combination.
[00107] Scheme 3 (Figure 14) depicts the role of Profilin-1 and Cyclophilin-A in
reducing skin aging process and the modulating effect of oxyresveratrol and
trehalose combination.
Example 19
Formulation comprising the composition of the present disclosure
[00108] 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.
[00109] Table 1 depicts a formulation depicting the composition of present
disclosure.
Table 1: Formulation table
Constituents % w/w
Pasteurized (distilled) water* 82.77
ammonium acryloyl - dimethyltaurate / VP 0.05
copolymer
acrylates/C 10-30 alkyl acrylate crosspolymer 0.17
xanthangum 0.10
disodium EDTA OH)
magnesium sulfate 0.70


components.
[00110] Process for preparation of the formulation: The formulation as disclosed herein was prepared by adding all the excipients and the actives (trehalose and 5 oxyresveratrol) and mixing them properly under ambient conditions.
[00111] 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.
10 Advantages of the present disclosure

[00112] The present disclosure discloses a composition comprising trehalose and oxyresveratrol in a specific weight ratio. The composition exhibits various skin benefits primarily by targeting molecular chaperones. The composition exhibits anti-ageing effect, and skin-whitening effect by modulating various molecular targets. One of the significant advantage of the present disclosure being the fact that the composition is able to target multiple integrated pathways thereby providing skin beneficial effects. The composition provides a solution at molecular levels thereby providing effective and long-lasting benefits upon its usage.

I/We Claim;
1. A composition comprising:
a) trehalose; and
b) oxyresveratrol,
wherein trehalose to oxyresveratrol weight ratio is in a range of 9500:1 to 10500:1.
2. The composition as claimed in claim 1, wherein trehalose to oxyresveratrol weight ratio is 10000:1.
3. The composition as claimed in any one of the claims 1, or 2, wherein trehalose has a weight percentage in a range of 2.5-4.5% with respect to the composition, and oxyresveratrol has a weight percentage in a range of 0.0002-0.0003% with respect to the composition.
4. The composition as claimed in any one of the claims 1 to 3, wherein the composition further comprises at least one excipient selected from the group consisting of polymer, gum base, chelating agent, emulsifier, surfactant, humectant, solvent, antimicrobial agent, moisturizer, fragrance, bulking agent, water, dimethyl sulfoxide (DMSO), and combinations thereof.
5. The composition as claimed in claim 4, wherein the composition comprises trehalose having a concentration in a range of 75-125 mM, and oxyresveratrol having a concentration in a range of 7.5 uM to 15 uM.
6. The composition as claimed in claim 5, wherein the composition comprises trehalose having a concentration of 100 mM, and oxyresveratrol having a concentration of 10 uM.
7. A process for preparing the composition as claimed in claim 1, said process comprising:

a) obtaining trehalose;
b) obtaining oxyresveratrol; and
c) contacting trehalose and oxyresveratrol, to obtain the composition.

8. A process for preparing the composition as claimed in claim 4, said process
comprising:
a) obtaining trehalose;
b) obtaining oxyresveratrol;
c) obtaining the at least one excipient; and
d) contacting trehalose, oxyresveratrol, and the at least one excipient, to obtain the composition.

9. The composition as claimed in any one of the claims 1-6, wherein the composition exhibits anti-ageing effects.
10. The composition as claimed in any one of the claims 1-6, wherein the composition exhibits skin-whitening effects.