FIELD OF INVENTION
[001] The present disclosure broadly relates to the field of cosmetics, in particular it discloses an anti-ageing composition. The present disclosure further discloses a process of preparing the anti-ageing composition.
BACKGROUND OF INVENTION
[002] Human skin is made up of several layers that is classified as epidermis,
dermis and subcutaneous tissue. The epidermis is the outermost layer of the skin
providing barrier to external environment. The dermis is the second layer of skin
containing various structural elements along with connective tissue. The presence of
three primary structural components in dermis such as collagen, elastin and GAGs
form an important support system called as extracellular matrix (ECM). The ECM
of the skin imparts strength and integrity to the skin.
[003] Collagen is one of the essential structural protein produced by fibroblasts
present in the dermis layer of the skin. Particularly, collagen is present in the ECM
that provides strength to skin and holding skin together. Collagen is produced and
recycled throughout the life.
[004] However, with increasing age, the rate of synthesis of collagen reduces. The
reduction in the rate of synthesis of collagen accelerates due to the increasing number
of enzymes such as matrix metalloproteinase (MMP). MMP cause the enhanced
degradation of collagen protein, subsequently leading to the appearance of fine lines
and wrinkles.
[005] Skin aging is a complex biological process that is influenced by combination
of endogenous or intrinsic (genetics, cellular metabolism, hormone and metabolic
processes) factors and exogenous or extrinsic (chronic light exposure, pollution,
ionizing radiation, chemicals, toxins) factors (Cevenini, H. et al. 2008 Sep
1;8(9):1393-405). The afore-mentioned factors leading to cumulative structural and
physiological changes in each layer of the skin which leads to the appearance of fine
lines and wrinkles.
[006] Various strategies have been deployed to reduce wrinkles and further
providing anti-ageing effect to the skin.

[007] US5371089A discloses a method and composition for ameliorating the adverse effects of aging, wherein the composition contains one or more 6-(substituted amino) purine cytokinins that can applied on mammalian skin to provide cosmetic appearance of mammalian skin without altering the growth rate and total proliferative capacity of mammalian skin cells.
[008] US20090196895A1 discloses a cosmetic preparation comprising a skin complex, wherein said skin complex comprising a liposome. The liposome comprising a mixture of cosmetic oil, extract of Plukenetia volubilis seeds, extract of Cynara scolymus leaves and hydrogenated retinol. The liposomes can be homogeneously dispersed in a gel network consisting of water and a gel-forming agent to provide said cosmetic preparation having an anti-ageing effect. [009] EP2939657A1 discloses an external dermal composition, wherein said external dermal composition comprising an aqueous medium as a base material and one or more members selected from the group consisting of L-ascorbic acid, an L-ascorbic acid derivative, and a salt thereof as an effective ingredient(s). The external dermal composition disclosed in the Application provides anti-ageing effect. [0010] However, several approaches that have been made to provide ant-ageing effect fails to provide desirable anti-ageing effect. Most of the products that are conventionally available in the market are unable to penetrate across the layers of skin to provide an enhanced anti-ageing effect.
[0011] Therefore, there remains a need in the art for an effective composition that exhibits enhanced anti-ageing properties and providing an accelerated increase in the synthesis of collagen.
SUMMARY OF THE INVENTION
[0012] In an aspect of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water;

(h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9. [0013] In an aspect of the present disclosure, there is provided a process for preparing the composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9, said process comprising: (i) obtaining aqua (and) glycerine (and) beta-Glucan; (ii) obtaining L-histidine; (iii) obtaining L-carnosine; (iv) obtaining 6-furfurylaminopurine; (v) obtaining L-citrulline; (vi) obtaining Cyprus rotundus root extract (and) glycerine (and) water; and vii) obtaining Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (viii) obtaining niacinamide; (ix) obtaining sodium hyaluronate; and (x) contacting aqua (and) glycerine (and) beta-Glucan, L-histidine, L-carnosine, 6-furfurylaminopurine, L-citrulline, Cyprus rotundus root extract (and) glycerine (and) water, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water, niacinamide, and sodium hyaluronate, to obtain the composition.
[0014] 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
[0015] 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.
[0016] Figure 1 depicts the effect of control, retinoic acid (RA), SAA-HCC and SAA-HCC2 in synthesising collagen, in accordance with an embodiment of the present disclosure.
[0017] Figure 2 depicts the comparison of gene expression fold change of collagen I in human dermal fibroblast (HDF) upon treating with control, SAA-HCC3, and retinoic acid (RA), in accordance with an embodiment of the present disclosure. [0018] Figure 3 depicts the comparison of collagen protein expressed by HDF upon treating with control, retinoic acid (RA) and SAA-HCC3, in accordance with an embodiment of the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0019] 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
[0020] 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.
[0021] L[X Teg\V_Xf pTq* pTaq TaW pg[Xq TeX hfXW gb eXYXe gb baX be gb `beX g[Ta baX
(i.e., to at least one) of the grammatical object of the article.
[0022] L[X gXe`f pVb`ce\fXq TaW pVb`ce\f\aZq TeX hfXW \a g[X \aV_hf\iX* bcXa fXafX*
meaning that additional elements may be included. It is not intended to be construed
Tf pVbaf\fgf bY ba_lq,

[0023] Throughout this specification, unless the context requires otherwise the word pVb`ce\fXq* TaW iTe\Tg\baf fhV[ Tf pVb`ce\fXfq TaW pVb`ce\f\aZq* j\__ UX 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. [0024] L[X gXe` p\aV_hW\aZq \f hfXW gb `XTa p\aV_hW\aZ Uhg abg _\`\gXW gbq, pCaV_hW\aZq TaW p\aV_hW\aZ Uhg abg _\`\gXW gbq TeX hfXW \agXechangeably. [0025] 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.
[0026] L[X gXe` pTg _XTfg baXq \f hfXW gb `XTa baX be `beX TaW g[hf \aV_hWXf individual components as well as mixtures/combinations.
[0027] @be g[X checbfX bY g[X ceXfXag W\fV_bfheX* g[X gXe` pTag\-TZX\aZq refers to a term that can be associated with skin-care products that helps to make the consumer look younger by reducing, masking or preventing signs of skin aging. [0028] The skin products as described herein can be in the form of ointments, serums, creams, lotions, or any other unit dosage forms such as sachets, and the like. [0029] As used herein, the term pX`b__\Xagq is a moisturizer that can applied directly to the skin to soothe and hydrate it9 p[h`XVgTagq \f T cosmetic ingredient that ceXiXagf _bff bY `b\fgheX* g[XeXUl eXgT\a\aZ g[X f^\a'f aTgheT_ `b\fgheX9 pcb_l`Xeq \f a large molecule made up of repeating sequences of identical smaller molecules; pf\_\VbaXfq eXYXef gb cb_l`Xef g[Tg \aV_hWX Tal flag[Xg\V Vb`cbhaW `TWX hc bY repeating units of siloxane, which is a chain of alternating silicon atoms and oxygen atoms, combined with carbon, hydrogen, and sometimes other elements. [0030] The terms pceXfXeiTg\iXfq* pV[X_Tgbefq* p`b\fghe\mTg\ba TZXagfq* TaW pfhafVeXXa TZXagfq eXYXe gb g[X ZXaXeT__l ^abja Vb`cbaXagf hfXW \a g[X Teg, [0031] 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.
[0032] 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. [0033] The plethora of anti-aging skin products that are available in the market contain ingredients that fail to provide adequate ant-ageing properties as they are unable to reach the dermal layer of skin. These ant-ageing skin products are further associated with various undesirable side effects such as acne and pimples. Also, usage of some anti-ageing products may lead to the formation of red patches and blisters on the skin followed by irritation. Further, said anti-ageing products may have a possibility of degrading the quality of skin by triggering series of allergic reactions.
[0034] Therefore, to overcome the afore-mentioned problems, the object of the present disclosure is to provide an anti-ageing composition. The object of the present disclosure further provides an anti-ageing composition that involves the amalgamation of various active ingredients exhibiting an increase in the synthesis of collagen in skin fibroblast cells. Subsequently, the composition of the present disclosure helps in maintaining the tensile strength of the skin and thereby delay the formation of wrinkles.
[0035] Also, the composition as disclosed in the present disclosure is safe to use and provides an adequate ant-ageing results without exhibiting any side effects. [0036] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9.

[0037] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9. In another embodiment of the present disclosure, L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.016 to 1:0.85. In yet another embodiment of the resent disclosure, L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.0165 to 1:0.80.
[0038] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein L-carnosine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.5. [0039] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein L-carnosine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.01 to 1:0.4. In another embodiment of the present disclosure, L-carnosine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.08 to 1:0.35. [0040] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine;

(c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.01 to 1:0.8 and wherein L-carnosine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.5. [0041] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein aqua (and) glycerine (and) beta-Glucan has a weight percentage in a range of 1.5-3.5% with respect to the composition, L-histidine has a weight percentage in a range of 0.02-0.05% with respect to the composition, L-carnosine has a weight percentage in a range of 0.05-0.2% with respect to the composition, 6-furfurylaminopurine has a weight percentage in a range of 0.0005-0.05% with respect to the composition, L-citrulline has a weight percentage in a range of 0.5-2% with respect to the composition, Cyprus rotundus root extract (and) glycerine (and) water has a weight percentage in a range of 0.2-1% with respect to the composition, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water has a weight percentage in a range of 0.05-0.2% with respect to the composition, niacinamide has a weight percentage in a range of 0.5-3% with respect to the composition and sodium hyaluronate has a weight percentage in a range of 0.005-0.02% with respect to the composition.
[0042] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water;

(h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein wherein aqua (and) glycerine (and) beta-Glucan has a weight percentage in a range of 1.8-2.8% with respect to the composition, L-histidine has a weight percentage in a range of 0.02-0.04% with respect to the composition, L-carnosine has a weight percentage in a range of 0.07-0.13% with respect to the composition, 6-furfurylaminopurine has a weight percentage in a range of 0.0006-0.04% with respect to the composition, L-citrulline has a weight percentage in a range of 0.8-1.7% with respect to the composition, Cyprus rotundus root extract (and) glycerine (and) water has a weight percentage in a range of 0.2-0.7% with respect to the composition, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water has a weight percentage in a range of 0.08-0.12% with respect to the composition, niacinamide has a weight percentage in a range of 0.8-2.5% with respect to the composition and sodium hyaluronate has a weight percentage in a range of 0.005-0.01% with respect to the composition.
[0043] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; (i) sodium hyaluronate, and (j) at least one excipient selected from a group consisting of emollient, humectants, polymers, silicones, preservatives, fragrance, vitamins, chelators, sunscreen agents, moisturization agents, and combinations thereof, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9. In another embodiment of the present disclosure, the humectant is glycerin, the silicones include dimethicone, the preservative is phenoxyethanol, the vitamin is tocopherol acetate, the chelator is disodium ethylenediaminetetraacetic (EDTA), the sunscreen agent is ethylhexyl methoxycinnamate, and the moisturization agent is glycerin.

[0044] In an embodiment of the present disclosure, there is provided a process for preparing the anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9, said process comprising: (i) obtaining aqua (and) glycerine (and) beta-Glucan; (ii) obtaining L-histidine; (iii) obtaining L-carnosine; (iv) obtaining 6-furfurylaminopurine; (v) obtaining L-citrulline; (vi) obtaining Cyprus rotundus root extract (and) glycerine (and) water; and (vii) obtaining Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (viii) obtaining niacinamide; (ix) obtaining sodium hyaluronate; and (x) contacting aqua (and) glycerine (and) beta-Glucan, L-histidine, L-carnosine, 6-furfurylaminopurine, L-citrulline, Cyprus rotundus root extract (and) glycerine (and) water, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water, niacinamide, and sodium hyaluronate, to obtain the composition.
[0045] In an embodiment of the present disclosure, there is provided a process for preparing the anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan having a weight percentage in a range 1.5-3.5% with respect to the composition; (b) L-histidine having a weight percentage in a range of 0.02-0.05% with respect to the composition; (c) L-carnosine having a weight percentage in a range of 0.05-0.2% with respect to the composition; (d) 6-furfurylaminopurine having a weight percentage in a range of 0.0005-0.05% with respect to the composition; (e) L-citrulline having a weight percentage in a range of 0.5-2% with respect to the composition; (f) Cyprus rotundus root extract (and) glycerine (and) water having a weight percentage in a range of 0.2-1% with respect to the composition; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water having a weight percentage in a range of 0.05-0.2% with respect to the composition; (h) niacinamide

having a weight percentage in a range of 0.5-3% with respect to the composition; and (i) sodium hyaluronate having a weight percentage in a range of 0.005-0.02% with respect to the composition, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9, said process comprising: (i) obtaining aqua (and) glycerine (and) beta-Glucan; (ii) obtaining L-histidine; (iii) obtaining L-carnosine; (iv) obtaining 6-furfurylaminopurine; (v) obtaining L-citrulline; (vi) obtaining Cyprus rotundus root extract (and) glycerine (and) water; and (vii) obtaining Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (viii) obtaining niacinamide; (ix) obtaining sodium hyaluronate; and (x) contacting aqua (and) glycerine (and) beta-Glucan, L-histidine, L-carnosine, 6-furfurylaminopurine, L-citrulline, Cyprus rotundus root extract (and) glycerine (and) water, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water, niacinamide, and sodium hyaluronate, to obtain the composition.
[0046] In an embodiment of the present disclosure, there is provided a process for preparing the anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; (i) sodium hyaluronate, and (j) at least one excipient selected from a group consisting of emollient, humectants, polymers, silicones, preservatives, fragrance, vitamins, chelators, sunscreen agents, moisturization agents, and combinations thereof, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9, said process comprising: (i) obtaining aqua (and) glycerine (and) beta-Glucan; (ii) obtaining L-histidine; (iii) obtaining L-carnosine; (iv) obtaining 6-furfurylaminopurine; (v) obtaining L-citrulline; (vi) obtaining Cyprus rotundus root extract (and) glycerine (and) water; and (vii) obtaining Malus Domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (viii) obtaining niacinamide; (ix) obtaining sodium hyaluronate; (x) obtaining at least one

excipient; and (xi) contacting aqua (and) glycerine (and) beta-Glucan, L-histidine, L-carnosine, 6-furfurylaminopurine, L-citrulline, Cyprus rotundus root extract (and) glycerine (and) water, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water, niacinamide, sodium hyaluronate, and the at least one excipient, to obtain the composition. [0047] In an embodiment of the present disclosure, there is provided a process for preparing the anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein the composition increases collagen synthesis. In another embodiment of the present disclosure, L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.01 to 1:0.8.
[0048] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein L-carnosine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.5 and wherein the composition increases collagen synthesis.
[0049] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9 and wherein

wherein aqua (and) glycerine (and) beta-Glucan has a weight percentage in a range of 1.5-3.5% with respect to the composition, L-histidine has a weight percentage in a range of 0.02-0.05% with respect to the composition, L-carnosine has a weight percentage in a range of 0.05-0.2% with respect to the composition, 6-furfurylaminopurine has a weight percentage in a range of 0.0005-0.05% with respect to the composition, L-citrulline has a weight percentage in a range of 0.5-2% with respect to the composition, Cyprus rotundus root extract (and) glycerine (and) water has a weight percentage in a range of 0.2-1% with respect to the composition, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water has a weight percentage in a range of 0.05-0.2% with respect to the composition, niacinamide has a weight percentage in a range of 0.5-3% with respect to the composition and sodium hyaluronate has a weight percentage in a range of 0.005-0.02% with respect to the composition, and wherein the composition increases collagen synthesis.
[0050] In an embodiment of the present disclosure, there is provided an anti-ageing composition comprising: (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate, and (j) at least one excipient selected from a group consisting of emollient, humectants, polymers, silicones, preservatives, fragrance, vitamins, chelators, sunscreen agents, moisturization agents, and combinations thereof, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.9, and wherein the composition increases collagen synthesis.
[0051] In an embodiment of the present disclosure, there is provided an anti-ageing composition as disclosed herein, wherein the composition may be applied onto the in the form of form of ointments, serums, creams, lotions, or any other unit dosage forms such as sachets, and the like.

[0052] Although the subject matter has been described in considerable detail with reference to certain examples and implementations thereof, other implementations are possible.
EXAMPLES
[0053] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply. The working and non-working examples as depicted in the forthcoming sections discloses the effect of the anti-ageing composition of the present disclosure on synthesis of collagen. Further, it highlights the criticality of the working percentages of different components in achieving the final anti-ageing composition of the present disclosure, wherein said anti-ageing composition comprising : (a) aqua (and) glycerine (and) beta-Glucan; (b) L-histidine; (c) L-carnosine; (d) 6-furfurylaminopurine; (e) L-citrulline; (f) Cyprus rotundus root extract (and) glycerine (and) water; and (g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; (h) niacinamide; and (i) sodium hyaluronate. It is further specified that the presence of each component within the disclosed weight ratio ranges and weight percentage ranges is critical so as to achieve anti-ageing composition exhibiting enhanced effect to increase the collagen synthesis in skin fibroblast cells.

[0055] For the purpose of the present disclosure, three samples were taken, sample 1= SAA-HCC; sample 2= SAA-HCC 2; sample 3= SAA-HCC3. Each afore-mentioned sample comprised set of active constituents such as a) aqua (and) glycerine (and) beta-Glucan; b) L-histidine; c) L-carnosine; d) 6-furfurylaminopurine; e) L-citrulline; f) Cyprus rotundus root extract (and) glycerine (and) water; and g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; h) niacinamide; and i) sodium hyaluronate, as disclosed in the present disclosure. The said active constituents of the present disclosure were combined to arrive at the final anti-ageing composition of the present disclosure.
[0056] All active constituents such as Cyprus rotundus root extract (and) glycerine (and) water [Actipone® Nutgrass (Motha) Root GW], Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water were commercially procured. Additionally, all other remaining constituents used in the present disclosure were also commercially procured. The afore-mentioned constituents used for arriving at the final composition of the present disclosure exhibits ant-ageing properties. Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water [I[lgb=X__LXVo FT_hf >b`Xfg\VTS Tf WXfVe\UXW [XeX\a in the disclosure, is a novel cosmetic active ingredient containing apple stem cell compounds encapsulated in liposomes prepared from soy phospholipids. The ingredient with its viscous appearance is used for skin rejuvenating properties. [0057] Table 1 as given below depicts the composition of I[lgb=X__LXVo FT_hf Domestica. Table 1: Composition of I[lgb=X__LXVo FT_hf >b`Xfg\VT

[0058] Sircol Collagen assay
[0059] Sircol collagen assay was carried out to monitor the ability of each sample of
the present disclosure (SAA-HCC, SAA-HCC 2 and SAA-HCC 3) to enhance the
synthesis of collagen in skin fibroblast cells.
[0060] Human dermal fibroblasts (HDF) were seeded in 12 well plates (from Costar)
in Dulbecco's Modified Eagle Medium (DMEM) high glucose containing 2% fetal
bovine serum (Gibco). The culture was incubated in 5% CO2 incubator and was
allowed to reach confluency, to form a fibroblast bed. HaCaT cells at a seeding
density of 0.11 million cells per well were seeded on the fibroblast bed. Keratinocyte
basal medium with supplements (Gibco) containing 0.03mM calcium chloride was
used while seeding the HaCaTs and the culture was maintained for 24 hrs for cell
attachment and acclimatization. Post 24 hrs, treatments were initiated. All treatments
were diluted in the keratinocyte basal medium with supplements containing 0.03mM
calcium chloride. After 48hrs of treatment, spent medium was harvested to determine
total collagen based on thX `TahYTVgheXerf cebgbVb_,
[0061] Gene Expression Study
[0062] To investigate the expression pattern of collagen, real time PCR (RT-PCR)
was performed. To this end, the cells were cultured in a similar fashion as described
in the sircol collagen assay. After 48 hrs of treatment cells were trypsinized and
pelleted down and then taken forward for RNA isolation. Total mRNA from cells
was extracted using a RNA mini kit (Ambion, Life Technologies). Following RNA
isolation, hexamer-primed reverse transcription was performed using the High
Capacity cDNA Reverse transcription kit (Applied Biosystems). Thereafter RT-PCR
against human collagen IA was performed using SYBR Green as reporter gene.
[0063] The primer used for collagen IA:
@bejTeW8 3r-CTGTTCTGTTCCTTGTGTAACTGTGTT
JXiXefX8 3r-GCCCCGGTGACACATCAA.
Method of preparing the composition of the present disclosure

[0064] The composition of the present disclosure was prepared by following steps:
[0065] Step 1: The constituents such as a) aqua (and) glycerine (and) beta-Glucan;
b) L-histidine; c) L-carnosine; d) 6-furfurylaminopurine; e) L-citrulline; f) Cyprus
rotundus root extract (and) glycerine (and) water; and g) Malus domestica fruit cell
culture extract (and) xanthan gum (and) glycerine (and) lecithin (and)
phenoxyethanol (and) water; h) niacinamide; and i) sodium hyaluronate having
disclosed weight percentage ranges as disclosed in the present disclosure, were
obtained
[0066] Step 2: The said constituents as disclosed in step 1 were contacted to each
other.
[0067] Step 3: The constituents were further mixed together in a blender at a room
temperature to provide the homogenous composition of the present disclosure.
Example 1
[0068] Working and non-working example
The table 2 as given below depicts the individual concentration of active constituents used for preparing the anti-ageing composition of the present disclosure. Each column depicts the concentration of active constituents in each sample used in the present disclosure, i.e. SAA-HCC, SAA-HCC-2 and SAA-HCC-3. It can be observed that the concentration of L-histidine and 6-furfurylaminopurine in SAA-HCC were 0.039% and 0.0007% respectively, in SAA-HCC2 were 0.28% and 0.0007% respectively and in SAA-HCC3 were 0.039% and 0.03% respectively. The concentration of L-histidine and 6-furfurylaminopurine in SAA-HCC and SAA-HCC3 were falling within the disclosed weight ratio range of the present disclosure, whereas in SAA-HCC 3, the concentration of L-histidine and 6-furfurylaminopurine were not falling in the disclosed wt. ratio range. Similarly, the concentration of L-carnosine in SAA-HCC and SAA-HCC3 were falling within the disclosed wt. ratio range of the present disclosure, whereas in SAA-HCC2 the concentration of L-carnosine was falling outside the disclosed weight ratio range of the present disclosure.

[0069] Therefore, it can be inferred from above observations that SAA-HCC and SAA-HCC 3 were considered as working examples of the present disclosure and SAA-HCC2 as a non-working example of the present disclosure. Table 2: Concentration of active constituents.
Sample 1: Sample 2: Sample 3:
Actives (%) SAA-HCC SAA-HCC 2 SAA-HCC 3
aqua (and) glycerine (and)
beta-glucan 2.5 2.5 2.5
L-histidine 0.039 0.28 0.039
L-carnosine 0.1 0.72 0.1
6-furfurylaminopurine 0.0007 0.0007 0.03
L-citrulline 1 1 1
Cyprus rotundus root extract (and) glycerine
(and) water 0.5 0.5 0.5
Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol
(and) water 0.1 0.1 0.1
Niacinamide 1 1 2
sodium hyaluronate 0.05 0.05 0.05
Water 94.7103 93.8493 93.681
Sample 1: Sample 2: Sample 3:
Actives (%) SAA-HCC SAA-HCC 2 SAA-HCC 3
aqua (and) glycerine (and)
beta-glucan 2.5 2.5 2.5
L-histidine 0.039 0.28 0.039
L-carnosine 0.1 0.72 0.1
6-furfurylaminopurine 0.0007 0.0007 0.03
L-citrulline 1 1 1
Cyprus rotundus root extract (and) glycerine
(and) water 0.5 0.5 0.5
Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol
(and) water 0.1 0.1 0.1
Niacinamide 1 1 2
sodium hyaluronate 0.05 0.05 0.05
Water 94.7103 93.8493 93.681
Example 2
[0070] Assessing collagen synthesis
[0071] Collagen is an essential protein present in extracellular matrix of the skin and
produced by the fibroblast present in the dermal layer of the skin. The presence of
collagen in ECM of the skin is very crucial as collagen helps in imparting integrity
and strength to the skin. With increasing age, the collagen protein gets degraded by
matrix metalloproteinase and collagen synthesis gets reduced. Subsequently, leading
to appearance of fine lines and wrinkles further leading to aging of the subject.
Therefore, the present disclosure discloses an anti-ageing composition leading to the
enhancement of the inherent ability of fibroblasts cells to synthesize collage in order

to retain the tensile strength of skin and subsequently leading to a delay in the
formation of wrinkles.
[0072] To detect the inherent ability to enhance the collagen synthesis in skin
fibroblast cells, each sample were tested by performing the cellular sircoll assay,
wherein human dermal fibroblasts (HDF) were treated with vehicle control (water),
retinoic acid (RA), SAA-HCC or SAA-HCC 2 in keratinocyte basal medium which
was further supplemented with 0.03mM CaCl2 for 48hrs and wherein SAA-HCC and
SAA-HCC 2 were diluted 1:100 in culture medium for the treatment.
[0073] Thereafter, the spent medium was collected, and the Sircoll Assay was
performed in order to detect the collagen secreted by the fibroblast cells.
[0074] It can be depicted from Figure 1 that SAA-HCC exhibited a 20% percentage
increase in amount of collagen synthesis as compared to SAA-HCC 2. Since the
increment of L-histidine and carnosine did not bring about any beneficial effect in
enhancing collagen synthesis, a third sample (sample 3 = SAA-HCC3) was tested in
the successive example.
Example 3
Collagen gene expression
[0075] In order to determine the collagen gene expression of the sample 3=SAA-HCC 3 of the present disclosure, the co-culture was either treated with vehicle control (water), retinoic acid (RA), and SAA-HCC 3. Cells were treated in keratinocyte basal medium which was supplemented with 0.03mM CaCl2 for 48hrs and thereafter it was trypsinized and pelleted down and further processed for RT-PCR.
[0076] Referring to Figure 2, it can be observed that SAA-HCC 3 exhibited a 0.5-fold increase in the gene expression of collagen as compared to control sample.
Example 4
Collagen protein expression
[0077] The expression of protein in SAA-HCC 3 was determined by either treating
the co-culture with vehicle control (water), retinoic acid (RA), and SAA-HCC 3.
Cells were treated in keratinocyte basal medium, supplemented with 0.03mM CaCl2
for 48h. Thereafter, the spent medium was collected, and the Sircoll Assay was

performed in order to detect the collagen secreted by the fibroblast cells. For this purpose, SAA-HCC 3 was diluted 1:100 in culture medium for the treatment. [0078] It can be observed from Figure 3, that SAA-HCC 3 exhibited an increase of 20% in expressing collagen protein as compared to control sample. [0079] Therefore, it can be inferred from Figure 1, 2 and 3 that composition utilized in SAA-HCC and SAA-HCC 3 shows an increase in the synthesis of collagen in skin fibroblast cells. Further, the composition of the present disclosure provides anti-ageing properties that helps in delaying in the formation of wrinkles and fine lines.
Advantages of the present disclosure
[0080] The present disclosure discloses an anti-ageing composition comprising aqua (and) glycerine (and) beta-Glucan; L-histidine; L-carnosine; 6-furfurylaminopurine; L-citrulline; Cyprus rotundus root extract (and) glycerine (and) water; and Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water; niacinamide; and sodium hyaluronate. The composition disclosed in the present disclosure can be deployed to increase the collagen synthesis by enhancing the inherent ability of fibroblasts cells present in the dermal layer of the skin. Further, said composition helps in retaining the tensile strength of the skin and delaying in the formation of wrinkles and fine line. The composition of the present disclosure can be utilized in providing cost-effective skin anti-ageing products.

I/We Claim:
1. An anti-ageing composition comprising:
a) aqua (and) glycerine (and) beta-Glucan;
b) L-histidine;
c) L-carnosine;
d) 6-furfurylaminopurine;
e) L-citrulline;
f) Cyprus rotundus root extract (and) glycerine (and) water; and
g) Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water;
h) niacinamide; and i) sodium hyaluronate,
wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a
range of 1:0.005 to 1:0.9.
2. The anti-ageing composition as claimed in claim 1, wherein L-histidine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.01 to 1:0.8
3. The anti-ageing composition as claimed in claim 1, wherein L-carnosine to 6-furfurylaminopurine has a weight ratio in a range of 1:0.005 to 1:0.5.
4. The anti-ageing composition as claimed in claim 1, wherein aqua (and) glycerine (and) beta-Glucan has a weight percentage in a range of 1.5-3.5% with respect to the composition, L-histidine has a weight percentage in a range of 0.02-0.05% with respect to the composition, L-carnosine has a weight percentage in a range of 0.05-0.2% with respect to the composition, 6-furfurylaminopurine has a weight percentage in a range of 0.0005-0.05% with respect to the composition, L-citrulline has a weight percentage in a range of 0.5-2% with respect to the composition, Cyprus rotundus root extract (and) glycerine (and) water has a weight percentage in a range of 0.2-1% with respect to the composition, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water has a weight percentage in a range of 0.05-0.2% with respect to the composition, niacinamide has a weight percentage in a range of 0.5-3% with

respect to the composition and sodium hyaluronate has a weight percentage in a range of 0.005-0.02% with respect to the composition.
5. The anti-ageing composition as claimed in claim 1, wherein the composition further comprises at least one excipient selected from a group consisting of emollient, humectants, polymers, silicones, preservatives, fragrance, vitamins, chelators, sunscreen agents, moisturization agents, and combinations thereof.
6. A process for preparing the composition as claimed in claim 1, said process comprising:

a) obtaining aqua (and) glycerine (and) beta-Glucan;
b) obtaining L-histidine;
c) obtaining L-carnosine;
d) obtaining 6-furfurylaminopurine;
e) obtaining L-citrulline;
f) obtaining Cyprus rotundus root extract (and) glycerine (and) water; and
g) obtaining Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water;
h) obtaining niacinamide;
i) obtaining sodium hyaluronate; and
j) contacting aqua (and) glycerine (and) beta-Glucan, L-histidine, L-carnosine, 6-furfurylaminopurine, L-citrulline, Cyprus rotundus root extract (and) glycerine (and) water, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water, niacinamide, and sodium hyaluronate, to obtain the composition.
7. The process for preparing the composition as claimed in claim 5, said process
comprising:
a) obtaining aqua (and) glycerine (and) beta-Glucan;
b) obtaining L-histidine;

c) obtaining L-carnosine;
d) obtaining 6-furfurylaminopurine;
e) obtaining L-citrulline;
f) obtaining Cyprus rotundus root extract (and) glycerine (and) water; and
g) obtaining Malus Domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water;
h) obtaining niacinamide;
i) obtaining sodium hyaluronate;
j) obtaining at least one excipient; and
k) contacting aqua (and) glycerine (and) beta-Glucan, L-histidine, L-carnosine, 6-furfurylaminopurine, L-citrulline, Cyprus rotundus root extract (and) glycerine (and) water, Malus domestica fruit cell culture extract (and) xanthan gum (and) glycerine (and) lecithin (and) phenoxyethanol (and) water, niacinamide, sodium hyaluronate, and 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 increases collagen synthesis.