FIELD OF INVENTION
[001] The present disclosure relates to the field of cosmetic formulations in general and skin aging formulations in particular. There is provided a composition comprising red algal extract and 6, 7, 4'-trihydroxy isoflavone.
BACKGROUND OF INVENTION
[002] Skin aging is a natural process that is observable visually in form of fine lines, wrinkles and pigmentation on the skin. The skin is supported by the extracellular matrix (ECM). The constituents of the ECM of the skin include connective tissue proteins collagen and elastin, among others. These provide elasticity to the skin. [003] The process of skin aging is affected by a combination of both internal and external factors. External causes of skin aging include exposure to UV radiation, pollution, chronic light exposure, toxins and chemicals among others. The internal processes that affect skin aging are multifactorial including, the effect of hormones and metabolic changes within the body.
[004] Among the intrinsic causes of skin aging is the activity of proteases that specifically degrade proteins present in the skin such as collagen. These proteases have shown to increase with increasing age and thus, this process of degradation of the ECM proteins leads to the loss of skin elasticity (Roberta et. al, 1999, Free Radical Biology and Medicine, 27 (7-8), 729-737). Matrix metalloproteinases are among the major proteases falling within this class of proteases and they degrade collagen I.
[005] US6955816 B2 discloses anti-aging skin care compositions comprising Vitamin Bl, Vitamin B5, Vitamin C, N-acetyl-cysteine and, optionally, lipoic acid for therapeutically or prophylactically treating the consequences of aging on the condition or appearance of the skin.

[006] FR2838340 A1 discloses the use of an extract of brown seaweed Pelvetia canaliculata as active ingredient in cosmetic and dermatological compositions. [007] PCT/US2008/006207 discloses a composition comprising a combination of: Polygonum fagopyrum seed extract, Chlorella vulgaris extract, palmitoyl wheat protein hydrolysate, algae extract, and tripeptide for aiding in improving the firmness of the skin, lifting the skin, decreasing skin sagging, and decreasing visible signs of aging including one or more of wrinkles, fine lines, uneven skin tone and age spots. [008] Although compositions to combat skin aging and to reduce the signs of skin aging are available, there are only a few compositions that aim to target specific contributors of skin aging. Thus, there is a need to address the multifaceted problem of skin aging in an effective and targeted manner.
SUMMARY OF INVENTION
[009] In an aspect of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, wherein the red algal extract weight percentage in said composition is in the range of 0.000002%-0.000009%, 6, 7, 4’- trihydroxy isoflavone weight percentage in said composition is in the range of 0.000006%-0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0010] In an aspect of the present disclosure, there is provided a process for preparing a composition comprising: (i) red algal extract; and (ii) 6, 7, 4’- trihydroxy isoflavone, wherein the red algal extract weight percentage in said composition is in the range of 0.000002%-0.000009%, 6, 7, 4’- trihydroxy isoflavone weight percentage in said composition is in the range of 0.000006%-0.00003%, said method comprising: (a) obtaining the red algal extract; (b) obtaining the 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said

composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio
in said composition is in the range of 1:2-1:3.
[0011] In an aspect of the present disclosure, there is provided a method to inhibit
skin aging, said method comprising: (a) obtaining a composition comprising: (i) red algal extract; and (ii) 6, 7, 4’-trihydroxy isoflavone, wherein the red algal extract weight percentage in said composition is in the range of 0.000002%-0.000009%, 6, 7, 4’- trihydroxy isoflavone weight percentage in said composition is in the range of 0.000006%-0.00003%, and the red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0012] 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
[0013] 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.
[0014] Figure 1 shows the effect of red algal extract within a concentration range on the viability of human dermal fibroblast cells, in accordance with an embodiment of the present disclosure.

[0015] Figure 2 shows the effect of 6, 7, 4’- trihydroxy isoflavone (THIF) within a concentration range on the viability of human dermal fibroblast cells, in accordance with an embodiment of the present disclosure.
[0016] Figure 3a shows the HPLC chromatogram of 4-hydroxy benzoic acid standard, in accordance with an embodiment of the present disclosure.
[0017] Figure 3b shows the HPLC chromatogram of red algal chloroform: methanol extract, in accordance with an embodiment of the present disclosure.
[0018] Figure 4a shows the mass chromatogram of 4-hydroxy benzoic acid standard, in accordance with an embodiment of the present disclosure.
[0019] Figure 4b shows the mass chromatogram of red algal chloroform: methanol extract, in accordance with an embodiment of the present disclosure.
[0020] Figure 5 shows the effect of the combination of red algal extract and 6, 7, 4’-trihydroxy isoflavone (THIF), in accordance with an embodiment of the present disclosure.
DETAILED DESCRIPTION OF THE INVENTION
[0021] 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
[0022] 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.
[0023] 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.
[0024] 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”.
[0025] 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.
[0026] The term “including” is used to mean “including but not limited to”.
“Including” and “including but not limited to” are used interchangeably.
[0027] Carriers are substances that serve as mechanisms to improve the delivery and
the effectiveness of drugs.
[0028] A diluent (also referred to as filler, dilutant, or thinner) is a diluting agent.
[0029] 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,
preservatives, emollients, and combinations thereof.
[0030] Unless defined otherwise, all technical and scientific terms used herein have
the same meaning as commonly understood by one of ordinary skill in the art to
which this disclosure belongs. Although any methods and materials similar or
equivalent to those described herein can be used in the practice or testing of the
disclosure, the preferred methods, and materials are now described. All publications
mentioned herein are incorporated herein by reference.
[0031] Skin aging is affected by multiple factors. The extracellular matrix (ECM)
plays a pivotal role in the process of skin aging. A major contributing factor to skin
aging is the activity of proteases which degrades the ECM. Matrix Metalloproteases 1

(MMP 1) is among one of the major proteases which plays a predominant role in degrading the proteins present in the ECM, specifically collagen I. It has been observed that the levels of MMP1 increase with age. Additionally, irritation, inflammation and environmental factors also elevate MMP levels. A decrease in MMP1 level would decrease the collagen degradation promoting a youthful appearance. The present disclosure provides a solution to the problem of skin aging by inhibiting the levels of MMP1 proteases, thus delaying the process of aging. [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] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%, weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0034] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; and (b) weight percentage of 6, 7, 4’-trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0035] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a

weight percentage of 0.0000075% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2-1:3. [0036] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002%, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2-1:3.
[0037] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; and (b) 6, 7, 4’-trihydroxy isoflavone having a weight percentage in the range of 0.000006% -0.00003% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0038] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0039] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0040] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red

algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, and said red algal extract is obtained from the species Kappaphycus alvarezii.
[0041] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said red algal extract is obtained from the species Kappaphycus alvarezii, and concentration of 4-hydroxybenzoic acid in said algal extract is in the range of 0.02- 0.06%.
[0042] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said red algal extract is obtained from the species Kappaphycus alvarezii, and concentration of 4-hydroxybenzoic acid in said algal extract is 0.04 %. [0043] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% -0.000009%; and (b) weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.

[0044] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5. [0045] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0046] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of is 0.0000075% in said composition; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is 1:2-1:3.
[0047] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3. [0048] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage

in the range of 0.000002% - 0.000009% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0049] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0050] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0051] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging for inhibiting skin aging comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, red wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’-trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, and said red algal extract is obtained from the species Kappaphycus alvarezii.
[0052] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging for inhibiting skin aging comprising: (a) red algal extract; (b)

6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, red wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’-trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said red algal extract is obtained from the species Kappaphycus alvarezii, and concentration of 4-hydroxybenzoic acid in said algal extract is in the range of 0.02- 0.06%.
[0053] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging for inhibiting skin aging comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent, and (e) at least one excipient, red wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’-trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said red algal extract is obtained from the species Kappaphycus alvarezii, and concentration of 4-hydroxybenzoic acid in said algal extract is 0.04 %.
[0054] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract and; (ii) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’-trihydroxy isoflavone to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%, weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.

[0055] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract and; (ii) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’-trihydroxy isoflavone to obtain said composition, wherein weight percentage of the red algal extract is in said composition in the range of 0.000002% -0.000009%,weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0056] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition and; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0057] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (i) red algal extract having a weight percentage of 0.000003% in said composition and; (ii) 6, 7, 4’-trihydroxy isoflavone having a weight percentage of 0.0000075%, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0058] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage

of 0.000008% in said composition and; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2-1:3.
[0059] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition and; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0060] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage of 0.000003% in said composition and; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0061] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage of 0.000008% in said composition and; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; and (c)

contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0062] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract and; (ii) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7 ,4’- trihydroxy isoflavone; and (c) contacting the red algal extract and 6, 7, 4’-trihydroxy isoflavone to obtain said composition, red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, wherein said red algal extract is obtained from the species Kappaphycus alvarezii.
[0063] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (i) red algal extract; (ii) 6, 7, 4’- trihydroxy isoflavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining the at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3. [0064] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (i) red algal extract; (ii) 6, 7, 4’- trihydroxy isoflavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f)

contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[0065] In an embodiment of the present disclosure, there is provided a method of preparing a composition for inhibiting skin aging comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0066] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage of 0.000003% in said composition; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said

composition, and the red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0067] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage of 0.000008% in said composition; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition 1:2-1:3.
[0068] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0069] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage in the range of 0.000003% in said composition; (ii) 6, 7, 4’- trihydroxy isoflavone

having a weight percentage in the range of 0.0000075% in said composition; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’-trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5. [0070] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract having a weight percentage of 0.000008% in said composition; (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (e) obtaining the at least one excipient; and (f) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0071] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (i) red algal extract; (ii) 6, 7, 4’- trihydroxy isoflavone; (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, said method comprising: (a) obtaining the red algal extract; (b) obtaining 6, 7, 4’- trihydroxy isoflavone; (c) obtaining the at least one carrier; (d) obtaining at least one diluent; (c) obtaining the at least one excipient; and (d) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one carrier, the at least one diluent, and the at least one excipient, to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%;

weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, and said red algal extract is obtained from Kappaphycus alvarezii.
[0072] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract, and (ii) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0073] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract, and (ii) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006%-0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0074] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition, and (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.

[0075] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage of 0.000003% in said composition, and (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0076] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage of 0.000008% in said composition, and (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0077] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising; (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition, and (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0078] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising; (i) red algal extract having a weight percentage of 0.000003% in said composition, and (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio

in said composition is in the range of 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0079] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising; (i) red algal extract having a weight percentage of 0.000008% in said composition, and (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0080] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract, and (ii) 6, 7, 4’- trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006%-0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said red algal extract is obtained from Kappaphycus alvarezii.
[0081] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition, (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.

[0082] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage of 0.000003% in said composition, (ii) 6, 7, 4’-trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0083] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage of 0.000008% in said composition, (ii) 6, 7, 4’-trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0084] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition, (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0085] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage of 0.000003% in said composition, (ii) 6, 7, 4’-

trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0086] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage of 0.000008% in said composition, (ii) 6, 7, 4’-trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5; and (b) contacting said composition with skin, wherein said composition inhibits skin aging.
[0087] In an embodiment of the present disclosure, there is provided a method to inhibit skin aging comprising: (a) obtaining a composition comprising: (i) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition, (ii) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% - 0.00003% in said composition, (iii) at least one carrier; (iv) at least one diluent; and (v) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3; and (b) contacting said composition with skin, wherein said red algal extract is obtained from Kappaphycus alvarezii.
[0088] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract, and (b) 6,7,4’-trihydroxy isoflavone, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002%-0.000009%; weight percentage of 6, 7, 4’-trihydroxy isoflavone in said composition is in the range of 0.000006%-0.00003%, and the red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition

is in the range of 1:2-1:3, and said red algal extract is obtained from Kappaphycus alvarezii.
[0089] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract, and (b) 6, 7, 4’-trihydroxy isoflavone, weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5. [0090] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition, and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% -0.00003% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0091] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition, and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0092] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition, and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0093] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight

percentage in the range of 0.000002% - 0.000009% in said composition, and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006% -0.00003% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0094] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition, and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0095] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[0096] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract; (b) 6,7,4’-trihydroxy isoflavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%, weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, and said red algal extract is obtained from Kappaphycus alvarezii. [0097] In an embodiment of the present disclosure, there is provided a composition for inhibiting skin aging for inhibiting skin aging comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the red algal extract in said

composition is in the range of 0.000002% - 0.000009%, weight percentage of 6, 7, 4’-trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said red algal extract is obtained from the species Kappaphycus alvarezii, and concentration of 4-hydroxybenzoic acid in said algal extract is in the range of 0.02- 0.06%.
[0098] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; (b) 6, 7, 4’-trihydroxy isoflavone having a weight percentage in the range of 0.000006% -0.00003% in said composition; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[0099] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage in the range of 0.000003% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.0000075% in said composition; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[00100] In an embodiment of the present disclosure, there is provided a
composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.

[00101] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; (b) 6, 7, 4’-trihydroxy isoflavone having a weight percentage in the range of 0.000006% -0.00003% in said composition; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[00102] In an embodiment of the present disclosure, there is provided a composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[00103] In an embodiment of the present disclosure, there is provided a
composition for use in preparing formulations comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (c) at least one carrier; (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5. [00104] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of

0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[00105] In an embodiment of the present disclosure, there is provided a method
of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’-trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% -0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[00106] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[00107] In an embodiment of the present disclosure, there is provided a method
of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’-trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% -0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition

is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
[00108] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’-trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[00109] In an embodiment of the present disclosure, there is provided a method
of preparing a composition comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
[00110] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage in the range of 0.000002% - 0.000009% in said composition; and (b) 6, 7, 4’-trihydroxy isoflavone having a weight percentage in the range of 0.000006% -0.00003% in said composition, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.

[00111] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’-trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[00112] In an embodiment of the present disclosure, there is provided a method
of preparing a composition comprising: (a) red algal extract having a weight percentage of 0.000008% in said composition; and (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5.
[00113] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution A is prepared in an organic solvent.
[00114] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution A is prepared in an inorganic solvent.

[00115] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution B is prepared in an organic solvent.
[00116] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; and (b) 6, 7, 4’- trihydroxy isoflavone, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; and (c) contacting solution A and solution B to obtain said composition, wherein said solution B is prepared in an inorganic solvent.
[00117] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2- 1:3, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6, 7, 4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition.
[00118] In an embodiment of the present disclosure, there is provided a method
of preparing a composition comprising: (a) red algal extract; (b) 6, 7, 4’- trihydroxy isoflavone; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein weight percentage of the red algal extract in said composition is in the range of 0.000002% - 0.000009%; weight percentage of 6, 7, 4’- trihydroxy

isoflavone in said composition is in the range of 0.000006% - 0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is 1:2.5, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition.
[00119] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage in the range of 0.000002%-0.000009% in said composition; (b) 6, 7, 4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006%-0.00003% in said composition; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition. [00120] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage of 0.000003% in said composition; (b) 6,7,4’- trihydroxy isoflavone having a weight percentage of 0.0000075% in said composition; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein red algal extract to 6,7,4’-trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition.
[00121] In an embodiment of the present disclosure, there is provided a method
of preparing a composition comprising: (a) red algal extract having a weight

percentage of 0.000008% in said composition; (b) 6,7,4’- trihydroxy isoflavone having a weight percentage of 0.00002% in said composition; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition.
[00122] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage in the range of 0.000002%-0.000009% in said composition; (b) 6,7,4’- trihydroxy isoflavone having a weight percentage in the range of 0.000006%-0.00003% in said composition; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition.
[00123] In an embodiment of the present disclosure, there is provided a method of preparing a composition comprising: (a) red algal extract having a weight percentage in the range of 0.000003% in said composition; (b) 6,7,4’- trihydroxy isoflavone having a weight percentage in the range of 0.0000075% in said composition; (c) at least one carrier, (d) at least one diluent; and (e) at least one excipient, wherein red algal extract to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5, said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers, diluents, and excipients; and (d) contacting solution A, solution B, and solution C to obtain said composition.

[00124] In an embodiment of the present disclosure, there is provided a method of
preparing a composition comprising: (a) red algal extract having a weight percentage
in the range of 0.000008% in said composition; (b) 6,7,4’- trihydroxy isoflavone
having a weight percentage in the range of 0.00002% in said composition; (c) at least
one carrier, (d) at least one diluent; and (e) at least one excipient, wherein algal extract
to 6,7,4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2.5,
said method comprising: (a) obtaining a solution A of red algal extract; (b) obtaining a
solution B of 6,7,4’- trihydroxy isoflavone; (c) obtaining a solution C of carriers,
diluents, and excipients; and (d) contacting solution A, solution B, and solution C to
obtain said composition.
[00125] In an embodiment of the present disclosure, there is provided a composition
described herein, wherein said composition inhibits skin aging.
[00126] In an embodiment of the present disclosure, there is provided a composition
described herein, wherein said composition inhibits skin aging by inhibiting collagen
degrading enzymes.
[00127] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be for topical application.
[00128] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in the form of an aerosol.
[00129] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in the form of a lotion.
[00130] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in form of an oil.
[00131] In an embodiment of the present disclosure, there is provided a composition
as described herein, wherein said composition can be in the form of a serum.
[00132] Although the subject matter has been described with reference to specific
embodiments, this description is not meant to be construed in a limiting sense.
Various modifications of the disclosed embodiments, as well as alternate

embodiments of the subject matter, will become apparent to persons skilled in the art upon reference to the description of the subject matter. It is therefore contemplated that such modifications can be made without departing from the spirit or scope of the present subject matter as defined.
EXAMPLES
[00133] 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 vary.
Example 1
[00134] Material and Methods: Adult human dermal fibroblasts (HDFa) were cultured in Medium 106 (Gibco)_containing growth factors (LSGS Supplement). The media was replaced every alternate day till >80% confluence was attained. The cells were dislodged using with 0.25% trypsin EDTA solution and sub-cultured by seeding 3500 cells/cm2 of the flask surface. The cultures were tested for the absence of Mycoplasma by MycoAlert Mycoplasma Detection Kit (Lonza) once every month.
[00135] For cytotoxicity assay, the cells were seeded at a density of 8750 viable cells per well (100µL/well) in M106 medium containing growth factors in 96 well cell culture plates. After approximately 48 hour incubation, when the HDFa cells appeared to form a confluent monolayer in the wells, the media was removed and 13750 HaCaT

cells were layered onto the HDFa matrix (100 µL/well) in M154 CF-PRF (Gibco) media containing 30 µM CaCl2. The plates were further incubated overnight prior to treatment with the test ingredients at the desired concentrations, along with appropriate vehicle and sterility controls (wells with media only, without cells). The plates were incubated in a humidified incubator at 37°C and 5% CO2 for 48 hours, after which the media from the plates was removed and replaced with 200µL of media, containing 10% (20 µL) of Cell Titre Aqueous Solution (MTS Reagent). The plates were incubated at 37°C for a further 5-6 hour followed by reading at 490 nm using a Spectramax Reader.
[00136] MMP-1 protein estimation assay: (i) HDFa-HaCaT co-culture- HDFa cells were seeded at 70,000 cells/well in 12-well cell culture plates and incubated for 48 hrs in M106 containing LSGS Growth Factors. Upon HaCaT addition (0.11 million/well), the media was changed to M154 CF-PRF containing 30 µM CaCl2 and the plates were incubated for 24 hours before the test compounds (in 1 ml media per well) were added at the concentrations mentioned. The compound dilutions were prepared in M154 CF-PRF containing 30 µM CaCl2. The treated and control cells were incubated for 48 hours at 37°C in a humidified incubator in a 5% CO2 environment.
[00137] Prior to conducting the MMP-1 assay, the culture supernatants were collected 48 hours post- incubation. The plates were centrifuged briefly (approximately 5 min) at 300 g to remove any cellular debris. 50 µL of the clarified culture supernatant was collected in 96-well U-bottom polypropylene plates. The tubes and plates were stored at -80°C till they were processed further in assays.
[00138] Plates were thawed on ice on the day MMP-1 estimation was to be performed. The assay was carried out using the Human MMP-1 ELISA Kit, Sigma, Cat# RAB0361. Before starting the ELISA assay, all the kit components were allowed to reach room temperature. Samples were diluted 40-fold in 96-wells deep well block by transferring 5µl of sample to 195µL of 1X assay diluent buffer supplied with the kit. The contents were mixed thoroughly with a multichannel pipette to ensure uniformity

of the suspension. Lyophilized Human MMP-1 Protein Standard vial supplied with the kit was briefly centrifuged and 400 μL of 1X Assay Diluent was added and mixed to prepare a 0.1 μg/mL standard. 5 µl of 0.1 µg/mL MMP-1 standard was added to 995 µL of 1X assay diluent to achieve 500 pg/mL concentration. 2-fold serial dilution from 500 pg/ml stock was carried out in assay diluent buffer upto 7.8 pg/ml for the standard curve. 100µl of samples (n=3) and MMP-1 standard dilutions (n=4) along with 100µl of only assay diluent (which serves as blank) was transferred to Human MMP-1 Antibody-coated ELISA Plate and incubated for 2.5 hours at room temperature with gentle shaking covering the plate with ELISA plate sealer. [00139] Meanwhile, the biotinylated Detection Antibody vial was briefly centrifuged and 100 μL of 1X Assay Diluent Buffer was added to prepare a detection antibody concentrate (80X). The detection antibody concentrate was then diluted by 80-fold with 1X assay diluent buffer for the final use.
[00140] 20X Wash Buffer was diluted to obtain adequate volume of 1X wash buffer. The solutions from the ELISA plates were then decanted out and washed 4 times with 1X wash buffer by filling each well with 300 µl of wash buffer using a multichannel pipette. Complete removal of liquid at each step was made sure by blot-drying the plates. 100 µl of 1X prepared biotinylated Detection Antibody was added to each well and the plates were incubated for 1 hour at room temperature with gentle shaking. After incubation, the detection antibody solution was discarded from the plate by decanting and the wash step was repeated with 1X wash buffer and the plates were dried by tapping against paper-towel. The HRP-Streptavidin concentrate vial was briefly spun and pipetted up and down to mix gently before use, as precipitates may form during storage. HRP-Streptavidin concentrate was then diluted 440-fold with 1X Assay Diluent Buffer before use.
[00141] 100µl of prepared 1X HRP-Streptavidin solution was then added to each well and incubated for 45 minutes at room temperature with gentle shaking. After incubation, the HRP-Streptavidin solution was discarded from the plate by decanting

and the wash step was repeated with 1x wash buffer and the plates were blot-dried. 100µl of ELISA Colorimetric TMB Reagent was added to each well and plates were incubated for 30 minutes at room temperature in the dark with gentle shaking. 50µl of Stop Solution was added to each well and the absorbance was measured at 450 nm immediately using FlexStation 3 (Molecular Devices).
Example 2
Identification and estimation of biomarker in Kappaphycus alvarezii extract [00142] For the purposes of fingerprint analysis for species identification, 4-hydroxybenzoic acid was identified as the appropriate biomarker (Figure 3a) and quantified in the Kappaphycus alvarezii extract (Figure 3b). Preparation of Kappaphycus alvarezii extract: 25.0 g of Kappaphycus alvarezii was taken and the total plant was ground to a coarse powder. This was loaded in the wide thimble tube. 250 ml (167mL Chloroform + 83mL of Methanol) of chloroform: Methanol solvent was taken in a 500 ml RB flask. The soxhlet apparatus was set up and kept for heating in a water bath at a temperature of 60-70°C for a period of 5-6 hours. After the extraction, the extract was filtered using a Whatmann filter paper No. 1 and collected in a pre-weighed 250 ml round bottomed flask. The chloroform: methanol extract was continuously concentrated in the rotary-evaporator at reduced pressure of 45-50 mbar and at a temperature of 40-50°C. The extract was thoroughly dried and allowed to cool. It was labelled and stored in a refrigerator at 4°C.
[00143] The 4-hydroxybenzoic acid standard was prepared by weighing 10.21 mg of standard was weighed accurately in a 10.0 mL volumetric flask, 6.0 mL of diluent was added and dissolved by sonication and made up to the volume with the diluent. 1.0 mL of above solution was further diluted to 10.0 mL with diluent.
[00144] The Kappaphycus alvarezii sample was prepared for HPLC analysis by weighing 50.13mg of sample in a 5.0mL volumetric flask, 3mL of methanol was the

added to this and dissolved by sonication. It was diluted up to the volume with
methanol.
[00145] HPLC analysis was carried out using the following parameters:
Preparation of Mobile phase: Mobile phase-A: 1% acetic acid in water, Mobile phase
–B: Methanol
Chromatographic conditions:
Column : Inertsil ODS C-18, 250*4.6mm, 5µm
Flow : 1.0mL/min
Pump mode : Gradient
Injection Volume: 50µL
Detector : DAD
Wavelength : 254nm
Column Temperature: Ambient
Run time : 50 mins
Diluent: Water: Methanol (20:80)
Solvent ratios are shown in Table 1.

Table 1
Time A B
(mins) (%) (%)
0.0 90 10
6.0 90 10
25.0 30 70
5
40.0 30 70
45.0 90 10
50.0 90 10
[00146] Based on the experimental findings
obtained from both HPLC and mass spectrometry analyses, it was found that the percentage of 4-hydroxybenzoic acid in Kappaphycus alvarezii chloroform: methanol extract was 0.04%. (Figures 3a, 3b, 4a,4b).
Example 3
Cytotoxicity analysis of actives
[00147] To determine the concentration ranges appropriate for further analyses of actives on cellular functions, cytotoxicity experiments were conducted for both the algal extract and THIF.
[00148] A range of 0.00000009%- 0.00002% of Kappaphycus alvarezii extract was tested on the HDFa-HaCaT co-culture described above (Figure 1). The assay showed that overall, the algal extract was non-toxic at most of the concentrations tested with an average viability at 103%. A minor decrease in cell viability was observed at 0.00002% at 98%. Based on these results, a non-toxic range of 0.000001%-0.000008% was chosen for further analyses.
[00149] A range of 0.021%- 2.7% of THIF was tested for cytotoxic effects. As seen in Figure 2, cell viability for most of the concentrations tested was at approximately 100%, at the highest concentration used i.e. 2.7%, the cell viability dropped partially

to 98%. From the observations obtained from these data, a non-toxic concentration range of 0.0000025%-0.00002% was chosen for further analyses.

Example 4
The combination of Kappaphycus alvarezii and THIF demonstrates synergism in
inhibiting skin aging
[00150] As discussed previously, MMP1 proteases contribute to skin aging by degrading extracellular matrix proteins such as collagen. Therefore, in order to quantify the effect of the actives under study in promoting skin again, the inhibition of MMP1 protein was quantified using the protocol described above.
[00151] As seen in Figure 5 the individual actives of Kappaphycus alvarezii extract and THIF did not significantly inhibit MMP1 protein production. A 10-fold difference of the concentrations of Kappaphycus alvarezii extract (in the range of 0.0000007%-0.000007%) in isolation did not demonstrate any fold change in levels across the concentration range. Overall, Kappaphycus alvarezii extract in isolation was able to supress the levels of MMP1 very slightly as compared to the control exhibiting a 0.8 fold decrease in protein levels.
[00152] Similarly, THIF was also supplied in isolation at the concentrations of 0.000135%, 0.00027% and 0.00054%. As evident from Figure 5, THIF, like the algal extract, did not demonstrate any significant effect in changing the protein level of MMP1. The levels of the protein were close to control levels, and on the contrary, showed a 1.2 fold increase in protein levels.
[00153] As Kappaphycus alvarezii extract was able to marginally repress MMP1 protein levels, the combination of Kappaphycus alvarezii extract with THIF was tested to determine whether THIF could enhance the inhibitory effect of the algal extract. Three concentrations were attempted for the combination of the two actives at 0.000008% Kappaphycus alvarezii extract (K) and 0.00002% THIF (T), 0.000003% K and 0.0000075% T, and 0.000001% K and 0.0000025% T.
[00154] Remarkably, the combination of K and T at the concentrations of 0.000008% K and 0.00002% T, and 0.000003% K and 0.0000075% T, showed significantly enhanced ability to inhibit the levels of MMP1 protein. This is an unexpected result as

individually, neither Kappaphycus alvarezii extract nor THIF showed any significant ability to inhibit MMP1 protein levels. The combination of the two, however, work synergistically to affect protein levels thereby delaying the process of skin aging through the degradation of the extracellular matrix.
[00155] Further, this combinatorial inhibitory effect of the actives is only apparent at
very specific concentrations within a 1:2-1:3 w/w concentration range. At a
concentration of 0.000001% K and 0.0000025% T, the synergistic inhibitory effect is not observed. Thus, a mere admixture of the two components is not sufficient to yield the observed synergistic effect.
[00156] Overall, the present disclosure provides a composition of red algal extract, and 6, 7, 4’- trihydroxy isoflavone (THIF), which exhibits a surprising and unexpected synergistic effect on the inhibition of MMP1 protein levels. This apparent synergism of the said combination is not readily apparent at all concentration combinations (as discussed previously), suggesting that the results are not a result of mere admixture of the three or routine in nature. Further, the identification of this novel combination to manage skin aging cannot be arrived at without undue experimentation, and is not obvious to a person skilled in the art.

I/We Claim:
1. A composition for inhibiting skin aging comprising:
(a) red algal extract; and
(b) 6, 7, 4’- trihydroxy isoflavone,
wherein the red algal extract weight percentage in said composition is in the range of 0.000002%-0.000009%, 6, 7, 4’- trihydroxy isoflavone weight percentage in said composition is in the range of 0.000006%-0.00003%, and the red algal extract to 6, 7, 4’- trihydroxy isoflavone w/w ratio in said composition is in the range of 1:2-1:3.
2. The composition as claimed in claim 1, wherein the red algal extract to 6, 7, 4’-trihydroxy isoflavone w/w ratio in said composition is 1:2.5.
3. The composition as claimed in any of the claims 1-2, wherein,

(a) the red algal extract weight percentage is 0.000003%; and
(b) 6, 7, 4’- trihydroxy isoflavone weight percentage is 0.0000075%.
4. The composition as claimed in any of the claims 1-2, wherein,
(a) the red algal extract weight percentage is 0.000008%; and
(b) 6, 7, 4’- trihydroxy isoflavone weight percentage is 0.00002%.

5. The composition as claimed in any of the claims 1-4, further comprising: (a) at least one additive; (b) at least one diluent, and (c) at least one excipient.
6. The composition as claimed in claim 1, wherein said red algal extract is obtained from Kappaphycus alvarezii.
7. A process for preparing a composition, as claimed in any of the claims 1-4, comprising:
(a) obtaining the red algal extract;

(b) obtaining 6, 7, 4’- trihydroxy isoflavone; and
(c) contacting the red algal extract and 6, 7, 4’- trihydroxy isoflavone to obtain said composition.
8. A process for preparing a composition, as claimed in any of the claims 1-5,
comprising the steps of:
(a) obtaining the red algal extract;
(b) obtaining 6,7,4’- trihydroxy isoflavone;
(c) obtaining at least one additive, at least one diluent, and at least excipients; and
(d) contacting the red algal extract, 6, 7, 4’- trihydroxy isoflavone, the at least one
additive, the at least one diluent, and the at least one excipient, to obtain said
composition.
9. A method to inhibit skin aging comprising:
(a) obtaining the composition as claimed in any of the claims 1-5; and
(b) contacting said composition with skin, wherein the composition inhibits skin aging.
10. The composition as claimed in any of the claims 1-6, wherein said composition
inhibits skin aging by inhibiting collagen degrading enzymes.