Patent ID: 12201717

DETAILED DESCRIPTION

The compositions of the present disclosure can comprise, consist essentially of, or consist of, the components of the present disclosure as well as other ingredients described herein. The term “comprising” as used herein is meant to include various optional, compatible components that can be used in the preservative systems and cosmetic compositions of the present disclosure without limiting the inclusion, use of, or cooperation with other ingredients, excipients, uses, or otherwise. The term “consisting essentially of” as used herein means that the composition or component may include additional ingredients, but only if the additional ingredients do not materially alter the basic and novel characteristics of the compositions or methods.

As used herein, the words “preferred,” “preferably,” and variants thereof refer to embodiments of the disclosure that afford certain benefits under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the disclosure.

Numerical ranges as used herein are intended to include every number and subset of numbers contained within that range, whether specifically disclosed or not. Further, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers within that range.

All percentages, parts, proportions, and ratios as used herein are by weight of the total composition, unless otherwise specified. All such weights as they pertain to listed ingredients are based on the active level.

All references to singular characteristics or limitations of the present disclosure shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary by the context in which the reference is made.

All publications, articles, papers, patents, patent publications, and other references cited herein are hereby incorporated in their entireties for all purposes to the extent consistent with the disclosure herein.

The term “dry skin-inducing water loss” as described herein refers to an amount of trans-epidermal water loss (TEWL) that causes skin to become dry, flaky, itchy and irritated, all signs of an improperly functioning stratum corneum/skin barrier.

The term “oxidative stress” as described herein refers to the disturbance in balance between reactive oxygen species (ROS), reactive nitrogen species (RNS) and/or other free radicals and antioxidants present in the skin caused by extrinsic and/or intrinsic factors. Extrinsic factors include, for example, exposure to UV radiation, pollution, and products containing harsh chemicals. Intrinsic factors include, for example, chronological aging, a person's genetic makeup, and other biological changes that occur from within the skin.

The term “free radicals” as described herein refers to those ROS and/or RNS that are formed when skin experiences oxidative stress caused by extrinsic and/or intrinsic factors.

The term “keratinocyte hyperproliferation” as described herein refers to uncontrolled epidermal cellular differentiation in support of skin barrier integrity.

The term “skin barrier homeostasis” as described herein refers to the ability of the stratum corneum to maintain balanced hydration levels in support of skin barrier integrity and functionality.

The present disclosure generally relates to compositions and methods for positively influencing (1) pro-inflammatory cellular signaling in response to oxidative stress, (2) hydration levels within the skin barrier, and (3) keratinocyte hyperproliferation in order to help support skin barrier homeostasis.

It has surprisingly been discovered by the inventors that a composition comprising a mixture of: (1) a first active blend of at least: (a) Jojoba oil/Macadamia seed oil esters; (b) squalene; (c) phytosteryl macadamiate; and (d) phytosterols; (2) a second active blend of at least: (e) EGF growth factor; (f) IGF-1 growth factor; (g) acidic FGF growth factor; (h) basic FGF growth factor; (i) VEGF growth factor; (j) vitamin B9; and (k) acetyl glutamine; and (3) a third active blend of at least: (l) an extract of lithothamnium calcareum; (m) pentylene glycol; (n) lactic acid; and (o) glycerin; (4) an emulsifier; and (5) a dermatologically acceptable carrier, when applied onto skin, downregulates pro-inflammatory cellular signaling in oxidatively-stressed skin, promotes water retention, and mitigates keratinocyte hyperproliferation to help support skin barrier homeostasis.

A first active blend comprised of: (a) Jojoba oil/Macadamia seed oil esters; (b) squalene; (c) Phytosteryl Macadamiate; and (d) phytosterols, is one of the three key constituents of the composition of the present disclosure. The active blend represents a lipid complex meant to mimic the surface lipid profile of a healthy, young adult and is used to help balance the stratum corneum's lipid network.

This first active blend is commercially available from International Flora Technologies, Ltd under the tradename L22®.

The first active blend may be employed in an amount of from about 0.25% to about 6% by weight, preferably in an amount of from about 0.25% to about 5% by weight, and more preferably in an amount of from about 0.25% to about 4% by weight, all weights based on the total weight of the composition.

A second active blend comprised of: (e) EGF growth factor; (f) IGF-1 growth factor; (g) acidic FGF growth factor; (h) basic FGF growth factor; (i) VEGF growth factor; (j) vitamin B9; and (k) acetyl glutamine, is another of the three key constituents of the composition. The active blend represents a combination of specific types of growth factors, amino acids and vitamins used for its efficacy in diminishing the appearance of wrinkles in human skin and other anti-aging benefits.

This second active blend is commercially available from Labio. Co., Ltd under the tradename BIO-PLACENTA®.

The second active blend may be employed in an amount of from about 0.1 to about 6% by weight, preferably in an amount of from about 0.1% to about 5% by weight, and more preferably in an amount of from about 0.25% to about 4% by weight, all weights based on the total weight of the composition.

A third active blend comprised of: (l) an extract of lithothamnium calcareum; (m) pentylene glycol; (n) lactic acid; and (o) glycerin, is the last of the three key constituents of the composition. The active blend represents a mixture of natural moisturizers used as a skin barrier enhancer due to its ability to provide hydration to the skin and defend against TEWL.

This third active blend is commercially available from Provital, S.A.U. under the tradename HYDRAFENCE™.

The third active blend may be employed in an amount of from about 0.25 to about 6% by weight, preferably in an amount of from about 0.25 to about 5% by weight, and more preferably in an amount of from about 0.25 to about 4% by weight, all weights based on the total weight of the composition.

According to one embodiment of the present disclosure, there is provided a composition intended for application onto human skin to help support skin barrier homeostasis, the composition comprising a mixture of: (1) a first active blend of at least: (a) Jojoba oil/Macadamia seed oil esters; (b) squalene; (c) phytosteryl macadamiate; and (d) phytosterols; (2) a second active blend of at least: (e) EGF growth factor; (f) IGF-1 growth factor; (g) acidic FGF growth factor; (h) basic FGF growth factor; (i) VEGF growth factor; (j) vitamin B9; and (k) acetyl glutamine; and (3) a third active blend of at least: (l) an extract of lithothamnium calcareum; (m) pentylene glycol; (n) lactic acid; and (o) glycerin; (4) an emulsifier; and (5) a dermatologically acceptable carrier, wherein (1)-(3) are employed in amounts sufficient to downregulate pro-inflammatory cellular signaling in oxidatively-stressed skin, thereby helping to support skin barrier homeostasis.

The inventors have surprisingly and unexpectedly discovered that a mixture of the three above-disclosed active blends, when applied onto skin, synergistically: (1) downregulates the inflammatory cascade triggered by oxidative stress in the stratum corneum, thereby mitigating damage experienced by healthy skin cells; (2) positively influences skin barrier-disrupting keratinocyte hyperproliferation; and (3) helps promote water retention by the skin, thereby helping to maintain skin barrier homeostasis. These synergies are only realized in the presence of the mixture. None of the three active blends, individually, yielded these synergistic skin barrier benefits.

The dermatologically acceptable carrier can encompass a wide variety of forms. In some cases, the solubility or dispersibility of the components in the composition may dictate the form and character of the carrier. Non-limiting examples include simple solutions (e.g., aqueous or anhydrous), dispersions, emulsions, and solid forms. In certain embodiments, the dermatologically acceptable carrier is in the form of an emulsion. An emulsion can be generally classified as having a continuous aqueous phase (e.g., oil-in-water and water-in-oil-in-water) or a continuous oil phase (e.g., water-in-oil or oil-in-water).

Any ingredient capable of emulsifying the composition may be employed as an emulsifier without departing from the spirit of the disclosure, so long as it is dermatologically acceptable. Examples thereof include, but are not limited to, glyceryl stearate, cetyl alcohol, sodium stearoyl lactylate, sorbitan olivate, cetearyl olivate, cetearyl alcohol, cetearyl glucoside, sodium cetearyl sulfate, and the like. It is also particularly preferred that the emulsifier be free of palm oil.

The compositions of the present disclosure may be made available to consumers in a wide variety of product forms that include, but are not limited to, solutions, suspensions, lotions, creams, gels, sprays, ointments, foams, and serums.

According to embodiments of the present disclosure, the compositions can also additionally comprise suitable optional ingredients as desired. For example, the composition can optionally include other active and/or inactive ingredients, provided they do not unacceptably alter the synergistic benefits of the skin care composition. The selection and precise amount of optional ingredients will be determined by those skilled in the art.

In yet another embodiment of the present disclosure, there is provided a method of downregulating pro-inflammatory cellular signaling in oxidatively stressed skin by applying one of the above-disclosed compositions onto the skin.

A further embodiment of the present disclosure provides for a method of positively influencing skin hydration to help support skin barrier homeostasis by applying one of the above-disclosed compositions onto oxidatively stressed skin.

In yet another embodiment of the present disclosure, there is provided a method of mitigating skin barrier-disrupting keratinocyte hyper-proliferation to support skin barrier integrity and functionality.

EXAMPLES

The following examples as set forth herein are intended for illustrative purposes only and are not intended to limit the scope of the disclosure in any way, as many variations thereof are possible without departing from the spirit and scope of the disclosure. In the examples, all concentrations are listed as weight percent, unless otherwise specified.

Example 1

Gene expression testing was performed in order to ascertain whether the composition of the present disclosure exhibited any synergistic properties as compared to its three key constituents, individually. Briefly, Reconstructed Human Epidermis (RHE) tissues were incubated with: (i) the first active blend of Jojoba oil/Macadamia seed oil esters+squalene+phytosteryl macadamiate+phytosterols; (ii) the second active blend of EGF growth factor+IGF-1 growth factor+acidic FGF growth factor+basic FGF growth factor+VEGF growth factor+vitamin B9+acetyl glutamine; (iii) the third active blend of an extract of lithothamnium calcareum+pentylene glycol+lactic acid+glycerin; and (iv) a combination of (i)-(iii). Following the incubation, RNA was extracted from the tissue, and real time quantitative PCR was performed. The efficiency ΔΔCt method was used for quantification of results, after the normalization of gene expression to conventional housekeeping genes. Genes were considered differentially expressed if there was a ±1.5 average Fold Change that was statistically significant. Statistical significance was based on the p value being ≤0.05.

While tissue treated with each of the individual active blends showed bioactivity, it was surprisingly discovered that tissues treated with a composition containing all three active blends exhibited “synergistic” bioactivity in its ability to down-regulate pro-inflammatory cellular signaling, support skin barrier homeostasis, and positively influence keratinocyte proliferation, as is seen in Table 1, below.

TABLE 1AvgAvgAvgAvgFoldFoldFoldFoldChangeP-valChangeP-valChangeP-valChangeP-valAmountforforforforforforforforIngredient(wt %)CX3CR1CX3CR1DDX58DDX58CDK4CDK4SHROOM3SHROOM3First active3.0%1.000.55−1.170.66−1.330.351.050.76blendSecond0.5%−1.060.991.060.871.040.55−1.710.12active blendThird active0.25%−1.521.2−1.610.25−1.280.31−1.200.36blendComplex 13.75%−2.120.04−2.020.04−1.910.03−2.030.03AvgAvgAvgAvgFoldFoldFoldFoldChangeP-valChangeP-valChangeP-valChangeP-valAmountforforforforforforforforIngredient(wt %)TAF15TAF15LRRC8ALRRC8ACLCN6CLCN6IFI16IFI16First blend3.0%−1.290.521.490.0561.070.89−1.500.06Second0.5%1.090.38−1.560.008−1.030.091.270.12blendThird blend0.25%−1.090.641.530.0061.010.96−1.550.10Complex 13.75%−1.730.011.710.002−1.770.001−1.980.003

The data in Table 1 shows that tissue treated with the composition of the present disclosure induced a statistically significant synergistic down-regulation of pro-inflammatory cell signaling. More particularly, with regards to genes CX3CR1, DDX58, and SHROOM3, only the mixture exhibited statistically significant (i.e., P-val≤0.05) and synergistic (i.e., Avg Fold Change of greater than or equal to ±1.5) differential expression of these genes, as compared to each individual active blend.

In addition, it was also surprisingly discovered that genes TAF15, LRRC8A, CLDN-6, CDK4, and IFI16 also showed statistically significant synergistic differential expression only in the presence of the composition and not in the presence of the individual blends. Genes TAF15, CLDN-6, and CDK4 covering keratinocyte hyperproliferation and LRRC8A covering cellular osmotic (water retention) changes, by virtue of their being synergistically expressed by the inventive mixture, evidence the ability of the mixture to synergistically enhance homeostasis within the skin. In addition, gene IFI16 (linked to inflammatory cytokine expression in cells) was also positively influenced to a statistically significant degree by the mixture, evidencing a synergistic improvement in skin barrier integrity and protective functionality.

Example 2

A second composition, also in accordance with the present disclosure, was evaluated for the presence of synergistic skin barrier-supporting properties.

TABLE 2AvgAvgAvgAvgFoldP-valFoldFoldFoldChangeforChangeP-valChangeP-valChangeP-valAmountfor IFIIFIforforforforforforIngredient(wt %)1616CDK4CDK4LOXLOXIL37IL37First active0.25%1.320.181.020.401.120.451.740.23blendSecond active0.1%−1.040.88−1.220.17−1.030.801.290.52blendThird active0.25%−1.550.101.280.31−1.300.112.440.02blendComplex 20.6%−1.550.101.050.60−1.680.0051.970.02

Gene CDK4 relating to epidermal cellular differentiation evidences the ability of the composition to synergistically help support skin barrier integrity by positively influencing keratinocyte hyperproliferation. In addition, the genes LOX, IL37, and IFI16 relating to inflammatory cytokine expression in cells were also positively influenced to a statistically significant degree by the mixture, evidencing a synergistic improvement in managing the inflammatory cascade leading to enhanced skin barrier integrity and functionality.

Example 3

A skincare composition in accordance with the present disclosure was prepared comprising the ingredients shown in Table 3, below:

TABLE 3Ingredient (by tradename)Wt. %Bio-Placenta1.00%Hydrafence0.25%L223.00%Carrier Vehicleq.s. 100%

The composition of Example 3 was clinically tested to evaluate its ability to help skin maintain water homeostasis (i.e., constant hydration levels) to help maintain skin barrier integrity. Thirty-two females aged 41-73 were asked to apply the composition under normal use conditions onto an area of skin surrounding their eye orbital twice a day, once in the morning and once at night. After taking an initial baseline reading, skin health and appearance data were collected 1 hour after initial application on day 0, then on day 7, and day 14.

Hydration data was collected using a Corneometer® which measures skin capacitance based on humidity levels in the skin. The results obtained are seen in Table 4, below.

TABLE 4Mean %% of subjectsVisitimprovementimprovedHour 1 (±10 minutes)18.0%96.9%Day 74.6%56.3%Day 2825.8%90.6%

The results obtained in Table 4 evidence the ability of the composition of Example 3 to support skin barrier integrity via significantly improved water homeostasis within the skin. Approximately 97% of test subjects experienced an 18.0% improvement in hydration after 1 hour post application, with approximately 91% of test subjects experiencing a 25.8% improvement after week 4.

Next, the composition of Example 3 was evaluated using Expert Clinical Grading Scores calculated using the Wilcoxon signed rank test with a null hypothesis that mean change from baseline is zero, to evaluate its effect on skin homeostasis, as is seen in Table 5, below.

TABLE 5Mean %% of subjectParameterVisitimprovementimprovedUnder Eye DarkHour 1 (±10 mins)4.312.5CirclesDay 714.962.5Day 2821.375.0Fine Lines in CrowsHour 1 (±10 mins)2.16.3Feet AreaDay 714.962.5Day 2821.371.9Wrinkles in CrowsHour 1 (±10 mins)0.00.0Feet AreaDay 76.518.8Day 2815.256.3Radiance of Skin inHour 1 (±10 mins)14.853.1Eye Area treatedDay 724.168.8Day 2831.587.5Under Eye PuffinessHour 1 (±10 mins)5.928.1Day 715.779.1Day 2831.496.9Sagging in Eye AreaHour 1 (±10 mins)2.56.3Day 77.525.0Day 2820.059.4

Expert Clinical Grading data showed that the composition of Example 3 significantly improved the parameters identified in Table 6 after weeks 1 and 4 evidencing the ability of the inventive composition to help support skin barrier integrity and homeostasis.

The test subjects were then asked to assess, qualitatively, their impression of the composition's efficacy on their skin. 10 out of 15 had a positive impression after the initial application; 15 out of 15 had a positive impression after 7 days; and 15 out of 15 had a positive impression after 14 days of use.

Example 4

A skincare composition in accordance with the present disclosure was prepared comprising the ingredients shown in Table 6:

TABLE 6Ingredient (by tradename)Wt. %Bio-Placenta0.10%Hydrafence0.25%L220.25%Carrier Vehicleq.s. 100%

The composition of Example 4 was clinically tested to evaluate its ability to address skin barrier-disrupting keratinocyte hyperproliferation in the form of keratosis pilaris. Twenty-one individuals aged 18-61 were asked to apply the composition, twice a day, onto an area of their skin where the condition keratosis pilaris was present. After taking an initial baseline reading, data was collected 15 minutes (+5 mins) after initial application on day 0, then on day 7, and day 14.

Next, the composition of Example 4 was evaluated using Expert Clinical Grading Scores calculated using the Wilcoxon signed rank test with a null hypothesis that mean change from baseline is zero, to evaluate its effect on keratinocyte hyperproliferation, as is seen in Table 7, below.

TABLE 7Mean %% of subjectParameterVisitimprovementimprovedKeratosis Pilaris15 mins (±5 mins)0.00.0Day 7 (±1 day)0.04.8Day 14 (±1 day)8.628.6Skin Texture15 mins (±5 mins)5.019.0Day 7 (±1 day)17.561.9Day 14 (±1 day)25.076.2Skin Flaking15 mins (±5 mins)68.881.0Day 7 (±1 day)87.585.7Day 14 (±1 day)87.585.7

Expert Clinical Grading data showed that the composition of Example 3 significantly improved all of the parameters identified in Table 8 after day 7 (+1 day) and day 14 (+1 day), evidencing the ability of the inventive composition to positively influence keratinocyte hyperproliferation and thereby support skin barrier integrity.

The test subjects were then asked to assess, qualitatively, their impression of the composition's efficacy on their skin. 9 out of 15 had a positive impression after the initial application; 12 out of 15 had a positive impression after 7 days; and 13 out of 15 had a positive impression after 14 days of use.

Example 5

A skincare composition in accordance with the present disclosure was prepared comprising the ingredients shown in Table 8, below:

TABLE 8Ingredient (by tradename)Wt. %Bio-Placenta3.00%Hydrafence3.00%L223.00%Carrier Vehicleq.s. 100%

The composition of Example 5 was clinically tested to evaluate its ability to positively influence the appearance of fine lines, wrinkles, skin elasticity, and skin plumpness. Thirty-three females aged 35-70 were asked to apply the composition under normal use conditions onto their face twice a day, once in the morning and once at night. After taking an initial baseline reading, skin health and appearance data were collected 15 minutes after initial application on day 0, then on day 7, and finally on day 28.

Hydration data was collected using a CORNEOMETER® which measures skin capacitance based on humidity levels in the skin. The results obtained are seen in Table 9, below.

TABLE 9VisitMean % improvement% of subjects improved15 minutes32.0%93.9%Day 7−13.8%45.5%Day 2815.1%75.8%

The results obtained in Table 9 evidence the ability of the composition of Example 5 to support skin barrier integrity via significantly improved water homeostasis within the skin. Approximately 94% of test subjects experienced an 32.0% improvement in hydration after 15 minutes post application, with approximately 76% of test subjects experiencing a 15.1% improvement at day 28.

Next, the composition of Example 5 was evaluated using Expert Clinical Grading Scores calculated using the Wilcoxon signed rank test with a null hypothesis that mean change from baseline is zero, to evaluate its effect on skin homeostasis, as is seen in Table 10, below.

TABLE 10Mean %% of subjectParameterVisitimprovementimprovedSkin Plumpness15 min.Not Stat. SignificantN/ADay 7−6.053.3Day 28−8.570.0Fine Lines15 min.−6.763.3Day 7−11.686.7Day 28−15.886.7Wrinkles15 min.N/AN/ADay 7N/AN/ADay 28−5.943.3Skin Firmness15 min.N/AN/ADay 7N/AN/ADay 28−2.726.7Skin Elasticity15 min.N/AN/ADay 7N/AN/ADay 28−2.423.3Overall Appearance15 min.N/AN/Aof Skin (Healthy)Day 7N/AN/ADay 28−5.660.0

Expert Clinical Grading data showed that the composition of Example 5 improved, in a statistically significant way, the parameters identified in Table 10 after evidencing the ability of the inventive composition to help support skin barrier integrity and homeostasis.

The test subjects were then asked to assess, qualitatively, their impression of the composition's efficacy on their skin in terms of its appearance and overall health. 93.9% had a positive impression 15 minutes after the initial application; and 90.9 had a positive impression after 7 and 28 days of use.

Example 6

A skincare composition in accordance with the present disclosure was prepared comprising the ingredients shown in Table 11, below:

TABLE 11Ingredient (by tradename)Wt. %Bio-Placenta4.00%Hydrafence4.00%L224.00%Carrier Vehicleq.s. 100%

The composition of Example 6 was clinically tested to evaluate its ability to positively influence skin hydration levels and trans-epidermal water loss (TEWL). Thirty-one females aged 21-64 were asked to apply the composition under normal use conditions onto their face twice a day, once in the morning and once at night. After taking an initial baseline reading, skin health and appearance data were collected 30 minutes (+/−5 min.) after initial application on day 0 and on day 14.

Hydration data was collected using a CORNEOMETER® which measures skin capacitance based on humidity levels in the skin. The results obtained are seen in Table 12 below.

TABLE 12VisitMean % improvement% of subjects improved30 minutes (+/− 5 min.)22.7%93.5%Day 1421.7%83.9%

The results obtained in Table 12 evidence the ability of the composition of Example 6 to support skin barrier integrity as is evidenced by the significant improvement in hydration levels within the skin. Approximately 94% of test subjects experienced a 22.7% improvement in hydration after 30 minutes post application, with approximately 84% of test subjects experiencing a 21.7% improvement at day 14.

Trans-epidermal water loss (TEWL) data was collected using a VAPOMETER® which measures relative humidity levels in the skin. The results obtained are seen in Table 13, below.

TABLE 13VisitMean % improvement% of subjects improved30 minutes (+/− 5 min.)13.9%93.5%Day 1417.9%90.3%

The results obtained in Table 13 evidence the ability of the composition of Example 5 to support skin barrier integrity as is evidenced by reduced TEWL by the skin evidencing an improvement in skin barrier function. Approximately 94% of test subjects experienced a 13.9% improvement in reduced TEWL after 30 minutes post application, with approximately 90% of test subjects experiencing a 17.9% improvement at day 14.

The test subjects were then asked to assess, qualitatively, their impression of the composition's efficacy on their skin in terms of its appearance and overall health. 100% had a positive impression of their skin 30 minutes after the initial application, and after 14 days of use.

By providing skin care compositions and methods according to the present disclosure, the damage done to healthy skin cells when the body responds to oxidative stress in the stratum corneum can be synergistically mitigated in a statistically significant way. The downregulation of pro-inflammatory cellular signaling in response to oxidative stress mitigates the damage done to healthy skin cells during the body's inflammatory response to oxidative stress.

In addition, the findings relating to both keratinocyte proliferation (epidermal cellular differentiation) and water homeostasis (cellular osmotic changes) validate the invention's ability to synergistically support skin barrier homeostasis/equilibrium and, consequently, skin barrier health and functionality.

The skilled artisan will recognize the interchangeability of various components of different embodiments described. Besides the variations described, other known equivalents for each feature can be mixed and matched by one of ordinary skill in this art to arrive at a preservative system under principles of the present disclosure. Therefore, the embodiments described may be adapted to preservative systems for other types of compositions and for other uses than those described and having other components than those described.

Although compositions and methods have been disclosed in certain preferred embodiments and examples, it nevertheless will be understood by those skilled in the art that the present disclosure extends beyond the disclosed embodiments to other alternative embodiments and/or uses of the disclosure and obvious modifications and equivalents. It is intended that the scope of the present disclosure should not be limited by the disclosed embodiments described above but should be determined only by a fair reading of the claims that follow.