Skin care composition utilizing the activity of sunflower sprouts subjected to metabolomics-guided bioprocess operation

Abstract

The present disclosure relates to a skin care composition comprising sunflower sprout extract for treating skin conditions. More specifically, the novel aspect is a metabolomics-guided bioprocess engineered sunflower sprout extract obtained from sunflower seeds germinated in a controlled environment, in which free fatty acids are degraded to create intracellular energy important for vitality. By incorporating the metabolomics-guided bioprocess engineered sunflower sprout extract as an active ingredient in a skin care composition that acts via epigenetic pathways, as a result, human gene expression, as well as protein and lipid production, are enhanced in correlation with an anti-aging effect. The skin care composition comprises an enhanced extract obtained from plant seeds germinated in an environment containing an effective amount of sucrose and an improved oxygen level, and a cosmetically acceptable carrier.

Description

Technical Field

[0001] [Cross - Reference to Related Applications] This application claims priority to U.S. Application No. 17 / 841,038, filed on June 15, 2022, entitled "Skincare Composition Utilizing a Metabolomics Guided, Bioprocess Engineered Sunflower Sprout Active", which is hereby incorporated by reference in its entirety.

[0002] The novel aspects of the present disclosure relate to a skincare composition comprising a sunflower sprout extract for treating skin conditions. More specifically, the novel aspects are directed to a metabolomics guided, bioprocess engineered sunflower sprout extract obtained from sunflower seeds germinated in a controlled environment in which free fatty acids are degraded to create intracellular energy important for vitality. Incorporating the metabolomics guided, bioprocess engineered sunflower sprout extract as an active ingredient in a skincare composition that acts via epigenetic pathways results in enhanced human gene expression and production of proteins and lipids, which correlates with an anti - aging effect.

Background Art

[0003] Human skin is a complex multi-layered tissue from the perspectives of its structure and biochemistry. Skin ages over time, and its visible appearance and texture change. Such changes are the result of both exogenous and endogenous factors. Examples of external factors that cause skin aging include nutrition, as well as exposure to sunlight, pollution, and high-energy visible light from electronic devices. Topical formulations have been used to minimize or reverse these changes. As research on human skin progresses, new treatments for improving skin aging are being developed.

Summary of the Invention

[0004] The novel aspect of the present disclosure is directed to a skin care formulation comprising a metabolomics-derived bioprocess-engineered sunflower sprout extract mixed with a cosmetically acceptable carrier. The metabolomics-derived bioprocess-engineered sunflower sprout extract is obtained from plant seeds germinated in an environment enhanced with an effective amount of sucrose and enriched with increased levels of oxygen.

[0005] The novel aspect of the present disclosure includes a method for improving the effects of skin aging, the method comprising applying a skin care composition to human skin, the skin care composition comprising a cosmetically acceptable carrier and a mixture with a metabolomics-derived bioprocess-engineered sunflower sprout extract obtained from plant seeds germinated in an environment enhanced with an effective amount of sucrose and enriched with increased levels of oxygen.

[0006] New aspects of the present disclosure also relate to a method for preparing a skin care formulation for use in improving the effects of aging, the method comprising obtaining a metabolomics-induced bioprocess-engineered sprout extract formed from plant seeds germinated in an environment enhanced by the addition of sucrose and fortified by increased oxygen levels, and mixing the metabolomics-induced bioprocess-engineered sprout extract with a cosmetically acceptable carrier.

[0007] Other aspects, embodiments, and features of the invention will become apparent from the following detailed description of the invention when considered in conjunction with the accompanying drawings. In the drawings, components that are the same or substantially similar, as illustrated in the various figures, are represented by a single number or notation. For clarity, not all components are shown in all figures. Also, not all components of each embodiment of the invention are shown when illustration is not necessary for those of ordinary skill in the art to understand the invention.

Brief Description of the Drawings

[0008] The novel features that are characteristic of the invention are set forth in the appended claims. However, like the invention itself, its preferred usage forms, further objects, and advantages will be best understood by reference to the detailed description of the following exemplary embodiments when read in conjunction with the accompanying drawings.

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Mode for Carrying Out the Invention

[0009] [Detailed Description] With aging, the energy production of human skin cells decreases, and the amount of antioxidants also decreases. Furthermore, the level of matrix metalloproteinase (MMP), an enzyme that degrades extracellular matrix (ECM) proteins, increases, leading to the loss of elasticity and firmness. Examples of ECM proteins include collagen and elastin. Various studies, such as gene array analysis and in vitro experiments, have shown that certain skin care components have beneficial effects on the structure and function of the skin. These components can beneficially regulate the expression of genes that are particularly correlated with minimizing the aging process through epigenetic effects. For example, certain components can upregulate the expression of genes involved in cellular energy production. Alternatively, certain components can downregulate the expression of genes related to the production of MMPs that degrade valuable ECM proteins. This type of skin care active ingredient has been the focus of ongoing research.

[0010] Consumers are increasingly preferring skin care compositions having activities derived from natural, non-genetically modified, plant-based ingredients. Sunflower plant and seed-based extracts have been studied as sources of cosmetic active ingredients because sunflower plants and seeds are widely available, can be grown in many different types of soil, and exhibit natural allelopathic potential (i.e., the ability to inhibit or stimulate biological interactions by the release of biochemicals). Conventionally, sunflower-based extracts used in skin care compositions have utilized sunflower seed extracts obtained from sunflower seeds harvested under normal growing conditions, or sunflower extracts obtained from hydroponically grown sunflower shoots that are soaked in water for two days and then transferred to a rotating vessel for a four-day incubation period.

[0011] These conventional skin care compositions containing the above sunflower extract are an incomplete solution that cannot utilize all plant gene expression regions available through metabolomics-guided bioprocess engineering. Metabolomics-guided bioprocess engineering changes the expression of plant genes without changing the genes themselves, resulting in the production of metabolites that are not normally produced under normal growth conditions or are produced only in limited amounts. A metabolomics-guided bioprocess method is described that optimizes the germination process of sunflower seeds by adding sugar and oxygen and enhances the production of beneficial metabolites in germinated sunflower sprouts, and the germinated sunflower sprouts contain the presence of L-carnitine (LCAR), acetyl-L-carnitine (ALCAR), L-α-phosphatidylcholine, galactose phosphate, glucose phosphate, adenosine triphosphate, inositol, triglyceride, docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA).

[0012] Although there is evidence suggesting that the intake of LCAR and ALCAR metabolites isolated from sunflower seeds germinated by the metabolomics-induced bioprocess method is beneficial for the treatment of neuropathy, the available literature does not describe the effect of LCAR and ALCAR, together with the whole-region metabolites of sunflower sprout extract formed by bioprocess manipulation, on reversing the aging effect on human skin. For this purpose, a novel aspect of the present disclosure is an enhanced sunflower sprout extract formed by a metabolomics-induced bioprocess engineering method of germinating sunflower seeds that produce metabolites important for skin health and anti-aging purposes, the enhanced sunflower sprout extract being absent or present in only ineffective amounts in conventional germinated sunflower seeds. For example, in an enhanced extract obtained from sunflower sprouts germinated in an environment reinforced and enhanced with an oxygen concentration and an effective concentration of sucrose solution, the amounts of LCAR and ALCAR, metabolites that are not produced or are produced in only very limited amounts in normal sprouts, are increased by up to 1000 times. For use as a skin care composition, incorporating the enhanced sunflower sprout extract into a cosmetically acceptable carrier has been shown to enhance cell energy, enhance skin glow and suppleness, optimize the epidermal barrier, enhance skin elasticity, and promote long-term skin health through an epigenetic process.

[0013] The various embodiments disclosed herein describe a metabolomics-induced bioprocess-engineered sunflower sprout extract obtained from sunflower seeds, i.e., an enhanced extract, for use as an active ingredient in a skin care composition for improving the effects of aging, provided that other plant seeds can be substituted provided they have lipid storage substances stored in their endosperm. Sunflower seeds store lipid storage substances in their cotyledons.

[0014] Figure 1 is a flowchart of steps for forming a bio-processed sunflower sprout extract. The steps of this process in flowchart 100 are described in further detail in U.S. Patent No. 7,632,662, which is hereby incorporated by reference in its entirety, but is summarized here for convenience of reference.

[0015] The metabolomics-guided bioprocess method manipulates the metabolic flux of germinating seeds to enhance oxygen availability, and recently, provides a sterile environment free of contamination such as viruses or other toxic organisms, changes hydrocarbon consumption, and feedback inhibits gluconeogenesis. The shifts resulting from the fatty acid metabolism, phospholipid metabolism, and high-energy metabolism of sunflower seeds increase beneficial metabolites such as LCAR and ALCAR, and change the germination environment to produce long-chain fatty acid esters of carnitine such as polyunsaturated fatty acid esters, such as eicosapentaenoyl-L-carnitine and docosahexaenyl-L-carnitine. In a non-limiting embodiment, the method manipulates the metabolic flux by providing a sucrose-controlled and oxygen-controlled environment.

[0016] Flowchart 100 begins in step 102 by sterilizing sunflower seeds. In one embodiment, the sunflower seeds are sterilized to remove contaminants by application of a conventional sterilizing solution. The sterilization step may include rinsing with deionized water before and / or after sterilization.

[0017] In step 104, the sterilized sunflower seeds are cultured in the presence of an effective amount of sucrose solution and 40% oxygen. In a non-limiting embodiment, the effective amount of sucrose solution for sunflower seeds is 1% - 2% w / v. In a specific embodiment, the effective amount of sucrose solution for sunflower seeds is 2% w / v. For example, to maintain 40% oxygen, the headspace of the container can be purged with oxygen, and sunflower seeds can be placed in a container containing a culture medium formed from 2% w / v sucrose in deionized water. The sunflower seeds are cultured in the container for at least 60 hours, or until at least the hypocotyl elongates, to form metabolomics-induced bioprocess-engineered sunflower sprouts, i.e., enhanced sprouts. The term "sunflower sprouts" means sunflower seeds cultured in the presence of an effective amount of sucrose solution and 40% oxygen for at least 60 hours, or until at least the hypocotyl elongates, when referring to sunflower seeds that are the subject of metabolomics-induced bioprocess engineering.

[0018] In step 106, optionally, the sunflower sprouts are washed. The washing water can be deionized water or distilled water. In step 108, the sunflower sprouts are ground into a paste to form a sunflower sprout extract. In one embodiment, the sunflower seeds are ground into a paste at liquid nitrogen temperature, for example, about -195°C or about -320°F.

[0019] To test the effect of the metabolomics-induced bioprocess-engineered sunflower sprout extract on epigenetics and gene expression, skin samples collected from the abdomen of female donors and divided into three separate samples (n = 3) were cultured for 24 hours in a culture medium containing 100% (w / w) concentration of enhanced extract and a water control, and 20,000 genes were analyzed to examine the potential anti-aging effect on human skin. Only genes showing a large fold change and a p-value of less than 0.05 were considered to have a difference in expression. The results are shown in Tables 1 and 2 below.

[0020]

Table 1

[0021]

Table 2

[0022] The genes listed in Table 1 are involved in the repair and function of the epidermal barrier, in addition to cell respiration and energy production. The genes listed in Table 2 are involved in inflammation. Therefore, down-regulation of these genes suppresses inflammation and pushes the skin into a healthier state. Considering together with the up-regulated genes listed in Table 1, the metabolomics-induced bioprocess-engineered sunflower sprout extract affected 12 genes targeting skin barrier repair, melanogenesis, cellular energy, inflammation and extracellular matrix production. In contrast, the conventional extract formed from sunflower shoot extract cultured with human keratinocytes showed two highly up-regulated genes, filaggrin and γ-cystathionase (formerly known as γ-glutamylcysteine synthetase), and three highly down-regulated genes, MMP-1, MMP-3, and MMP-9. Therefore, the sunflower shoot extract affected only 5 genes targeting skin barrier repair and antioxidant production.

[0023] To demonstrate the effects of metabolites present in conventional sunflower shoot extracts, the gene expression of dermal fibroblasts was tested in comparison to a control. Human skin fibroblasts were first treated with H2O2 to mimic the effects of aging. The control group was cultured in a culture medium without sunflower shoot extract, and the experimental group was cultured in a culture medium containing sunflower shoot extract. In the control group, downregulation of specific genes such as CCN13, IGFBP2, PDGFR, CCNE1, P53, ACAN, COL3A1, and TMC was observed, and upregulation of a gene that promotes ECM destruction, namely MMP-3, was seen. These results are consistent with aging. When cultured in the presence of sunflower shoot extract, the genes downregulated by H2O2, namely CCN13, IGFBP2, PDGFR, CCNE1, P53, ACAN, COL3A1, and TMC, were upregulated in the presence of sunflower shoot extract, and the gene upregulated by H2O2, namely MMP-3, was downregulated by sunflower shoot extract. To compare the effects of sunflower sprout extract, skin collected from the abdomen of female donors was also treated with H2O2 and then cultured in the presence of sunflower sprout extract, and the resulting gene expression was analyzed. Only genes showing a large fold change and a p-value of less than 0.05 were considered to have a difference in expression. Comparing the above table with the gene list, no similarity was found in the genes showing changes in gene expression, leading to the conclusion that different metabolic products with different anti-aging effects are formed by different processes.

[0024] In one embodiment, the metabolomics-guided bioprocessed sunflower sprout extract can be applied directly to the skin at a concentration of 100%. In another embodiment, the metabolomics-guided bioprocessed sunflower sprout extract can be mixed with a cosmetically acceptable carrier for application to the skin at a concentration of less than 100% in a skin care composition. For example, 0.25 to 25% of the skin care composition can be the metabolomics-guided bioprocessed sunflower sprout extract. In another example, 25 to 50% of the skin care composition can be the metabolomics-guided bioprocessed sunflower sprout extract. In yet another example, more than 50% of the skin care composition can be the metabolomics-guided bioprocessed sunflower sprout extract. An exemplary method of forming the skin care composition is described in FIG. 2 below.

[0025] FIG. 2 is a flowchart of steps for preparing a skin care composition using a metabolomics-guided bioprocessed sunflower sprout extract according to an exemplary embodiment. Flowchart 200 begins at step 202 by obtaining a metabolomics-guided bioprocessed sunflower sprout extract formed by the process of flowchart 100. At step 204, the metabolomics-guided bioprocessed sunflower sprout extract is optionally mixed with a cosmetically acceptable carrier. The cosmetically acceptable carrier can be an aqueous or anhydrous oily gel, or an oil-in-water, water-in-oil, silicone-in-water, or water-free emulsion.

[0026] The skin care composition formed by flowchart 200 of FIG. 2 can be used to improve the effects of aging on human skin according to an exemplary embodiment. This skin care composition can include a cosmetically acceptable carrier mixed with sunflower sprout extract obtained from sunflower seeds germinated in an environment enhanced by oxygen and an optimal concentration of sucrose solution, and the skin care composition can be applied to human skin at an application frequency for a specific skin condition. The skin care composition can also be applied at an application frequency based on desired results such as improvement of cell energy, skin glow, epidermal barrier repair, skin softness, increased elasticity and firmness, and long-term skin health.

[0027] To confirm the effectiveness of the new skin care formulation, a pilot test was conducted by instructing a small number of female and male participants to apply the new skin care formulation to their skin for two weeks. The participants were asked to evaluate the perceived effectiveness of the skin care formulation in six categories and provide an evaluation score from 0 to 5 corresponding to a range of 0% improvement (i.e., 0) to 100% improvement (i.e., 5) in 20% increments. The results of the perceived effectiveness are shown in FIGS. 3-8 below. The participants were also asked to evaluate the appeal of the skin care formulation in four different categories and provide an evaluation score from 0 to 5 corresponding to a range of no opinion (i.e., 0) to 100% positive opinion (i.e., 5) in 20% increments. The evaluation results of the appeal of the skin care product are shown in FIGS. 9-12 below.

[0028] FIG. 3 is a chart showing the perceived effect of the new skin care formulation regarding the reduction of fine lines. From chart 300, it can be seen that all participants perceived that the new skin care formulation reduced the appearance of fine lines. The majority of the participants gave a score of 4 out of 5.

[0029] Figure 4 is a chart representing the perceived effect of the new skin care formulation on the reduction of deep wrinkles. From chart 400, it can be seen that all participants perceived that the new skin care formulation reduced the appearance of deep wrinkles. Half of the participants gave it a score of 3 out of 5.

[0030] Figure 5 is a chart representing the perceived effect of the new skin care formulation on the improvement of skin tightening. From chart 500, it can be seen that all participants perceived that the new skin care formulation improved skin tightening. The majority of the participants gave it a score of 4 out of 5.

[0031] Figure 6 is a chart representing the perceived effect of the new skin care formulation on the improvement of skin brightness. From chart 600, it can be seen that all participants perceived that the new skin care formulation improved skin brightness. The majority of the participants gave it a score of 4 out of 5.

[0032] Figure 7 is a chart representing the perceived effect of the new skin care formulation on the improvement of skin texture. From chart 700, it can be seen that all participants perceived that the new skin care formulation improved skin texture. The majority of the participants gave it a score of 4 out of 5.

[0033] Figure 8 is a chart representing the perceived effect of the new skin care formulation on the improvement of skin uniformity. From chart 800, it can be seen that all participants perceived that the new skin care formulation improved skin uniformity. The majority of the participants gave it a score of 4 out of 5.

[0034] The following table represents the statistical analysis of the results of the experimental studies presented in Figures 3 - 8.

[0035] Table 3. Skin Care Data Analysis

Table 3

[0036] Figure 9 is a chart showing the appealing power of the color of a skin care preparation according to an exemplary embodiment. From chart 900, it can be seen that all participants recognize the color of the new skin care preparation as appealing. The majority of participants gave it 4 out of 5 points.

[0037] Figure 10 is a chart showing the appealing power of the fragrance of a skin care preparation according to an exemplary embodiment. From chart 1000, it can be seen that all participants recognize the fragrance of the new skin care preparation as appealing. The majority of participants gave it 4 out of 5 points.

[0038] Figure 11 is a chart showing the appealing power of the viscosity of a skin care preparation according to an exemplary embodiment. From chart 1100, it can be seen that all participants recognize the viscosity of the new skin care preparation as appealing. The majority of participants gave it 4 out of 5 points.

[0039] Figure 12 is a chart showing the appealing power of the absorbency of a skin care preparation according to an exemplary embodiment. From chart 1200, it can be seen that all participants recognize the absorbency of the new skin care preparation as appealing. The majority of participants gave it 5 out of 5 points.

[0040] Although embodiments of the present invention have been described with reference to several elements, any element described herein is exemplary and can be appropriately omitted, substituted, added, combined, or rearranged to form new embodiments. Those skilled in the art will recognize that such additional embodiments are effectively disclosed herein upon reading this specification. For example, when the present disclosure describes features, structures, sizes, shapes, arrangements, or compositions of a process for manufacturing or using an element, or an element or combination of elements, the features, structures, sizes, shapes, arrangements, or compositions can be incorporated into any other element or combination of elements described herein, or into a process for manufacturing or using an element or combination of elements to provide additional embodiments.

[0041] Furthermore, in this specification, when an embodiment is described as including an element or group of elements, additional embodiments can consist essentially of or consist of that element or group of elements. Also, although the open-ended term "comprises" is generally used in this specification, additional embodiments can be formed by replacing it with the terms "consist essentially of" or "consisting of".

[0042] The invention has been shown and described with reference to particular preferred embodiments, but it will be understood by those skilled in the art that various changes in form and detail can be made without departing from the spirit and scope of the invention. The inventors expect those skilled in the art to adopt such modifications as appropriate, and the inventors intend the invention to be practiced in a manner other than specifically described herein. Accordingly, the invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Further, any combination of the above-described elements in all possible variations is included in the invention unless otherwise specifically indicated herein or clearly inconsistent with the context.

Claims

1. A skincare composition for human use, An enhanced extract derived from plant seed sprouts germinated in an environment enhanced and enriched with oxygen and an effective concentration of sucrose solution, wherein the plant seed has lipid stores not stored in the endosperm, and the enhanced extract, A cosmetically acceptable carrier is mixed with the enhanced extract. A skincare composition for human use, including [the specified ingredient].

2. The human skincare composition according to claim 1, wherein the plant seed is a sunflower seed.

3. The human skincare composition according to claim 2, wherein the effective concentration of the sucrose solution is 2% w / v, and the amount of oxygen enhanced is 40% oxygen in the headspace of the sunflower seed.

4. The skincare composition according to claim 2, wherein the sunflower sprout extract comprises at least one of L-carnitine (LCAR), acetyl-L-carnitine (ALCAR), L-α-phosphatidylcholine, galactose phosphate, glucose phosphate, adenosine triphosphate, inositol, triglycerides, docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA).

5. The skincare composition according to claim 1, wherein the plant seeds exhibit gene expression modified from non-recombinant genes.

6. The skincare composition according to claim 5, wherein the modified gene expression yields an amount of LCAR about 1,000 times greater than the amount of LCAR yielded by the unmodified gene expression of the non-recombined gene.

7. The skincare composition according to claim 5, wherein the modified gene expression yields an amount of ALCAR about 1,000 times greater than the amount of ALCAR yielded by the unmodified gene expression of the non-recombined gene.

8. A method to improve the effects of aging on human skin, A method comprising applying a skincare composition to human skin, wherein the skincare composition comprises a cosmetically acceptable carrier mixed with an enhanced sprout extract obtained from plant seeds germinated in an environment enhanced and reinforced with oxygen and an effective concentration of sucrose solution, the plant seeds having lipid reserves not stored in the endosperm.

9. The method according to claim 8, wherein the plant seed is a sunflower seed.

10. The method according to claim 9, wherein the effective concentration of the sucrose solution is 2% w / v, and the amount of oxygen enhanced is 40% oxygen in the headspace of the plant seed.

11. The method according to claim 9, wherein the enhanced extract comprises at least one of L-carnitine (LCAR), acetyl-L-carnitine (ALCAR), L-α-phosphatidylcholine, galactose phosphate, glucose phosphate, adenosine triphosphate, inositol, triglycerides, docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA).

12. The method according to claim 8, wherein the plant seeds exhibit modified gene expression from non-recombinant genes.

13. The method according to claim 12, wherein the modified gene expression yields an amount of LCAR about 1000 times greater than the amount of LCAR yielded by the unmodified expression of a non-recombinant gene.

14. The method according to claim 12, wherein the modified gene expression yields an amount of ALCAR about 1000 times greater than the amount of ALCAR yielded by the unmodified expression of a non-recombinant gene.

15. The method according to claim 8, wherein the skincare composition is applied to human skin according to the frequency of application for a specific skin condition.

16. The method according to claim 15, wherein the skincare composition is applied to human skin on a daily basis.

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