Compositions and methods for improving the skin barrier

Abstract

The present invention relates to compositions comprising phytoene, phytofluene and zeta-carotene and methods of using same, such as for improving skin barrier function.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 211,072, entitled "Compositions and Methods for Improving Skin Barrier," filed June 16, 2021, the entire disclosure of which is incorporated herein by reference.

[0002] FIELD OF THEINVENTION The present invention relates generally to the field of carotenoids and methods of using same, such as to improve the skin barrier. [Background technology]

[0003] Tomato (Solanum lycopersicum) is a fruit consumed worldwide. Tomatoes are rich in nutrients such as vitamin C, potassium, essential amino acids and various antioxidants. Tomatoes are the primary dietary source of lycopene, a powerful antioxidant. Tomato consumption has been demonstrated to confer protection against oxidative stress-related diseases such as cancer and cardiovascular disease.

[0004] The use of dietary supplements is increasing worldwide. A survey of consumers in the United States from 2015 to 2019 revealed an upward trend in the use of various supplements. The most used supplements, in order of decreasing use, include fish oil, glucosamine / chondroitin, probiotics / prebiotics, melatonin, coenzyme Q10, echinacea, cranberry pills, garlic extract, ginseng, and ginkgo biloba. Some of these supplements have demonstrated health benefits, while others are controversial.

[0005] Lumenato™ is a dietary supplement prepared from the dried fruit of the yellow tomato. It is rich in carotenoids including lycopene (0.5 mg / capsule), 15-cis-phytoene (7.33 mg / capsule), phytofluene (2.06 mg / capsule), β-carotene (0.18 mg / capsule), γ-carotene (0.03 mg / capsule) and ζ-carotene (2.27 mg / capsule) as well as tocopherols (1.14 mg / capsule). Carotenoids and tocopherols in general are antioxidants that confer numerous health benefits.

[0006] Experiments conducted in the past 20 years to investigate the potential effects of oral carotenoid intake on the skin have focused primarily on the effects of β-carotene and lycopene, with less information on phytoene and phytofluene. Specific benefits of lycopene and / or β-carotene include protection from UV-induced erythema and sunburn, reduction of oxidative stress, and antioxidant / anti-inflammatory activity. β-carotene is naturally a precursor to vitamin A, thus supporting collagen synthesis. Lycopene has been found to prevent UV damage to cultured human skin fibroblasts. It has also been found to inhibit chemically induced skin cancer in animal models. It is believed that this ability, as well as its ability to ameliorate DNA damage caused by 4-hydroxyestradiol, may be due to its antioxidant properties. In addition to antioxidant activity, phytoene and phytofluene are colorless carotenoids that absorb ultraviolet light. This property may provide an additional level of protection to the skin.

[0007] Human skin is exposed to a variety of sources of oxidative stress, including ultraviolet radiation, ozone and other air pollutants, as well as resident microbial metabolism. These stressors accelerate skin aging, resulting in wrinkles, loss of elasticity and coarse texture.

[0008] Fibroblasts are the most abundant cells in the dermis. They synthesize collagen and maintain the extracellular matrix. During aging, collagen fibrils are gradually lost, resulting in wrinkles. Fibroblasts play an important role in innate immunity of the skin by producing cytokines to attract white blood cells to the dermis. Fibroblasts also communicate with melanocytes in the basal layer of the epidermis to affect melanin synthesis. Keratinocytes are the major cell type in the epidermis. They differentiate to produce the stratum corneum, which consists of flattened keratin-filled cells surrounded by a lipid matrix. The stratum corneum provides the permeability barrier of the skin. It functions to limit water and electrolyte loss and to restrict the entry of pollutants as well as harmful chemicals, bacteria and viruses into and through the skin. The lipids of the stratum corneum consist mainly of ceramides, cholesterol and long-chain fatty acids. A type of ceramide, the acylceramide, is rather unusual in that it contains a 30-34 carbon long ω-hydroxy acid amide-linked to a long chain base with a linoleic acid ester attached to the ω-hydroxyl group. These linoleic acid-containing lipids are essential for lipid organization and for barrier function.

[0009] There remains a significant need for compositions and methods for reducing transepidermal water loss (TEWL) and thus enhancing and / or improving the skin barrier. Summary of the Invention

[0010] The following embodiments and aspects thereof are described and illustrated in conjunction with systems, means and methods which are exemplary and illustrative and not limiting in scope.

[0011] The present invention is based, in part, in some embodiments, on the surprising discovery that ingestion of certain carotenoid compositions improved the skin barrier as demonstrated by a decrease in TEWL.

[0012] According to a first aspect, there is provided a method for treating a skin barrier associated disease or disorder in a subject in need thereof, comprising the step of administering to the subject a therapeutically effective amount of a composition comprising phytoene in an amount of 55-65% (w / w) of the total carotenoids in the composition, phytofluene in an amount of 10-20% (w / w) of the total carotenoids in the composition, and zeta-carotene in an amount of 15-25% (w / w) of the total carotenoids in the composition, thereby treating the skin barrier associated disease or disorder in the subject.

[0013] According to another aspect, there is provided a composition comprising phytoene in an amount of 55-65% (w / w) of the total carotenoids in the composition, phytofluene in an amount of 10-20% (w / w) of the total carotenoids in the composition, and zeta-carotene in an amount of 15-25% (w / w) of the total carotenoids in the composition, for use in treating a skin barrier associated disease or disorder in a subject in need thereof.

[0014] In some embodiments, the method further comprises a selecting step prior to the administering step, comprising determining a transepidermal water loss (TEWL) value of the subject's skin, wherein the TEWL value is at least 12 (g h / m 2 ) indicates that the subject is suitable for the treatment.

[0015] In some embodiments, TEWL values ​​are determined for the subject's facial skin, forehead skin, forearm skin, hand skin, palmar skin, leg skin, elbow skin, or any combination thereof.

[0016] In some embodiments, the skin barrier-associated disease is an epidermal skin disease or disorder.

[0017] In some embodiments, the skin barrier-associated disorder is characterized by an alteration in the lipid content, oxidative status, configuration of the stratum corneum, or any combination thereof.

[0018] In some embodiments, the skin barrier-associated disorder is characterized by a decrease in the amount of linoleic acid-containing acylceramides.

[0019] In some embodiments, the skin barrier associated disorder is selected from the group consisting of atopic dermatitis, psoriasis, Gaucher disease type II, Sjogren-Larsson syndrome, lamellar ichthyosis, X-linked ichthyosis, bullous ichthyosiform erythroderma, essential free fatty acid deficiency, acne vulgaris, age-related dry skin, anhidrotic ectodermal dysplasia, and atopic eczema.

[0020] In some embodiments, treating comprises reducing the TEWL, the number of pores, the area covered by pores, the level of redness, the area covered by redness, roughness, or any combination thereof, of the subject's skin.

[0021] In some embodiments, the decrease is at least a 5% decrease as compared to a control.

[0022] In some embodiments, the weight ratio of phytoene and phytofluene in combination:zeta-carotene ranges from 15:1 (w / w) to 2:1 (w / w).

[0023] In some embodiments, the composition further comprises an additional carotenoid selected from the group consisting of lycopene, β-carotene, γ-carotene, and any combination thereof.

[0024] In some embodiments, the composition comprises lycopene in an amount of less than 5% (w / w) of the total carotenoids in the composition, β-carotene in an amount of less than 5% (w / w) of the total carotenoids in the composition, γ-carotene in an amount of 0.2-1.5% (w / w) of the total carotenoids in the composition, or a combination thereof.

[0025] In some embodiments, the composition comprises a total carotenoid amount of 10-15% (w / w) of the composition.

[0026] In some embodiments, the composition further comprises tocopherol.

[0027] In some embodiments, the composition comprises tocopherol in an amount of 10-30% (w / w) of the total carotenoids in the composition.

[0028] In some embodiments, the composition further comprises a phytosterol.

[0029] In some embodiments, the composition comprises phytosterols in an amount of 5-15% (w / w) of the total carotenoids in the composition.

[0030] In some embodiments, administering includes orally administering, topically administering, or both.

[0031] Unless otherwise specified, all technical and / or scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, exemplary methods and / or materials are described below. In case of conflict, the present patent specification, including definitions, will control. In addition, the materials, methods, and examples are merely illustrative and are not necessarily intended to be limiting.

[0032] Further embodiments and the full scope of the present invention will become apparent from the detailed description given herein below. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

[0033] In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to the figures and by study of the following detailed descriptions. [Brief description of the drawings]

[0034] [Figure 1]1 includes a graph showing the effect of Golden Tomato Extract (GTE, also referred to herein as "Luminate™") on Transepidermal Water Loss (TEWL). A: High TEWL-GTE (N=18, 1 subject missed visit at week 8, p=0.428 at week 4, p=0.11 at week 8, p<0.001 at week 16), B: High TEWL-Placebo (N=14, 1 subject missed visit at week 8, p=0.382 at week 4, p=0.506 at week 8, p=0.293 at week 16), C: Low TEWL-GTE (N=14, p=0.129 at week 4, p=0.49 at week 8, p=0.421 at week 16), and D: Low TEWL-Placebo (N=15, 1 subject missed visit at weeks 8 and 16, p=0.099 at week 4, p=0.838 at week 8, p=0.231 at week 16). [Diagram 2] Includes "before" and "after" standard frontal view images of a panelist with a 53% improvement in TEWL. [Diagram 3] Figures 3A-C contain images of panelists "before" and "after" treatment showing a 26% reduction in TEWL. Figure 3A: front, Figure 3B: left, Figure 3C: right. Red areas are highlighted. [Figure 4] The left panel contains "before" and "after" images of a panelist with a 33% improvement in pore TEWL analysis. [Diagram 5] The right panel includes "before" and "after" treatment images of a panelist with a 48% improvement in pore TEWL analysis. [Figure 6] 6A-6B include left (FIG. 6A) and right (FIG. 6B) images of "before" and "after" treatment of a panelist with 21 and 33% improvement, respectively, in the TEWL analysis of texture. [Figure 7] The present disclosure includes non-limiting theoretical schemes of two mechanisms by which the golden tomato extracts disclosed herein enhance the skin's physical and biological barriers. [Figure 8]Figures 8A-8F include vertical bar graphs showing the mean scores of various parameters before and after 12 weeks of oral administration of Luminato capsules. As observed in Figure 8B, there was a highly significant (p<0.001) improvement in skin elasticity (21.54%) and firmness (27.18%) after 12 weeks of using the supplement. Figure 8A shows various measurements of skin tone. As observed in this graph, there was no improvement in pigmented spots, but there was still a 21% improvement in skin evenness (p<0.001). The skin appeared 29.75% lighter (p<0.001) and facial blemishes were reduced by 10.96% (p<0.001). Periocular dark circles also appeared to be reduced by 17.9% (p<0.001). FIG. 8C shows that after 12 weeks of supplement use, there was a 21.6% (p<0.001) perceived improvement in skin moisturization and an 18.18% (p<0.001) improvement in skin texture. FIG. 8D shows that after 12 weeks of supplement use, there was no improvement in deep wrinkles, but there was a significant (p<0.001) improvement of 12.97% in fine lines and wrinkles on the face. As observed in FIG. 8E, there was a slight improvement in the tendency of the skin to develop redness. The general appearance of aged / unhealthy skin was significantly (p<0.001) reduced by 23.53% after using the supplement. FIG. 8F shows the average score of overall skin condition before and after 12 weeks of treatment with the supplement. As observed in this graph, there was a highly significant (p<0.001) improvement of 28.22% in overall skin condition after using the supplement. [Figure 9]Figures 9A-9E include graphs showing results from the FACE-Q™ Satisfaction with Skin Scale before treatment, 4, 8 and 12 weeks after oral administration of Luminato capsules, and again 2 weeks after treatment was stopped. The data provided are from satisfaction scores, where a score of 1 was very dissatisfied, 2 was somewhat dissatisfied, 3 was somewhat satisfied, and 4 was very satisfied. The number of subjects reporting scores of 3 and 4 were counted and graphed as a percentage of the population showing a positive evaluation. Figure 9A shows "how my facial skin looks" first thing in the morning and at the end of the day. Initially, the scores were dominated by satisfied and dissatisfied, but after treatment there was a steady improvement from 84% to 75% by week 8 (p<0.001), after which it plateaued. The effect was maintained after 2 weeks of treatment was discontinued. Figure 9B shows a similar trend for the perception of a healthy and attractive appearance of the face. Steady improvements were observed for both of these parameters over the course of 12 weeks of treatment, at which point there was a statistically significant improvement of 74.6% (p<0.001) and 65.08% (p<0.001) for healthy and attractive skin, respectively. These perceptions continued to improve despite a 2-week cessation of treatment. Figure 9C: Perception of fresh and radiant face. Steady improvements were observed for these two parameters over the course of 12 weeks of treatment, at which point there was a statistically significant (p<0.001) change compared to baseline. These perceptions continued to improve despite a 2-week cessation of treatment. Figure 9D: Facial skin tone. Steady improvements were observed by week 8 in color evenness and tone from 61.9% to 74.6% (p<0.001) after treatment. By week 12 of treatment, over 71% (p<0.001) of the population declared the treatment to be beneficial for skin tone and evenness. After 2 weeks of treatment cessation, a further improvement of 81% and 72.41% was observed in skin tone evenness and tone, respectively. Figure 9E shows that 38.71% of the population noticed a reduction in skin redness after 4 and 8 weeks of supplement use, which increased to 41.94% after 12 weeks of supplement use.An improvement in the appearance and feel of healthy skin was observed by 35% of the population after 4 weeks of use, which increased to 56.45% and 58.06% after 8 and 12 weeks of use, respectively. The overall appearance of the skin improved in 35.48% of subjects after 4 weeks of use, which increased to 66.13% and 62.9% after 8 and 12 weeks, respectively. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0035] In some embodiments, the present invention relates to compositions comprising multiple carotenoids and methods of using the same. The present invention is based, in part, on the surprising discovery that tomato extracts containing high amounts of phytoene, phytofluene and zeta-carotene improve skin barrier and integrity in two complementary modalities: biological (innate immunity) and physical (stratum corneum barrier).

[0036] Treatment method According to some embodiments, there is provided a method for treating impaired skin barrier function in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition disclosed herein.

[0037] In some embodiments, the subject in need of treatment is a healthy subject suffering from a compromised barrier function, hi some embodiments, the subject in need of treatment is a healthy subject suffering from a compromised skin barrier.

[0038] In some embodiments, treating comprises improving the skin barrier or barrier function in the subject.

[0039] In some embodiments, treating comprises decreasing TEWL in the subject.

[0040] According to some embodiments, there is provided a method for treating a skin barrier associated disease or disorder in a subject in need thereof, comprising administering a therapeutically effective amount of a composition disclosed herein to the subject, thereby treating the skin barrier associated disease or disorder in the subject.

[0041] In some embodiments, the skin barrier disease or disorder comprises impaired barrier function or impaired skin barrier.

[0042] In some embodiments, the composition comprises phytoene in an amount of 55-65% (w / w) of the total carotenoids in the composition, phytofluene in an amount of 10-20% (w / w) of the total carotenoids in the composition, and zeta-carotene in an amount of 15-25% (w / w) of the total carotenoids in the composition.

[0043] In some embodiments, the method further comprises a selecting step that comprises determining a transepidermal water loss (TEWL) value of the subject's skin.

[0044] In some embodiments, the selecting step precedes the administering step.

[0045] In some embodiments, at least 1 (g h / m 2 ), at least 2(g h / m 2 ), at least 4(g·h / m 2 ), at least 5(g·h / m 2 ), at least 7(g·h / m 2 ), at least 10(g h / m 2 ), at least 11(g h / m 2 ), at least 12(g·h / m 2 ), at least 13(g·h / m 2 ), at least 14(g·h / m 2 ), at least 15(g·h / m 2 ), at least 16(g·h / m 2 ) or at least 20(g·h / m 2) or any value and range therebetween indicates that the subject is suitable for treatment according to the methods disclosed herein. Each possibility represents a separate embodiment of the present invention. In some embodiments, a TEWL value between 10 and 14 (g h / m 2 ), 1~15(g·h / m 2 ), 2~8(g·h / m 2 ), 3~12(g·h / m 2 ), 5~13(g·h / m 2 ), 4~10(g·h / m 2 ), 10~18(g·h / m 2 ), 11~20(g·h / m 2 ), 13~17(g·h / m 2 ) or 12~20(g·h / m 2 ) indicates that the subject is suitable for treatment according to the methods disclosed herein. Each possibility represents a separate embodiment of the present invention.

[0046] In some embodiments, subjects suitable for treatment according to the methods of the invention are characterized as having impaired and / or reduced barrier function and / or a reduced skin barrier.

[0047] In some embodiments, at least 10 (g h / m 2 ), 12(g·h / m 2 ), 14(g·h / m 2 ), 17(g·h / m 2 ) or 20(g h / m 2 TEWL values ​​of 12-20 (g·h / m 2 ) or any value and range therebetween are indicative of impaired and / or reduced barrier function and / or impaired skin barrier. Each possibility represents a separate embodiment of the present invention. In some embodiments, TEWL values ​​of 12-20 (g·h / m 2 ) or any value and range therebetween are indicative of impaired and / or reduced barrier function and / or impaired skin barrier. 2 ), 11~18(g·h / m 2 ), 12~19(g·h / m 2 ) or 10~20(g h / m 2 ) indicates impaired and / or reduced barrier function and / or reduced skin barrier. Each possibility represents a separate embodiment of the present invention.

[0048] In some embodiments, the TEWL value is determined for the subject's facial skin, forehead skin, forearm skin, volar forearm skin, hand skin, palmar skin, leg skin, elbow skin, or any combination thereof. In some embodiments, the TEWL value is determined for the subject's facial skin. In some embodiments, the facial skin comprises the subject's cheek.

[0049] As used herein, the term "transepidermal water loss" or "TEWL" refers to the loss of water from the body through the epidermis to the surrounding environment by diffusion and evaporation. The ability to measure TEWL is useful, for example, to detect damage to the skin indicative of a barrier disease or disorder.

[0050] Suitable methods and devices for determining skin TEWL are common and apparent to those skilled in the art and are discussed, for example, by Sotoodian and Maibach (2012) (Clinics in Dermatology).

[0051] In some embodiments, the subject is characterized by or afflicted by at least one of the following: nutritional deficiencies, reduced blood supply, exposure to radiation, oxidative stress, immunosuppression, psychological stress, dry skin, suffering from a chronic inflammatory disease, genetic predisposition, suffering from skin dysbiosis, reduced hygiene, excessive hygiene, reduced skin lipid content, sebum content or both, and any combination thereof.

[0052] In some embodiments, the radiation includes any wavelength of radiation within the optical spectrum. As used herein, the term "optical spectrum" refers to any wavelength within the optical spectrum. -9 m~10 -3 In some embodiments, the radiation wavelengths in the light spectrum include ultraviolet, visible, infrared, or a combination thereof. In some embodiments, the exposure to radiation includes exposure to sunlight.

[0053] As used herein, the term "ultraviolet (UV) radiation" encompasses any wavelength in the ultraviolet range. In some embodiments, UV is UV. In some embodiments, UV is UVA, UVB, UVC, or any combination thereof.

[0054] In some embodiments, the chronic inflammatory disease comprises a skin disease selected from acne, psoriasis, atopic dermatitis, contact dermatitis, and rosacea.

[0055] In some embodiments, the chronic inflammatory disease comprises a systemic inflammatory disease.

[0056] In some embodiments, the skin barrier-associated disease is a skin disease or disorder of the epidermis. In some embodiments, the skin barrier-associated disease is characterized by or includes an abnormality, deformation, pathological condition, or any combination thereof, of the epidermal layer of the skin. In some embodiments, the skin barrier-associated disease includes or includes an alteration of the stratum corneum of the epidermis. In some embodiments, the skin barrier-associated disease includes or includes an alteration or modification of the lipid content, oxidation state, configuration, or any combination thereof, of the stratum corneum.

[0057] In some embodiments, the change or alteration of the lipid content of the stratum corneum is such that it reduces the function of the stratum corneum, eg, impairs or reduces skin barrier activity.

[0058] In some embodiments, the skin barrier-associated disorder is characterized by a decrease in the amount of linoleic acid-containing acylceramides.

[0059] In some embodiments, the skin barrier associated disorder is selected from xerosis, atopic dermatitis, contact dermatitis, psoriasis, Gaucher disease type II, Sjogren-Larsson syndrome, lamellar ichthyosis, X-linked ichthyosis, bullous ichthyosiform erythroderma, essential free fatty acid deficiency, acne vulgaris, age-related dry skin, anhidrotic ectodermal dysplasia, atopic eczema, or any combination thereof.

[0060] As used herein, the term "skin barrier-associated disease or condition" encompasses any disease or condition in which a compromised skin barrier, a disrupted skin barrier, a non-functional skin barrier, a dysfunctional skin barrier, or any combination thereof, initiates, stimulates, propagates, increases, enhances, any equivalent thereof, or any combination thereof, an abnormal state in a subject, i.e., a deviation from homeostasis. In some embodiments, a skin barrier-associated disease includes any disease or condition that involves a compromised, disrupted, non-functional, dysfunctional, impaired or inhibited function of the skin barrier as part of the etiology and / or pathophysiology of the disease or condition.

[0061] In some embodiments, treating comprises reducing the TEWL, the number of pores, the area covered by pores, the level of redness, the area covered by redness, roughness, or any combination thereof, of the subject's skin.

[0062] In some embodiments, the decrease is at least 5%, at least 15%, at least 25%, at least 35%, at least 50%, at least 75%, at least 85%, at least 95%, at least 97%, at least 99%, or 100%, or any value and range therebetween. Each possibility represents a separate embodiment of the present invention. In some embodiments, the decrease is 5-50%, at least 1-25%, 10-55%, 20-95%, 50-99%, 40-75%, 5-85%, 30-95%, or 25-100% decrease. Each possibility represents a separate embodiment of the present invention.

[0063] The terms "subject" or "individual" or "animal" or "patient" or "mammal" as used herein, refers to any subject for which treatment is desired, particularly mammalian subjects, such as humans.

[0064] In some embodiments, the subject is at increased risk of developing acne. In some embodiments, the subject has an increased predisposition to developing acne. In some embodiments, the subject is characterized as having acne-prone skin.

[0065] In some embodiments, the decrease is compared to a control.

[0066] As used herein, the term "control" is interchangeable with "benchmark" or "baseline."

[0067] As used herein, a control comprises the skin of a healthy subject. In some embodiments, the control is a healthy skin sample derived from, isolated or obtained from the same subject (e.g., a subject afflicted with a skin barrier-associated disorder, including but not limited to, skin that comprises spots with limited areas of compromised barrier). In some embodiments, the control comprises healthy skin. In some embodiments, the healthy skin comprises the skin of a healthy subject. In some embodiments, the healthy skin comprises the skin of the same subject before the subject is afflicted with a skin barrier-associated disorder, as disclosed herein.

[0068] In some embodiments, administering comprises oral administration. In some embodiments, administering comprises topical administration. In some embodiments, administering comprises a combination of oral and topical administration.

[0069] As used herein, the phrase "inflammatory disease" refers to any disease that involves a multicomponent response of an organism (e.g., cells of the immune system, molecular signaling mediators such as cytokines, etc.) to a harmful foreign entity, such as a bacteria, fungus, virus, protozoa, allergen, pollutant, etc.

[0070] As used herein, the term "treatment" or "treating" a disease, disorder or condition includes alleviating at least one symptom thereof, reducing its severity or inhibiting its progression. Treatment does not necessarily mean completely curing the disease, disorder or condition. To be an effective treatment, a composition useful herein need only reduce the severity of the disease, disorder or condition, reduce the severity of symptoms associated therewith, or provide an improvement in the quality of life of the patient or subject.

[0071] As used herein, the term "prevention" of a disease, disorder or condition includes delaying, preventing, suppressing or inhibiting the onset of a disease, disorder or condition. The term "prevention" as used in accordance with the subject matter described herein relates to a prophylactic process in which a subject is exposed to a composition or formulation described herein prior to the induction or onset of the disease / disorder process. This can be done when an individual has a genetic pedigree that indicates a predisposition to the development of the disease / disorder to be prevented. For example, this may be the case for an individual whose ancestors indicate a predisposition to a particular type of, for example, inflammatory disease. The term "suppression" is used to describe a state in which the disease / disorder process has already begun but obvious symptoms of the condition have not yet been manifested. Thus, the cells of an individual may have the disease / disorder, but the apparent signs of the disease / disorder have not yet been clinically recognized. In either case, the term prevention can be applied to include both prevention and suppression. Conversely, the term "treatment" refers to the clinical application of an active agent to combat an existing condition whose clinical symptoms have already been manifested in the patient.

[0072] In some embodiments, preventing includes reducing the severity of the disease, delaying the onset of the disease, reducing the cumulative incidence of the disease, or any combination thereof.

[0073] According to some embodiments, a method is provided for modifying the expression level of a gene in a cell, comprising contacting the cell with a composition of the invention, thereby modifying expression of the gene.

[0074] In some embodiments, the gene is selected from IL6, CCL2, CXCL1, TNFPI3, DGAT2L6, ALOX5, BACH2, CYP4F11, CYP4A22, B3GALT1, B3GALT2, OR52N, OR5D15P, OR52B6, OR1F1, OR2L3, OR2V2, OR51AB1P, OR1B1, OR4C12, OR6C76, OR2AH1P, OR7A18P, OR1N2, OR10B1P, OR11L1, OR5BA1P, OR5M11, OR9A4, OR4U1P, or any combination thereof.

[0075] In some embodiments, the gene is selected from IL6, CCL2, CXCL1, TNFPI3, DGAT2L6, ALOX5, BACH2, CYP4F11, CYP4A22, B3GALT1, B3GALT2, or any combination thereof.

[0076] In some embodiments, the gene is selected from OR52N, OR5D15P, OR52B6, OR1F1, OR2L3, OR2V2, OR51AB1P, OR1B1, OR4C12, OR6C76, OR2AH1P, OR7A18P, OR1N2, OR10B1P, OR11L1, OR5BA1P, OR5M11, OR9A4, OR4U1P, or any combination thereof.

[0077] In some embodiments, modifying comprises increasing or decreasing.

[0078] In some embodiments, modifying comprises enhancing or decreasing.

[0079] In some embodiments, the methods comprise increasing the expression level of IL6, CCL2, CXCL1, TNFPI3, DGAT2L6, ALOX5, BACH2, CYP4F11, CYP4A22, B3GALT2, or any combination thereof.

[0080] In some embodiments, the methods comprise decreasing or reducing the expression level of B3GALT1.

[0081] In some embodiments, the method comprises increasing the expression level of IL6, CCL2, CXCL1, TNFPI3, DGAT2L6, ALOX5, BACH2, CYP4F11, CYP4A22, B3GALT2 or any combination thereof, and decreasing or reducing the expression level of B3GALT1.

[0082] In some embodiments, the method comprises increasing the expression level of OR52N, OR5D15P, OR52B6, OR1F1, OR2L3, OR2V2, OR51AB1P, OR1B1, OR4C12, OR6C76, OR2AH1P, OR7A18P, OR1N2, OR10B1P, OR11L1, or any combination thereof.

[0083] In some embodiments, the methods include decreasing or reducing the expression level of OR5BA1P, OR5M11, OR9A4, OR4U1P, or any combination thereof.

[0084] In some embodiments, the method includes increasing the expression level of OR52N, OR5D15P, OR52B6, OR1F1, OR2L3, OR2V2, OR51AB1P, OR1B1, OR4C12, OR6C76, OR2AH1P, OR7A18P, OR1N2, OR10B1P, OR11L1, or any combination thereof, and decreasing or reducing the expression level of OR5BA1P, OR5M11, OR9A4, OR4U1P, or any combination thereof.

[0085] In some embodiments, decreasing or reducing includes a decrease or reduction of at least 5%, at least 15%, at least 25%, at least 35%, at least 50%, at least 75%, at least 85%, at least 95%, at least 99% or 100%, or any value and range therebetween, with each possibility representing a separate embodiment of the present invention.

[0086] In some embodiments, enhancing or increasing includes an increase or enhancement of at least 5%, at least 25%, at least 50%, at least 75%, at least 100%, at least 150%, at least 250%, at least 500%, at least 750%, or at least 1,000%, or any value and range therebetween, with each possibility representing a separate embodiment of the present invention.

[0087] In some embodiments, the cell is a fibroblast. In some embodiments, the cell is a keratinocyte. In some embodiments, the cell is an epithelial cell. In some embodiments, the cell comprises a plurality of cells. In some embodiments, the plurality of cells comprises any combination of fibroblasts, keratinocytes, and epithelial cells. In some embodiments, the cell is a cell of a subject described herein.

[0088] composition According to some embodiments, there is provided a composition comprising phytoene in an amount of 55-65% (w / w) of the total carotenoids in the composition, phytofluene in an amount of 10-20% (w / w) of the total carotenoids in the composition, and zeta-carotene in an amount of 15-25% (w / w) of the total carotenoids in the composition for use in treating a subject suffering from a skin barrier associated disease.

[0089] In some embodiments, the composition comprises phytoene, phytofluene, and zeta-carotene. In one embodiment, the composition comprises phytoene, phytofluene, zeta-carotene, and additional carotenoids. In one embodiment, the composition comprises phytoene, phytofluene, zeta-carotene, lycopene, beta-carotene, gamma-carotene, tocopherol, and phytosterol.

[0090] In some embodiments, the carotenoid is a natural carotenoid extracted, isolated or purified from a fruit, vegetable or plant (including a plant part). In another embodiment, the carotenoid is a carotenoid extracted from a tomato plant. In another embodiment, the carotenoid is a carotenoid extracted from a tomato fruit. In another embodiment, the tomato carotenoid is a carotenoid-enriched tomato extract. In another embodiment, the tomato carotenoid is an all-natural carotenoid-rich tomato extract. In another embodiment, the tomato carotenoid is a tomato carotenoid complex. In another embodiment, the tomato carotenoid complex comprises a complex of phytonutrients including multiple carotenoids (such as phytoene, phytofluene, zeta-carotene, beta-carotene), tocopherols and phytosterols. In some embodiments, the carotenoid is a synthetic carotenoid.

[0091] In some embodiments, the present invention provides a tomato extract obtained by an innovative extraction protocol. This particular extract, which contains phytoene, phytofluene and zeta-carotene (in amounts specified below), has low cytotoxicity. In some embodiments, the low cytotoxicity is compared to other tomato extracts. In some embodiments, the low toxicity allows the composition of the present invention to be provided to a subject in need thereof in higher doses without causing a decrease in the subject's survival, well-being or both. In some embodiments, administering the composition of the present invention to a subject in need thereof allows the efficacy of treatment to be increased by providing active ingredients such as phytoene, phytofluene and zeta-carotene in higher amounts that enhance the therapeutic effect, without causing a decrease in the subject's survival, well-being or both due to high cytotoxicity.

[0092] In some embodiments, the compositions of the present invention provide higher amounts of carotenoids with lower toxicity compared to extracts from other plants, fruits or vegetables, such as tomatoes, In some embodiments, the compositions of the present invention provide enhanced therapeutic effects with lower toxicity compared to extracts from other plants, fruits or vegetables, such as tomatoes.

[0093] In some embodiments, the compositions of the present invention comprise natural carotenoids, synthetic carotenoids, or any combination thereof.

[0094] In some embodiments, the composition comprises phytoene in an amount of 10-40% (w / w), 15-35% (w / w), 20-45% (w / w), 25-35% (w / w), 20-30% (w / w), or 30-50% (w / w) of the total carotenoids of the composition, with each possibility representing a separate embodiment of the present invention.

[0095] In some embodiments, the composition comprises phytofluene in an amount of 1-10% (w / w), 3-12% (w / w), 4-14% (w / w), 5-10% (w / w), 8-15% (w / w), or 2-9% (w / w) of the total carotenoids of the composition, with each possibility representing a separate embodiment of the present invention.

[0096] In some embodiments, the composition comprises zeta-carotene in an amount of 4-20% (w / w), 6-18% (w / w), 5-15% (w / w), 6-12% (w / w), 9-17% (w / w), or 10-17% (w / w) of the total carotenoids in the composition, with each possibility representing a separate embodiment of the present invention.

[0097] In some embodiments, the weight ratio of phytoene and combination of phytofluene:zeta-carotene ranges from 20:1 (w / w) to 3:1 (w / w), 15:1 (w / w) to 3:1 (w / w), 20:1 (w / w) to 6:1 (w / w), 15:1 (w / w) to 2:1 (w / w), 17:1 (w / w) to 4:1 (w / w), 16:1 (w / w) to 7:1 (w / w), 13:1 (w / w) to 8:1 (w / w), 10:1 (w / w) to 3:1 (w / w), or 15:1 (w / w) to 10:1 (w / w), with each possibility representing a separate embodiment of the present invention.

[0098] In some embodiments, the composition further comprises an additional carotenoid. As used herein, "additional carotenoid" refers to any carotenoid or metabolite thereof other than, i.e., different from, phytoene, phytofluene, and zeta-carotene.

[0099] In some embodiments, the additional carotenoid is selected from lycopene, β-carotene, γ-carotene, and any combination thereof.

[0100] In some embodiments, the composition comprises lycopene in an amount of less than 10% (w / w), less than 7% (w / w), less than 5% (w / w), less than 3% (w / w), less than 2% (w / w), or less than 1% (w / w) of the total carotenoids in the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the present invention. In some embodiments, the composition comprises lycopene in an amount of 1-3% (w / w), 1-5% (w / w), 2-6% (w / w), 0.5-4.5% (w / w), 0.1-3% (w / w), 0.6-4.8% (w / w), or 2.5-4% (w / w) of the total carotenoids in the composition. Each possibility represents a separate embodiment of the present invention.

[0101] In some embodiments, the composition contains β-carotene in an amount of less than 10% (w / w), less than 7% (w / w), less than 5% (w / w), less than 3% (w / w), less than 2% (w / w), or less than 1% (w / w) of the total carotenoids in the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the present invention. In some embodiments, the composition contains β-carotene in an amount of 1-3% (w / w), 1-5% (w / w), 2-6% (w / w), 0.5-4.5% (w / w), 0.1-3% (w / w), 0.6-4.8% (w / w), or 2.5-4% (w / w) of the total carotenoids in the composition. Each possibility represents a separate embodiment of the present invention.

[0102] In some embodiments, the composition comprises γ-carotene in an amount of at least 0.15% (w / w), at least 0.18% (w / w), at least 0.2% (w / w), at least 0.25% (w / w), at least 0.35% (w / w), at least 0.5% (w / w), at least 0.75% (w / w), at least 0.9% (w / w), at least 1% (w / w), at least 1.2% (w / w), at least 1.35% (w / w), or at least 1.7% (w / w) of the total carotenoids in the composition, or any value and range therebetween. Each possibility represents a separate embodiment of the present invention. In some embodiments, the composition comprises γ-carotene in an amount of 0.15-3% (w / w), 0.2-2% (w / w), 0.2-1.5% (w / w), 0.5-3% (w / w), 0.7-1.6% (w / w), 0.4-2.8% (w / w), or 1.2-3.2% (w / w) of the total carotenoids in the composition, with each possibility representing a separate embodiment of the present invention.

[0103] In some embodiments, the composition comprises lycopene in an amount of less than 5% (w / w) of the total carotenoids in the composition, β-carotene in an amount of less than 5% (w / w) of the total carotenoids in the composition, γ-carotene in an amount of 0.2-1.5% (w / w) of the total carotenoids in the composition, or any combination thereof.

[0104] In some embodiments, the composition comprises additional carotenoid in an amount of 0.1-3% (w / w), 0.2-3.5% (w / w), 0.5-2.5% (w / w), 0.15-1.75% (w / w), 0.35-2.75% (w / w), 0.8-4% (w / w), 1-5% (w / w), or 1.5-4.75% (w / w), with each possibility representing a separate embodiment of the present invention.

[0105] In some embodiments, lycopene in an amount of less than 5% (w / w) of the total carotenoids in the composition.

[0106] In some embodiments, the composition comprises a total carotenoid amount of 5-25% (w / w), 10-15% (w / w), 12-35% (w / w), 3-17% (w / w), 2-20% (w / w), or 1-30% (w / w) of the composition, with each possibility representing a separate embodiment of the present invention.

[0107] In some embodiments, the composition further comprises a tocopherol (e.g., vitamin E). In some embodiments, the composition comprises tocopherol in an amount between 1-30% (w / w), 3-35% (w / w), 5-25% (w / w), 2-20% (w / w), 4-41% (w / w), 8-32% (w / w), or 13-39% (w / w) of the composition. Each possibility represents a separate embodiment of the present invention.

[0108] In some embodiments, the weight ratio of phytoene and combination of phytofluene:tocopherol ranges from 20:1 (w / w) to 3:1 (w / w), 15:1 (w / w) to 3:1 (w / w), 20:1 (w / w) to 6:1 (w / w), 17:1 (w / w) to 4:1 (w / w), 16:1 (w / w) to 7:1 (w / w), 13:1 (w / w) to 8:1 (w / w), 10:1 (w / w) to 3:1 (w / w), or 15:1 (w / w) to 10:1 (w / w), with each possibility representing a separate embodiment of the present invention.

[0109] In some embodiments, the weight ratio of zeta-carotene:tocopherol ranges from 3:1 (w / w) to 1:3 (w / w), 3:1 (w / w) to 1:2 (w / w), 3:1 (w / w) to 1:1 (w / w), 2:1 (w / w) to 1:1 (w / w), 2:1 (w / w) to 1:2 (w / w), 2:1 (w / w) to 1:3 (w / w), 1:1 (w / w) to 1:2 (w / w), or 1:1 (w / w) to 1:3 (w / w), with each possibility representing a separate embodiment of the present invention.

[0110] In some embodiments, the composition further comprises a phytosterol.

[0111] In some embodiments, the phytosterol is selected from cholesterol brassicasterol, campesterol, stigmasterol, β-sitosterol, δ5-avenasterol, δ7-avenasterol, δ7-stigmasterol, and any combination thereof.

[0112] In some embodiments, the composition comprises phytosterols in an amount of 1-20% (w / w), 2-19% (w / w), 10-25% (w / w), 5-25% (w / w), 8-16% (w / w), 6-18% (w / w), 3-20% (w / w), 4-17% (w / w), or 5-15% (w / w) of the composition. Each possibility represents a separate embodiment of the present invention. Each possibility represents a separate embodiment of the present invention.

[0113] In some embodiments, the weight ratio of phytoene and combination of phytofluene:phytosterol ranges from 20:1 (w / w) to 3:1 (w / w), 15:1 (w / w) to 3:1 (w / w), 20:1 (w / w) to 6:1 (w / w), 17:1 (w / w) to 4:1 (w / w), 16:1 (w / w) to 7:1 (w / w), 13:1 (w / w) to 8:1 (w / w), 10:1 (w / w) to 3:1 (w / w), or 15:1 (w / w) to 10:1 (w / w), with each possibility representing a separate embodiment of the present invention.

[0114] In some embodiments, the weight ratio of zeta-carotene:phytosterol ranges from 6:1 (w / w) to 2:1 (w / w), 5:1 (w / w) to 2:1 (w / w), 4:1 (w / w) to 2:1 (w / w), 3:1 (w / w) to 2:1 (w / w), 6:1 (w / w) to 3:1 (w / w), 5:1 (w / w) to 3:1 (w / w), 4:1 (w / w) to 3:2 (w / w), or 6:1 (w / w) to 3:1 (w / w), with each possibility representing a separate embodiment of the present invention.

[0115] Methods for determining the amount of phytonutrients such as carotenoids are common and will be apparent to one of skill in the art. Non-limiting examples of such methods include, but are not limited to, gas chromatography, liquid chromatography, and mass spectrometry.

[0116] In some embodiments, the composition is an oral composition or a topical composition. In some embodiments, the composition is a pharmaceutical or nutraceutical composition. In some embodiments, the composition comprises a pharmaceutical or nutraceutical acceptable carrier or excipient. In some embodiments, the composition is a cosmetic composition. In some embodiments, the composition comprises a cosmetic acceptable excipient. In some embodiments, the compositions described herein are orally ingestible dermatological compositions.

[0117] In some embodiments, the oral composition is in the form of a soft gel capsule. In some embodiments, the oral composition is in the form of a drink, injection, gummy, or powder. In some embodiments, the oral composition is mixed or blended into a foodstuff such as chocolate, ice cream, or other.

[0118] In some embodiments, the composition of the present invention is for use in improving skin barrier in a subject in need thereof.In some embodiments, the composition of the present invention is for use in treating or preventing skin barrier-related diseases in a subject in need thereof.In some embodiments, the composition of the present invention is for use in increasing the amount, concentration, or both of linoleic acid-containing acylceramide in the stratum corneum in a subject in need thereof.

[0119] In one embodiment, the compositions of the present invention can be provided to an individual per se. As used herein, the term "per se" refers to the composition not being further formulated and / or mixed with carriers, diluents, etc.

[0120] In one embodiment, the compositions of the present invention may be provided to an individual as part of a pharmaceutical or dietary supplement composition that includes a pharma- ceutically acceptable carrier.

[0121] In one embodiment, a "pharmaceutical composition," "cosmetic composition," or "nutraceutical composition" refers to the formulation of a composition described herein with other chemical ingredients, such as physiologically suitable carriers and excipients. The purpose of a pharmaceutical, cosmetic, or nutraceutical composition is to facilitate administration of the composition to an organism.

[0122] In some embodiments, a process is provided for producing a composition comprising phytoene in an amount of 20-30% (w / w), phytofluene in an amount of 5-10% (w / w), zeta-carotene in an amount of 5-15% (w / w), and an acceptable carrier. In some embodiments, the process comprises extracting golden tomatoes as disclosed herein. In some embodiments, the composition of the invention comprises a golden tomato extract produced by a process disclosed herein.

[0123] In one embodiment, the "combination preparation" specifically defines a "kit of parts" in the sense that it can be administered independently of the combination partners defined above or by using different fixed combinations with the identified amounts of the combination partners, i.e. simultaneously, in parallel, separately or sequentially. In some embodiments, some of the kit of parts can then be administered, for example, simultaneously or chronologically staggered, i.e. at different times, with equal or different time intervals for any part of the kit of parts. The total amount of the combination partners in the ratio can be administered in some embodiments in a combination preparation. In one embodiment, the combination preparation can be varied, for example, to address the needs of the treated patient subpopulation or the needs of one patient, which may be due to a specific disease, disease severity, age, sex or weight, as can be easily prepared by those skilled in the art.

[0124] In one embodiment, the phrases "physiologically acceptable carrier" and "pharmacologically acceptable carrier", used interchangeably, refer to a carrier or diluent that does not cause significant irritation to a mammal and does not abrogate the biological activity and properties of the composition being administered. Adjuvants are included in these phrases.

[0125] In one embodiment, "excipient" refers to an inert substance added to a composition to further facilitate administration of an active ingredient. In one embodiment, excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0126] Techniques for drug formulation and administration are described in the latest edition of Remington's Pharmaceutical Sciences (Mack Publishing, Easton, Pa.), the entire contents of which are incorporated herein by reference.

[0127] In one embodiment, suitable routes of administration include, for example, oral, rectal, mucosal, nasal, enteral delivery, or parenteral delivery, including intramuscular, subcutaneous and intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal or intraocular injections.

[0128] Unless otherwise indicated in this discussion, adjectives such as "substantially" and "about" modifying a condition or relationship characteristic of one or more features of an embodiment of the present invention are understood to mean defining that condition or characteristic within the tolerance permitted for the practice of the embodiment for its intended use. Unless otherwise indicated, the word "or" in this specification and claims is to be considered an inclusive "or" rather than an exclusive "or" and indicates at least one or any combination of the items it connects.

[0129] It will be understood that the terms "a" and "an" as used above and elsewhere in this specification refer to "one or more" of the listed components. It will be clear to one of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Thus, the terms "a," "an," and "at least one" are used interchangeably in this application.

[0130] To better understand the present teachings, and not to limit the scope of the present teachings in any way, unless otherwise indicated, all numbers and other numerical values ​​expressing quantities, percentages or ratios used in the specification and claims are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and appended claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should be construed in light of at least the number of reported significant digits and by applying ordinary rounding techniques.

[0131] In the specification and claims of this application, the verbs "comprise," "include," and "have," as well as their conjugations, are used to indicate that the object or objects of the verb are not necessarily an exhaustive recitation of the components, elements, or parts of the subject or subjects of the verb.

[0132] Other terms used herein are intended to be defined by their known meanings in the art.

[0133] Unless specifically stated otherwise or clear from the context, the term "or" as used herein is understood to be inclusive.

[0134] Throughout this specification and the claims, the word "comprise" or variations such as "comprises" or "comprising" indicate the inclusion of any recited integer or group of integers but not the exclusion of any other integer or group of integers.

[0135] As used throughout this specification and claims, the term "consists essentially of" or variations such as "consist essentially of" or "consisting essentially of" refers to the inclusion of any recited integer or group of integers, and refers to any inclusion of any recited integer or group of integers that does not materially alter the basic or novel characteristics of the method, structure, or composition specified.

[0136] As used herein, "comprises," "comprising," "containing," "having," and the like can mean "includes" and "including," and "consisting essentially of" or "consists essentially" likewise have the meanings given under U.S. patent law, which are open-ended and recognize the presence of unrecited items, but exclude prior art embodiments, so long as the basic or novel characteristics of the recited items are not altered by the presence of those items not recited. In one embodiment, the terms "comprises," "comprising," and "having" are interchangeable with "consisting."

[0137] Additional objects, advantages, and novel features of the present invention will become apparent to those skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, various embodiments and aspects of the present invention as described hereinabove and as claimed in the claims section below, find experimental support in the following examples. EXAMPLES

[0138] Generally, the nomenclature used herein and the laboratory procedures utilized in the present invention include chemical, molecular, biochemical and cell biological techniques, such techniques being fully explained in the literature. See, for example, Molecular Cloning: A laboratory Manual, Sambrook et al. (1989); Current Protocols in Molecular Biology, vols. I-III, edited by Ausubel, RM (1994); Cell Biology: A Laboratory Handbook, vols. I-III, edited by Cellis, JE (1994); The Organic Chemistry of Biological Pathways, by John McMurry and Tadhg Begley (Roberts and Company, 2005); Organic Chemistry of Enzyme-Catalyzed Reactions, by Richard Silverman (Academic Press, 2002); Organic Chemistry (6th ed.), by Leroy “Skip” G Wade; Organic Chemistry, TW See Graham Solomons and Craig Fryhle.

[0139] material and method Luminate™ (see Table 1 for a list of compounds) was provided by Lycored (Branchburg, NJ, USA). Tetrahydrofuran (THF) was purchased from Sigma Chemical (St. Louis, MO, USA). Primary human dermal fibroblasts obtained from the foreskin of one donor, DMEM, fetal bovine serum and XTT cell viability kit were purchased from Thermo Fisher Scientific (Branchburg, NJ, USA). Qiagen RNeasy kit for RNA extraction was purchased from Qiagen (Germantown, MD, USA). A 0.1% (w / v) stock solution of dried tomato extract in THF was prepared for use in gene expression experiments. Soft gel capsules containing Luminate™ (110 mg) were used for clinical trials. The control capsules for this study contained paraffin oil instead of tomato extract. [Table 1]

[0140] Gene expression Microarray analysis was used to evaluate the effect of two concentrations of Luminato™ extract: 0.001% and 0.0005% on gene expression in cultured human dermal fibroblasts.

[0141] Cytotoxicity assay Cytotoxicity was assessed using the XTT cell viability assay (Thermo Fisher Scientific) according to the manufacturer's instructions. Fibroblast cells in DMEM with 10% fetal bovine serum were seeded in 96-well plates and cultured overnight at 37°C under 5% CO2 and 95% relative humidity. The cells were incubated for 24 hours in the presence of 0.001% (v / v) and 0.0005% THF, THF + 0.001% (w / v) Luminate™, THF + 0.0005% Luminate™, and then incubated in these cell media for an additional 24 hours. Untreated cells were included as negative control. All treatments were performed in triplicate. An inhibition of viability of more than 20% of the control value was considered as cytotoxic. Test compounds were not toxic.

[0142] Treatment of human dermal fibroblasts Human dermal fibroblasts (HDFs) were cultured in 6-well plates (Thermo Fisher Scientific / Nunc) for 24 h and then treated with THF or Luminate™ for 6 h. After incubation, cells were washed with PBS and lysates were prepared for total RNA extraction using the Qiagen RNeasy kit according to the manufacturer's instructions. RNA concentration and purity were determined using a Nanodrop IMPLEN spectrophotometer (Westlake Village, CA, USA).

[0143] Total RNA quality control RNA isolated from epidermal tissue was shipped on dry ice to Advanced BioMedical Laboratories (Cinnaminson, NJ, USA) for processing. The overall integrity and RNA quality of the total RNA samples were confirmed by Advanced BioMedical Laboratories (Cinnaminson, NJ, USA). RNA integrity numbers (RIN) were calculated using a proprietary algorithm that takes into account several QC parameters (e.g., ribosomal RNA, 28S / 18S peak area ratio, unexpected peaks in the 5S region, etc.). An RIN number of 10 indicates perfect RNA quality, and an RIN number of 1 indicates degraded RNA. According to published data and our own experience, RNA with an RIN number greater than 8 is of sufficient quality for gene expression profiling experiments. All RNA samples had RIN numbers greater than 8. Microarray analysis was performed using Affymetrix Human Clariom D array processing.

[0144] Statistical analysis of gene expression data Differential gene expression was obtained using a threshold of 0.05 for statistical significance (p-value) and a LogFC (log fold change) of expression with an absolute value of at least 0.6. The fold increase or decrease of gene expression in treated cells versus control cells is asymmetric around zero. To avoid this problem, up- or down-regulation is usually expressed as the logarithm to the base 2 of the ratio. Thus, an absolute value of 0.6 represents a 1.5-fold up- or down-regulation. A value of 1 represents a 2-fold up- or down-regulation.

[0145] Human subjects One hundred and fourteen Japanese women aged 35 to 60 years were screened to find 66 eligible subjects. All eligible subjects had Fitzpatrick skin type 2 or 3 and Glogau skin classification type 2. Other inclusion criteria included the ability to understand the study and willingness to comply with the conditions of avoiding excessive UV exposure and not taking supplements that may interfere with the study. Of the 63 subjects who completed the study, two violated the protocol and their data were not analyzed. The study was conducted in accordance with the Declaration of Helsinki and was approved by the Aisei Hospital Ueno Clinical Research Ethics Committee of Japan (IRB#12000071). All subjects provided written informed consent before the start of the study. Subjects visited the clinic at baseline, week 4, week 8, and week 12 for photography, subjective assessment, and instrumental measurements. The clinical visit at week 12 was canceled due to the COVID-19 pandemic and was instead conducted at week 16. Subjects were to take one Luminart™ capsule per day with a meal.

[0146] Measurement of TEWL TEWL was measured under standard conditions at the time of the visit. A VapoMeter (Delfin Instruments, Tokyo, Japan) was used. Measurements were taken on the left cheek in the area from the corner of the eye and in the plane of the small protuberance of the external ear (tragus). For TEWL measurements, subjects were allowed to acclimate for 20 minutes in an environmental room at 21°C and 50% relative humidity, and then skin measurements were taken. All skin measurements were taken by the same person. Skin measurement locations were measured with a ruler to ensure the same location for each test point.

[0147] Photography and photo analysis Photographs of the subjects were obtained using the VISIA Evolution photography system. All VISIA modalities were used to obtain VISIA parameters, the primary outcome. Subject positioning was critical and had to be repeated at each time point. Features such as the subject's seat height and careful placement of the subject's chin and forehead within the imaging system were maintained. Subjects were also asked to remove hair from their faces, remove jewelry, and standardize clothing using black drapes. Frontal, left, and right views of the subject were obtained with eyes gently closed.

[0148] These images were further analyzed by Canfield Scientific (Canfield Scientific, New Jersey, USA) to extract and report numerical values ​​corresponding to various changes in the images, which are described below.

[0149] Red areas and texture A detailed analysis of the VISIA photographs was performed to detect red areas and textures by comparing the contrast of the area in question against its local zone. The acquired images were first normalized with respect to the background intensity distribution of the skin. Such a process eliminates illumination variations over the area of ​​interest. A contrast threshold is applied to select candidate areas that are darker than a minimum intensity level difference with respect to their surroundings. Finally, size and shape criteria are applied to screen the candidate areas and select the final spots.

[0150] Pore ​​diameter Pore ​​size was detected by detailed analysis of VISIA photographs. A pore analysis algorithm provides quantitative data and a scoring of visible facial pores. Pores were detected using an image analysis algorithm designed to detect circular objects within a specified diameter range that also meet a minimum circularity threshold as well as a minimum gradient threshold (comparing the intensity of the pore to its immediate surroundings). The analysis is performed on high-resolution images acquired by a Canfield research capture device (Canfield Scientific, New Jersey, USA).

[0151] Example 1: Gene Expression Analysis In this study, 97 genes were found to be significantly upregulated by treatment of human skin fibroblasts with 0.001% Luminate™ for 6 hours. For 55 of these genes, there is no readily available information on skin. Many of these were pseudogenes, and seven were olfactory receptor genes. 58 genes were significantly downregulated, and for 36 of these there was no readily available information on skin. 121 genes were upregulated and 44 genes were downregulated by 0.0005% Luminate™. There was some overlap with the findings for 0.001% Luminate™, but there were also some differences. Notably, 12 additional olfactory receptor genes were identified. The main genes to be considered are listed in Table 2, and the olfactory receptor genes are listed in Table 3. [Table 2] [Table 3]

[0152] Example 2: Clinical Trial - Effect of Luminate™ on TEWL In this clinical trial, 32 subjects had relatively high TEWL (12.5 g / h / m 2 16 subjects had TEWL values ​​above 16 weeks, and 29 subjects had lower TEWL values. Data from these two groups were analyzed separately. As shown in Figure 1, Luminate™ supplementation had no effect on low TEWL. However, in the high TEWL cohort, Luminate™ supplementation significantly reduced TEWL at 16 weeks.

[0153] Calculations of individual relative changes in TEWL for groups that showed significant changes ranged from 21-53% improvement from baseline. VISIA photographs of these panelists were further analyzed by Canfield Scientific for changes in skin appearance, i.e. redness, pores and texture. Selected photographs are provided herein.

[0154] These results represent selected photos from the supplement subgroup with TEWL baseline analyzed. The analysis reflects area ratio, which means the ratio of the total area occupied by the detected features to the area of ​​interest (area of ​​the mask). Feature detection is intensity / contrast based.

[0155] Figure 2 depicts standard view images of a panelist with a 35% improvement in TEWL values. When comparing the baseline photograph with the photograph obtained at week 16, the skin appears noticeably smoother and less visibly pore-like.

[0156] 3A-3C represent the red areas on the face of a panelist with a 26% improvement in TEWL values. When comparing the baseline photographs with those obtained for the skin at week 16, a clear reduction in redness is observed.

[0157] The red areas represent the presence of blood vessels and hemoglobin in the papillary dermis or lower layer of the skin, which gives these structures a red color, and are detected by RBX TECHNOLOGY (a trademark of Canfield Scientific), which has been shown to accurately detect and quantify skin erythema. With it, the red areas are photographed with cross-polarized light and outlined in light blue. The red dots represent the spots that are erythema and pimples, and the red color of the blood vessels is a linear feature. RBX TECHNOLOGY allows the device to see below the skin surface and visualizes certain conditions as related to both vascular disorders and hyperpigmentation. The red areas may be associated with various conditions such as inflammation or spider veins. Spider veins or broken capillaries are typically characterized as short thin lines that may be interconnected in a dense meshwork. Pimples or other conditions that produce inflammation vary in size but are generally smaller and round in shape. Large diffuse darker red areas may also indicate areas of concern. The core rationale of this technology is that hemoglobin occurs in vasculature in the papillary dermis, i.e., the lower layer of the skin, in oxidized and non-oxidized forms and contributes to the red coloring of skin tone.Some skin conditions, such as acne and rosacea, can cause organic changes in a person's vasculature and increase the level of hemoglobin present in the dermis.The increase in the amount of hemoglobin and the formation of new vasculature will cause red coloring and thus adversely affect the uniformity of skin tone.The results provided herein (Figure 3) show a clear reduction in the numerically calculated red area of ​​26% from baseline from the entire face.Interestingly, this panelist also showed a 26% reduction in TEWL value.

[0158] Pores are observed at the circular surface opening of the sweat duct. Each pore is outlined in dark blue, and shadowing makes the enlarged pores appear darker than the surrounding skin tone and are identified by their darker color and circular shape. The outlined areas identify the enlarged pores. Previously, the VISIA-CR method for pore assessment was demonstrated to have a high correlation with visual grading, resulting in pore size (r=0.86). It was also demonstrated to be suitable for discriminating between various age groups with different pore shapes and pore distributions.

[0159] Figure 4 represents the facial area of ​​a panelist with a 33% improvement in TEWL value. When comparing the baseline photograph of the skin with that obtained at week 16, a clear reduction in the number of pores is observed. This panelist also exhibited smoother skin.

[0160] Figure 5 represents the red areas on the face of a panelist with a 48% improvement in TEWL values. When comparing the baseline photograph of the skin with that obtained at week 16, a clear reduction in the number of pores is observed.

[0161] In addition to assessing pores, VISIA-CR photography can also allow for the assessment of changes in skin texture, whether it be coarser or smoother. For example, a study conducted to enhance skin penetration of Rhodobacter sphaeroides extract by sonophoresis resulted in an improvement in skin texture, among other attributes.

[0162] The texture analysis with VISIA is represented in a 2D photograph by the appearance of raised and depressed areas that indicate variations in the skin surface that affect the smoothness of the skin: raised areas are thus yellow and depressed areas are blue (Figure 6).

[0163] 6A-6B show the facial red areas of a panelist with a 21% improvement in TEWL values ​​and a panelist with a 33% improvement in TEWL values, respectively. When comparing baseline photographs of the skin with those obtained at week 16, a noticeable smoothing of the skin texture was observed.

[0164] Consideration Topical products have been used since ancient times to address various skin diseases and disorders. In recent decades, with the knowledge that the skin is nourished by blood, along with the belief that health equals beauty, the cosmetics and personal care industry has experienced a growth in dietary supplements that claim to improve the appearance of various skin characteristics, such as wrinkles, age spots, and redness. In summary, in vitro and in vivo studies to evaluate the effect of Luminate™ on the skin show that Luminate™ affects the skin barrier and integrity in two complementary modalities: biological (innate immunity) and physical (stratum corneum barrier). An illustration illustrating these two modalities is depicted diagrammatically in FIG. 7.

[0165] Many of the upregulated genes appear to enhance skin defense mechanisms, including enhancing innate immunity through recruitment of phagocytes and stimulating the production of antimicrobial fatty acids, enhancing DNA repair, and enhancing detoxification of xenobiotics.

[0166] IL-17 expression was not upregulated after 6 hours of Luminart™ treatment. Presumably, it was upregulated earlier, with upregulation of some of the genes in the IL-17 pathway occurring at 6 hours. These include IL-6, CCL2, and CXCL1. CCL2 and CXCL1 are chemokine genes that cause the attraction of neutrophils and monocytes. IL-17 and IL-6 act synergistically to increase the production of cell survival molecules under stress conditions. Similarly, TNF-α was not upregulated at 6 hours, but TNFIP3 (TNF-α-induced protein 3) was upregulated, indicating that the TNF genes were upregulated earlier. TNF-α stimulates collagen synthesis by dermal fibroblasts, which inhibits or possibly improves wrinkle formation. TNFIP3 is a deubiquitinase. It removes ubiquitin from cadherins to prevent their degradation. Taken together, these observations indicate that ingestion of Luminate™ activates both the IL-17 and TNF-α pathways in skin fibroblasts, leading to enhanced antimicrobial defense. These two pathways act synergistically in activating antimicrobial defense.

[0167] The gene product of DGAT2L6 increases sebum production, which may make lauric acid and sapienic acid more available at the skin surface, which are antibacterial substances.

[0168] Upregulated ALOX5 protein can act on arachidonic acid to generate 5-hydroperoxyeicosatetraenoic acid, which can then be converted to 5-hydroxyeicosatetraenoic acid (5-HETE) or leukotriene A4 (LTA4). LTA4 is quickly converted to leukotriene B4 (LTB4). 5-hydroxyeicosatetraenoic acid can be oxidized to 5-oxoeicosatetraenoic acid. LTB4, 5-HETE and 5-oxoeicosatetraenoic acid function as chemotactic factors to attract neutrophils and monocytes, thereby enhancing innate immunity. ALOX5 can also act on higher omega-3 essential fatty acids (eicosapentaenoic acid and docosahexaenoic acid), and some of these products may be anti-inflammatory.

[0169] BACH2 was significantly upregulated. This gene product is a marker of senescence and DNA damage. It has been implicated in B cell differentiation and immune system homeostasis.

[0170] CYP4F11 encodes a P450 enzyme that can protect against exogenous chemicals. This P450 has been found to hydroxylate many drugs. Interestingly, the expression of CYP4F11 was not significantly affected by treatment with 0.0005% Ruminate™ extract, but a different P450 gene, namely CYP4A22, was upregulated at this lower concentration. This enzyme has been found to ω-hydroxylate lauric and myristic acids, and to hydroxylate a variety of aliphatic and aromatic substrates.

[0171] A secondary but unexpected finding of this study was the upregulation of a set of genes encoding olfactory receptors. Olfactory receptors are G-protein coupled receptors. They have been found in a range of cell types other than olfactory neurons, including epidermal keratinocytes and melanocytes. In one study, an olfactory receptor (OR2AT4) was identified in keratinocytes that responded to a synthetic sandalwood odorant. Interaction of the odorant with the receptor resulted in calcium uptake. A later study investigated the role of this receptor in human hair shaft epithelial cells. In this case, activation of the receptor resulted in long-term hair growth ex vivo. Another study identified two other olfactory receptors in keratinocytes. OR2A4 / 7 responded to cyclohexyl salicylate, and OR51B5 responded to isononyl alcohol. Activation of either of these olfactory receptors resulted in an increase in intracellular calcium. Olfactory receptors were also detected in melanocytes. This receptor, OR51E2, responded to β-ionone, an odorous compound from violets, with an increase in intracellular calcium and stimulation of melanin synthesis. Although olfactory receptors have been reported in 3T3 cells, a cell line derived from embryonic fibroblasts, this study may be the first to identify a large number of olfactory receptor genes in human skin fibroblasts.

[0172] It was also notable that two galactosyltransferase genes were affected by Luminate™: one of these, B3GALT1, was significantly downregulated, while the other, B3GALT2, was upregulated.

[0173] Both of these β-1,3-galactosyltransferases transfer galactosyl residues from UDP-galactose to the terminal N-acetylglucosamine of glycoproteins or gangliosides. The B3GALT2 product can also transfer galactose to terminal galactose, but with a much lower affinity for this substrate. These changes in galactosyltransferase activity alter carbohydrate presentation at the cell surface. The consequences of this modification are currently unknown.

[0174] As described earlier, the lipids of the stratum corneum consist primarily of ceramides, cholesterol, and fatty acids. The fatty acids are primarily saturated species, 20-28 carbons long. Most of the ceramides also have a few easily oxidizable functional groups. These lipids are ideal for resisting oxidative damage. One exception is the linoleic acid-containing acylceramides, which are easily oxidized when exposed to air if not protected by antioxidants. Linoleic acid is essential for the barrier function of the skin. Linoleic acid from the circulation is absorbed by the epidermis and is first incorporated into a small pool of triglycerides. It is rapidly transferred to phosphoglycerides and then to acylglucosylceramides, which are the glycosylated precursors of linoleic acid-containing acylceramides. Glucose is β-glycosidically linked to the primary hydroxyl group of the long-chain base. The linoleic acid-containing acylglucosylceramides are associated with the lamellar granules. In the later stages of the keratinization process, the contents of the lamellar granules are extruded into the intercellular spaces, and glucosylceramides are deglycosylated to produce ceramides, including linoleic acid-containing acylceramides. Linoleic acid-containing acylceramides are essential for barrier function. Isolated linoleic acid-containing acylceramides are rapidly oxidized when exposed to air. Under optimal conditions, α-tocopherol is delivered to the skin surface via sebaceous gland secretions, which protect lipids and proteins from oxidation. Other antioxidants are also delivered to the sebum, but α-tocopherol is the predominant one in most environments. Under oxidative stress conditions, such as UV exposure, squalene, cholesterol, and keratin were found to be oxidized, and the oxidized keratin was found to be in the outer stratum corneum. In such impaired conditions, it seems likely that linoleic acid-containing acylceramides are also oxidized. This impairs the barrier function of the skin and causes an increase in TEWL. This may explain subjects with higher TEWL at baseline. Although some carotenoids diffuse from the dermis to the epidermis, the major portion of carotenoids in the dermis is delivered to the skin surface via sebum secretion and / or sweating, which may provide additional protection to acylceramides, thereby leading to restoration of barrier function with a reduction in TEWL.The results from Figure 1 show that after 16 weeks of Luminate™ supplementation, TEWL subjects have lower TEWL values ​​than placebo controls. If the linear trend continues, an additional 10 weeks of supplementation reduces TEWL to the same range as the low TEWL group. Error bars represent half the standard deviation with A and C increasing and B and D decreasing.

[0175] The lipids of the stratum corneum are organized into a multilamellar organization in the intercellular spaces of the stratum corneum with a major periodicity of 13 nm that is evident in transmission electron micrographs and X-ray diffraction data. This 13 nm periodicity is observed with reconstituted stratum corneum lipids, but in the absence of acylceramides, this periodicity is not seen by either electron micrographs or X-ray scattering. Mutations in enzymes specifically involved in acylceramide synthesis result in a variety of autosomal recessive ichthyoses with compromised barrier function. These genes include ELOV4, CYP4F22, PNPLA1, and ABHD5. The ELOV4 product is a fatty acid elongation enzyme that elongates C26-CoA to C30-CoA or greater. CYP4F22 encodes a P450 that ω-hydroxylates very long fatty acids. The PNPLA1 product transfers linoleic acid to the ω-hydroxyl group, and the ABHD5 product is a coactivator of PNPLA1. Also, in essential fatty acid deficiency, oleic acid replaces linoleic acid in acylceramides, and TEWL increases at least five-fold. In Gaucher disease, β-glucocerebrosidase, which deglycosylates acylglucosylceramides to produce acylceramides, is defective. In this situation, the ultrastructure of the intercellular spaces of the layer is altered, resulting in a marked increase in TEWL. These observations strengthen the notion that linoleic acid-containing acylceramides are essential for normal barrier function.

[0176] Astaxanthin is a carotenoid found in salmon, salmon roe and some shrimp, and has been reported to have benefits for skin health and homeostasis. In one randomized placebo-controlled study involving 36 male subjects, TEWL was significantly reduced in the astaxanthin group compared to the control after 6 weeks of daily intake of 6 mg of astaxanthin. Astaxanthin was also reported to result in improvements in mild wrinkles, age spot size, elasticity, moisturization and firmness. Another study found that astaxanthin increased the minimum erythema dose of UV after 9 weeks of astaxanthin supplementation at 4 mg / day. When TEWL was measured at the irradiated site, it increased in both the control and astaxanthin groups, but the increase was significantly smaller in the astaxanthin group.

[0177] Many studies have found that when TEWL is reduced by various interventions, it increases moisturization and smoothness of the skin surface, and reduces scaling and pore area. Similarly, in studies where skin is irritated by topical application of chemicals or use of harsh soaps, both TEWL and erythema are increased. Thus, TEWL correlates with many skin appearance parameters. In this study, a correlation was found between the reduction in TEWL and various attributes of skin appearance captured by VISIA photography and analyzed qualitatively and quantitatively. Specifically, it was demonstrated that the reduction in skin redness and skin pores, as well as an overall improvement in skin texture (reduced roughness) and appearance, were accompanied by a reduction in TEWL. Skin redness may be a sign of underlying inflammation at a subclinical level, a manifestation of a primary irritant, or an allergic reaction. Carotenoids are known anti-inflammatory and antioxidant agents, and thus the attenuation of skin redness could be explained by such activity, but the fact that the reduction in skin redness was strongly associated with a reduction in TEWL points to a potentially more complex multifunctional pathway. Moreover, the skin barrier is known to maintain a relatively robust homeostasis even when it is abnormal, meaning that it recovers to its baseline within hours to days in most cases when exposed to detergents, abnormal pH or other irritating conditions.

[0178] VISIA photographs show a reduction in concentrated darker red areas. These represent blood vessels and hemoglobin in the papillary dermis, the lower layer of the skin, giving these structures the red color that is detected by RBX TECHNOLOGY. The red areas may be associated with various conditions such as inflammation or spider veins. Spider veins or damaged capillaries are typically characterized as short thin lines that may be interconnected in a dense meshwork.

[0179] Application of topical retinoids to the skin has been shown to correlate with a decrease in TEWL and an improvement in skin texture. The similarity of effects may not be surprising, since carotenoids are oxidized precursors of retinoids.

[0180] This study evaluated the efficacy and safety of a new anti-aging cream containing retinal as a treatment for photographed skin. The study was conducted on 40 female Korean volunteers who applied the retinal cream twice daily for three months. Crow's feet wrinkles and skin texture were quantitatively evaluated using the Antera 3D® system. TEWL, skin moisturization, melanin index, and skin brightness were also evaluated. The results of this study demonstrated that the treatment improved the condition of photographed skin.

[0181] In summary, the research provided herein reveals the unique effect of Luminate™ on skin at the dermal and epidermal levels.These carotenoids and tocopherols strengthen the skin barrier and endow it with the means to deal with external stimuli.Furthermore, the inventor has demonstrated the correlation between barrier strength and certain skin appearance-related benefits, such as the reduction of redness and pores, as well as the effect of smoothing the skin.Furthermore, the inventor has demonstrated the presence of olfactory receptors in human skin fibroblasts and their significant upregulation by Luminate™.

[0182] Example 3: Effect of oral ingestion of Luminate on the improvement of visual and experimental skin attributes Healthy women aged 35-55 years who were interested in enhancing the health and appearance of normal facial skin were recruited for the study. The subjects downloaded the application (ClaimIt, Obvio Health USA), an online and mobile application interface for conducting clinical trials. This setup eliminated physical site visits and the study was conducted directly on each subject's mobile device. Participants signed an electronic informed consent (eIC) before beginning the screening process, after which they completed a study-specific screening questionnaire that included inclusion / exclusion criteria, including self-reported ethnic affiliation and skin type. Upon eligibility, they completed a medical history and concomitant medication form and participated in the study. The study product was shipped to eligible participants. Participants consumed one softgel daily before and after their main meal (breakfast, lunch or dinner) for 12 weeks. They were asked to complete the questionnaire within the application at pre-determined times during the study. A weekly e-diary was used to measure compliance with the study product intake as well as any reports of health or drug changes to determine adverse events, if any. At the end of the study (12 weeks of use), participants were instructed to safely discard any remaining softgels containing the study product and to complete a series of questionnaires regarding their experience with the study product. Two weeks after completing the treatment period, participants completed the FACE-Q™ Satisfaction with Skin (FACE-Q TM Participants completed the Study Satisfaction with Skin, End of Study Skin Questionnaire, and ClaimIt Experience Survey.

[0183] FIG. 8 shows the average scores of various parameters before and after 12 weeks of use of the supplement. FIG. 8A shows various measurements of skin tone. As observed in the graph, no improvement was observed in pigmented spots, but there was still an improvement in 21% of skin tone evenness (p<0.001). The skin was 29.75% lighter (p<0.001) and facial blemishes appeared to be reduced by 10.96% (p<0.001). Periocular dark circles also appeared to be reduced by 17.9% (p<0.001). FIG. 8 shows the average scores of various parameters before and after 12 weeks of use of the supplement. As observed in FIG. 8B, there was a highly significant (p<0.001) improvement in skin elasticity (21.54%) and firmness (27.18%) after 12 weeks of use of the supplement. FIG. 8A shows various measurements of skin tone. As can be seen in this graph, no improvement was observed in pigmented spots, but there was still a 21% improvement in skin evenness (p<0.001). The skin appeared to be 29.75% brighter (p<0.001) and facial blemishes were reduced by 10.96% (p<0.001). Periocular dark circles also appeared to be reduced by 17.9% (p<0.001). Figure 8C shows that after 12 weeks of supplement use, there was a 21.6% (p<0.001) perceived improvement in skin moisturization and an 18.18% (p<0.001) improvement in skin texture. Figure 4D shows that there was no improvement in deep wrinkles, but there was a significant (p<0.001) improvement of 12.97% in facial fine lines and wrinkles after 12 weeks of supplement use. As seen in Figure 8E, there was a slight improvement in the tendency of skin to develop redness. The general appearance of aged / unhealthy skin was significantly (p<0.001) reduced by 23.53% after using the supplement. Figure 8F shows the average score of overall skin condition before and after 12 weeks of treatment with the supplement. As seen in this graph, there was a highly significant (p<0.001) improvement of 28.22% in overall skin condition after using the supplement.

[0184] FIG. 5 shows results from the FACE-Q™ Satisfaction with Skin Scale before treatment, after 4, 8 and 12 weeks of treatment, and again after 2 weeks of treatment cessation. Data provided are from satisfaction scores, where a score of 1 was very dissatisfied, 2 was somewhat dissatisfied, 3 was somewhat satisfied, and 4 was very satisfied. The number of subjects reporting scores of 3 and 4 were counted and graphed as a percentage of the population showing a positive evaluation. FIG. 9A shows "how my facial skin looks" first thing in the morning and at the end of the day. Initially, the scores were dominated by satisfied and dissatisfied, but after treatment there was a steady improvement from 84% to 75% by week 8 (p<0.001), after which it plateaued. The effect was maintained after 2 weeks of treatment cessation. FIG. 9B shows a similar trend for the perception of a healthy and attractive appearance of the face. Steady improvements were observed for both of these parameters over the course of 12 weeks of treatment, at which point there was a statistically significant improvement of 74.6% (p<0.001) and 65.08% (p<0.001) for healthy skin and attractive skin, respectively. These perceptions continued to improve despite a 2-week cessation of treatment.

[0185] Figure 9C: Perception of healthy and radiant face. Steady improvements were observed for these two parameters over the course of 12 weeks of treatment, at which point changes were statistically significant (p<0.001) when compared to baseline. These perceptions continued to improve despite a 2-week hiatus from treatment.

[0186] FIG. 9D: Facial skin tone. Following treatment, there was a steady improvement in color evenness and tone by 8 weeks, ranging from 61.9% to 74.6% (p<0.001). By 12 weeks of treatment, over 71% of the population (p<0.001) declared the treatment to be effective for skin tone and shade evenness. After 2 weeks of treatment cessation, there was a further improvement of 81% and 72.41% in skin tone evenness and tone, respectively.

[0187] Figure 9E shows that 38.71% of the population noticed a reduction in skin redness after 4 and 8 weeks of supplement use, which increased to 41.94% after 12 weeks of supplement use. Improved appearance and feeling of healthy skin was observed by 35% of the population after 4 weeks of use, which increased to 56.45% and 58.06% after 8 and 12 weeks of use, respectively. Overall skin appearance improved in 35.48% of subjects after 4 weeks of use, which increased to 66.13% and 62.9% after 8 and 12 weeks, respectively.

[0188] Facial lines and wrinkles are the most visible signs of aging. It has been reported that furrows and wrinkles become progressively deeper in the skin of individuals with low antioxidant levels. In the current study, a significant reduction in fine lines and wrinkles was observed after 12 weeks of product use. The majority of subjects in the study showed very low scores for deep wrinkles, yet 61% of the subject population noticed an improvement in wrinkles after treatment with the supplement.

[0189] Example 4: Clinical Trial to Evaluate the Efficacy and Consumer Perception of Oral Supplements The primary objective of this clinical trial is to evaluate the efficacy of an oral supplement to alter facial skin characteristics after 4, 8 and 12 weeks of use. The secondary objective is to obtain consumer perception of the test product through the use of a questionnaire.

[0190] Approximately 66 subjects participated in this clinical trial to evaluate the effectiveness of oral supplements to change facial skin characteristics after 4 and 12 weeks of use and to obtain consumer perception of the test product through the use of questionnaires. Study assessments included a comprehensive metabolic panel, Cutometer measurements, Canfield's VISIA-CR images, VapoMeter measurements, and a consumer perception questionnaire.

[0191] VapoMeter The VapoMeter is the only fully portable device available for the measurement of TEWL (Transepidermal Water Loss) values ​​and evaporation rates. TEWL is a well-known index of skin barrier function. Two VapoMeter measurements are obtained from randomized cheeks at each designated time point.

[0192] Cute Meter The Cutometer is a non-invasive method for assessing the mechanical properties of skin. The instrument measures the vertical deformation of the surface of the skin when tensioned by vacuum suction (500 mmHg) through a small probe opening.

[0193] The following parameters can be measured: final elongation Uf (R0) measured in 10 seconds, immediate elongation Ue measured in 0.1 seconds, delayed elongation Uv, immediate contraction Ur. The deformation parameters are extrinsic parameters that depend on the thickness of the skin. To avoid measuring the thickness of the skin, the following ratio is used to evaluate the elasticity of the skin: R0: This parameter describes the passive behavior of the skin against forces. A decrease in this parameter indicates an improvement in firmness. R1: Ability of the skin to return to its original state. A decrease in this parameter indicates an improvement. R2: Total elasticity. The closer this value is to 1 (100%), the more elastic it is. This curve is a very important parameter. R2 is the elasticity of the skin and its ability to return to its original position. An increase in this parameter indicates an improvement in elasticity. R3: The final curve compared to the maximum amplitude of the first curve. The skin fatigue effect is visible as its amplitude increases with each new suction. R3 correlates with skin fatigue. Repeated suction and release by the cutometer causes skin fatigue, resulting in a decrease in elasticity and an increase in maintaining the deformed position. A decrease in this parameter indicates an improvement. R4: Final minimum amplitude compared to the first curve. A skin fatigue effect is visible as the ability to reshape decreases with each new suction. R4 correlates with skin fatigue. Repeated suction and release causes skin fatigue according to the Cutometer, resulting in a decrease in elasticity and an increase in maintaining the deformed position. A decrease in this parameter indicates an improvement. Ur / Ue (R5) is the biological elasticity of the skin. It measures the ability of the skin to regain its initial configuration after deformation. A value of 1 indicates 100% elasticity. An increase in this ratio indicates improved elasticity.

[0194] Uv / Ue (R6) is the ratio between delayed and immediate deformation, i.e. between viscoelastic and elastic. An increase in the value of this ratio indicates that there has been an increase in the viscoelastic part of the deformation and / or a relative decrease in the elastic part. A decrease in these ratios indicates an improvement in elasticity.

[0195] Ur / Uf (R7) is a measure of the net elasticity of the skin. An increase in this ratio indicates improved elasticity.

[0196] Cutometer measurements are taken for R1, R2, R3, R4, R5, R6, R7 and R0.

[0197] Measurements are taken by holding a probe firmly against the surface of the skin, a vacuum pump sucks the skin into a small opening and the probe measures the height to which the skin stretches.

[0198] Three measurements are generated automatically by performing three 20 second cycles. Each cycle consists of a suction phase where the skin is sucked into the probe, and a relaxation phase where the suction is stopped and the skin is released. The data is electronically imported into a computer where ratio values ​​are calculated. The readings are displayed and then manually transcribed onto a score sheet.

[0199] One Cutometer measurement is obtained from the randomized cheek at each designated time point.

[0200] CANFIELD'S VISIA-CR Canfield's VISIA-CR's sophisticated technology produces high-quality, reproducible facial images that can demonstrate treatment efficacy through subjective clinical assessment of skin features such as wrinkles and fine lines, skin texture, coloration / evenness, photodamage, vascular features and porphyrins (Propionibacterium acnes).

[0201] VISIA-CR imaging will be acquired with eyes closed. Subjects will be asked to wear a headband to keep hair from falling forward and a cape to cover clothing. Subjects may be asked to remove jewelry at the technician's instruction.

[0202] Assessment of the following parameters is obtained: fine lines and wrinkles (crow's feet, forehead, cheeks), pores, age spots, pigmentation / evenness, texture / roughness and porphyrins.

[0203] The study will involve 66 subjects at three time points, and analysis will be performed on VISIA images of all subjects.

[0204] Details of the study are as follows: 66 subjects: 33 in the active group and 33 in the placebo group Two products (active vs. placebo) Three time points: baseline, week 4 and week 12 Acquisition of left, front and right images Crossed, parallel, UV and standard polarized (all Visia CR modalities)

[0205] The measurement and lighting systems must be the same throughout the study, and subjects must be very well repositioned between time points.

[0206] Analysis of the outer corner of the eye To assess the effectiveness of the product on the crow's feet, the analysis is performed on left and right images.

[0207] Definition of regions of interest in each canthodontic area (left and right).

[0208] Spatial registration between images of each subject to track precisely the same region over time.

[0209] Research and optimization of extraction parameters for crow's feet wrinkles on several images.

[0210] Extraction of crow's feet wrinkles with optimized parameters on all images.

[0211] Extraction of crow's feet wrinkle parameters: visible surface, visible depth, visible volume and visible length.

[0212] Forehead Wrinkle Analysis To evaluate the effectiveness of the product against forehead wrinkles, the analysis is performed on facial images.

[0213] Defining the area of ​​interest on the forehead

[0214] Spatial registration of regions between images of each subject to accurately track the same regions over time

[0215] Study and optimization of extraction parameters for forehead wrinkles on several images

[0216] Extraction of forehead wrinkles with optimized parameters on all images

[0217] Extraction of forehead wrinkle parameters: visible surface, visible depth, visible volume and visible length.

[0218] Analysis of fine lines on cheeks To assess the effectiveness of the product on the cheeks, the analysis is performed on the left and right images.

[0219] Definition of regions of interest on each cheek (left and right).

[0220] Spatial registration of regions between images of each subject to accurately trace the same regions over time.

[0221] Research and optimization of extraction parameters for cheek fine lines on several images.

[0222] Extraction of cheek fine lines with optimized parameters on all images.

[0223] Extraction of cheek fine lines parameters: visible surface, visible depth, visible volume and visible length.

[0224] Color and uniformity analysis To assess the effectiveness of the product on skin tone and blemishes, the analysis is performed on frontal, left and right images.

[0225] Selection of one area on the cheek (right and left profile) and one area on the forehead (frontal) on each photograph at baseline.

[0226] Spatial registration of regions between images of each subject to accurately trace the same regions over time.

[0227] Data extraction of colorimetric parameters: L * , a * , b * , ITA° and IWA° Newtone (whitening index).

[0228] Skin Texture Analysis To assess the effectiveness of the product on skin roughness, the analysis is performed on frontal, left and right images.

[0229] Selection of one area on the cheek (right and left profile) and one area on the forehead (frontal) on each photograph at baseline.

[0230] Spatial registration of regions between images of each subject to accurately trace the same regions over time.

[0231] Haralick extraction parameters: contrast and entropy for skin smoothness and texture regularity.

[0232] Calculation of 2D roughness parameters: Spa, Spq and Sdev.

[0233] Pore ​​Analysis To assess pores, the analysis is performed on the en face image.

[0234] Definition of regions of interest near the nose on the left and right sides of the face.

[0235] Spatial registration of regions between images of each subject to accurately trace the same regions over time.

[0236] Study and optimization of extraction parameters for pores on several images.

[0237] Pore ​​extraction with optimized parameters on all images.

[0238] Characterization of the extracted areas: number, visible surface, visible depth and visible volume analysis.

[0239] Porphyrin Analysis To assess porphyrins, the analysis is performed on the upper cheek (near the nose) on a facial image.

[0240] Definition of one region of interest at and near the nose.

[0241] Spatial registration of regions to analyze the same area over time.

[0242] Parameter tuning for porphyrin detection and segmentation.

[0243] Extraction of porphyrins with optimized parameters on all images.

[0244] Quality control for all segmented images.

[0245] Analysis of porphyrins: number, amount and fluorescent surface of elements.

[0246] Analysis of spots To evaluate the effectiveness of the product on dark spots, the analysis is performed on the left and right images.

[0247] Selection of overall pigmented spots in each subject on the two cheeks at T0.

[0248] Detection and automatic segmentation of pigment spots at all time points.

[0249] Surface extraction of pigment spots.

[0250] Pigment spot color (L * , a * , b * , ITA°, IWA°Newtone) extraction, contrast between surrounding skin and pigment spots and surrounding skin.

[0251] Consumer Perception Questionnaire Evaluation of test material attributes and efficacy of the test material can be determined by questioning treated subjects regarding consumer perception after use and efficacy of the test material. Administration of questionnaires using EvaSys or similar allows an efficient and accurate method for determining subject response rates and determining consensus of the clinical trial population.

[0252] Comprehensive Metabolic Panel Blood is drawn from each subject for a comprehensive metabolic panel that includes the following parameters: glucose, calcium, protein (albumin and total protein), electrolytes (sodium, potassium, carbon dioxide and chloride), kidney tests (blood urea nitrogen and creatinine), and liver tests (alkaline phosphatase, alanine aminotransferase (also known as SGPT), aspartate aminotransferase (also known as SGOT), and bilirubin).

[0253] Phlebotomy procedures will be performed according to CRL SOP AC1.3. A comprehensive metabolic panel will provide liver and renal function results to ensure safety of test material for the duration of the study. Subjects with values ​​outside acceptable ranges will be withdrawn or discontinued.

[0254] Blood draws will be performed by a licensed technician at baseline and at week 4. Samples will be sent to the laboratory for metabolic panel.

[0255] D-squame tape stripping method The shedding or accumulation of stratum corneum in flakes is called desquamation. Under normal conditions, the epidermis is completely replaced every 25-30 days depending on the site of the skin. Corneum cells pile up and become the stratum corneum at the end of the keratinization process. Abnormal shedding caused by underlying conditions like parakeratosis (psoriasis) or loss of lipids and moisture (xeroderma) leads to desquamating skin eruptions. The D-Squame is a 22mm diameter crystal clear adhesive coated disk shaped sampling device. It has a homogenous layer of medical grade adhesive that safely removes the surface keratinocytes and provides optimal visibility for adhering skin cells.

[0256] Tape Stripping Method A series of tape strips are obtained successively from the same site to remove several layers of the stratum corneum. Tape stripping is performed using a D-Squame standard sampling disk (a circular 22 mm adhesive disk).

[0257] Study Procedure: Tape stripping is performed on the randomized right or left cheek. A D-Squame standard sampling disk is applied to the randomized cheek and then removed three times in succession, a new tape is used each time the method is used, and TEWL readings are taken after each of the three tape strips. This is done using a 20 g / m 2 Repeat until a TEWL reading of 100 / hr is reached.

[0258] Baseline Subjects will present to the testing facility with a clean face and no make-up. A photo release form will be completed by the subject. Informed consent will be obtained. Inclusion / exclusion criteria will be confirmed. Subjects will be allowed approximately 15 minutes (± 5 minutes) to acclimate to ambient laboratory conditions.

[0259] The following laboratory evaluations will be performed: comprehensive metabolic panel, Cutometer measurements, Canfield VISIA-CR imaging, and VapoMeter measurements.

[0260] ECRL Technician 20g / m 2 A tape strip of the left or right randomized cheek will be performed using a D-squame until a TEWL reading of 100 / hr is reached. Subjects will be instructed to return to the testing room approximately 3 hours after tape stripping. Subjects will be instructed to refrain from washing or applying any products to their face.

[0261] 3 hours (± 30 minutes) after tape stripping Subject is allowed approximately 15 minutes (± 5 minutes) to acclimate to ambient laboratory conditions. ECRL technicians will obtain VapoMeter readings.

[0262] Week 4 Subjects will return to the testing facility with a clean face and no makeup. Subjects will be allowed approximately 15 minutes (± 5 minutes) to acclimate to the ambient laboratory conditions. The following study assessments will be administered: Comprehensive Metabolic Panel, Cutometer measurements, Canfield VISIA-CR Imaging, VapoMeter measurements, and Consumer Perception Questionnaire.

[0263] ECRL Technician 20g / m 2 A tape strip of the left or right randomized cheek will be performed using a D-squame until a TEWL reading of 100 / hr is reached. Subjects will be instructed to return to the testing room approximately 3 hours after tape stripping. Subjects will be instructed to refrain from washing or applying any products to their face.

[0264] 3 hours (± 30 minutes) after tape stripping Subject is allowed approximately 15 minutes (± 5 minutes) to acclimate to ambient laboratory conditions. ECRL technicians will obtain VapoMeter readings.

[0265] Week 8 Subject returns to testing facility.

[0266] The following research assessments will be carried out: Consumer Perception Questionnaire.

[0267] Week 12 (last visit) Subjects will return to the testing facility with a clean face and no makeup. Subjects will be allowed approximately 15 minutes (± 5 minutes) to acclimate to ambient laboratory conditions. Daily diaries will be reviewed and collected for study compliance.

[0268] The following study evaluations will be performed: Cutometer measurements, Canfield's VISIA-CR images, VapoMeter measurements and a consumer perception questionnaire.

[0269] ECRL Technician 20g / m 2 A tape strip of the left or right randomized cheek will be performed using a D-squame until a TEWL reading of 100 / hr is reached. Subjects will be instructed to return to the testing room approximately 3 hours after tape stripping. Subjects will be instructed to refrain from washing or applying any products to their face.

[0270] 3 hours (± 30 minutes) after tape stripping Subject is allowed approximately 15 minutes (± 5 minutes) to acclimate to ambient laboratory conditions. ECRL technicians will obtain VapoMeter readings.

[0271] Although the invention has been described in detail, those skilled in the art will appreciate that many variations and modifications may be made. Accordingly, the invention should not be construed as being limited to the particularly described embodiments, but rather the scope and spirit of the invention will be more readily understood by reference to the following claims.

Claims

**Claim 1**: A composition for use in the treatment of skin barrier-related diseases or disorders in a subject in need thereof, the composition comprising phytoene in an amount of 55-65% (w / w) of the total carotenoids in the composition, phytofluene in an amount of 10-20% (w / w) of the total carotenoids in the composition, and ζ-carotene in an amount of 15-25% (w / w) of the total carotenoids in the composition. **Claim 2**: The subject is selected for said treatment prior to administration based on determination of the trans-epidermal water loss (TEWL) value of the skin of the subject, where a TEWL value of at least 12 (g·h / m ２ ) indicates that the subject is suitable for said treatment, the composition for use according to claim 1. **Claim 3** The TEWL value is determined for the skin of the face, forehead, forearm, hand, palm, leg, elbow of the subject or any combination thereof, the composition for use according to claim 2. **Claim 4** The skin barrier-related disease is an epidermal skin disease or disorder, the composition for use according to claim 1. **Claim 5** The skin barrier-related disease is characterized by a change in the lipid content, oxidation state, arrangement of the stratum corneum or any combination thereof, the composition for use according to claim 1. **Claim 6** The skin barrier-related disease is characterized by a decrease in the amount of linoleic acid-containing acyl ceramides, the composition for use according to claim 1. **Claim 7** The skin barrier-related disease is selected from the group consisting of atopic dermatitis, psoriasis, Gaucher disease type II, Sjogren-Larsson syndrome, lamellar ichthyosis, X-linked ichthyosis, bullous ichthyosiform erythroderma, essential free fatty acid deficiency, acne vulgaris, senile dry skin, anhidrotic ectodermal dysplasia and atopic eczema, the composition for use according to claim 1. **Claim 8** The combination of the phytoene and the phytofluene: the weight ratio of the ζ-carotene is in the range of 15:1 (w / w) to 2:1 (w / w), the composition for use according to claim 1.

9. The composition further comprises a further carotenoid selected from the group consisting of lycopene, β-carotene, γ-carotene and any combination thereof, the composition for use according to claim 1.

10. The composition comprises the lycopene in an amount of less than 5% (w / w) of the total carotenoids in the composition, the β-carotene in an amount of less than 5% (w / w) of the total carotenoids in the composition, the γ-carotene in an amount of 0.2 to 1.5% (w / w) of the total carotenoids in the composition or a combination thereof, the composition for use according to claim 9.

11. The composition comprises a total carotenoid amount of 10 to 15% (w / w) of the composition, the composition for use according to claim 1.

12. The composition further comprises tocopherol, the composition for use according to claim 1.

13. The composition comprises the tocopherol in an amount of 10 to 30% (w / w) of the total carotenoids in the composition, the composition for use according to claim 12.

14. The composition further comprises phytosterol, the composition for use according to claim 1.

15. The composition comprises the phytosterol in an amount of 5 to 15% (w / w) of the total carotenoids in the composition, the composition for use according to claim 14.

16. The treatment comprises oral administration, topical administration or both of the composition, the composition for use according to claim 1.

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