阅读说明：本技术 用于处理玫瑰痤疮和皮肤发红的局部皮肤用组合物 (Topical skin composition for treating rosacea and redness of the skin ) 是由 蒂凡尼·卡尔 大卫·甘 吉赛·卡拉哈斯蒂 于 2020-02-17 设计创作，主要内容包括：本发明一般涉及用于处理皮肤的使用方法和组合物。组合物包含海枣(Phoenix dactylifera)提取物、茶树精油、密罗木(Myrothamnus flabellifolia)提取物和糖类同分异构体的组合。该组合可用于制造局部皮肤用组合物,所述局部用组合物减少玫瑰痤疮、红斑和/或炎症,并抑制一氧化氮合酶、增加α2A肾上腺素能受体激动剂活性、减少氧化、增加组合物或皮肤的抗氧化能力、抑制环氧合酶-2(COX-2)的产生、抑制血管内皮生长因子(VEGF)的产生、抑制白细胞介素-6(IL-6)和白细胞介素-8(IL-8)的产生、减少肿瘤坏死因子α(TNF-α)的产生、增加胶原蛋白刺激、增加赖氨酰氧化酶的表达、抑制基质金属蛋白酶1(MMP1)活性、增加闭合蛋白的产生、增加丝聚蛋白的产生和增加皮肤保湿。该组合还可以用于减少暂时性或持续性红斑、毛细管扩张、炎性丘疹和/或脓疱、暂时性或持续性皮肤潮红和/或缔结组织的增生。(The present invention relates generally to methods of use and compositions for treating skin. The composition comprises a combination of date (Phoenix dactylifera) extract, tea tree essential oil, Artocarpus heterophyllus (Myrothamnus flabellifolia) extract and saccharide isomerate. The combination can be used to make a topical skin composition that reduces rosacea, erythema and/or inflammation and inhibits nitric oxide synthase, increases alpha 2A adrenergic receptor agonist activity, reduces oxidation, increases the antioxidant capacity of the composition or the skin, inhibits cyclooxygenase-2 (COX-2) production, inhibits Vascular Endothelial Growth Factor (VEGF) production, inhibits interleukin-6 (IL-6) and interleukin-8 (IL-8) production, reduces tumor necrosis factor alpha (TNF-alpha) production, increases collagen stimulation, increases lysyl oxidase expression, inhibits matrix metalloproteinase 1(MMP1) activity, increases occludin production, increases filaggrin production, and increases skin moisturization. The combination may also be used to reduce temporary or persistent erythema, telangiectasia, inflammatory papules and/or pustules, temporary or persistent skin flushing, and/or proliferation of connective tissue.)

1. A method of treating skin comprising topically applying to the skin an effective amount of a topical composition comprising date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate, wherein the skin is treated.

2. The method of claim 1, wherein the skin is treated to reduce rosacea, erythema and/or inflammation, and wherein the rosacea, erythema and/or inflammation is reduced.

3. The method of claim 1, wherein the skin is treated to reduce transient or persistent erythema, telangiectasia, inflammatory papules and/or pustules, transient or persistent skin flushing, and/or hyperplasia of connective tissue, and wherein the transient or persistent erythema, telangiectasia, inflammatory papules and/or pustules, transient or persistent skin flushing, and/or hyperplasia of connective tissue is reduced.

4. The method of claim 1, wherein the skin is treated to inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of the skin, inhibit cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) production, decrease tumor necrosis factor alpha (TNF-alpha) production, increase collagen stimulation, increase lysyl oxidase expression, inhibit matrix metalloproteinase 1(MMP1) activity, increase occludin production, increase filaggrin production, increase skin moisturization, and wherein nitric oxide synthase is inhibited, alpha 2A adrenergic receptor agonist activity is increased, Reduced oxidation, increased antioxidant capacity of the skin, inhibited COX-2 production, inhibited VEGF production, inhibited IL-6 production, inhibited IL-8 production, inhibited TNF- α production, increased collagen stimulation, increased expression of lysyl oxidase, inhibited MMP1 activity, increased occludin production, increased filaggrin production, and/or increased skin moisturization.

5. The method of claim 1, wherein the saccharide isomerate and/or the millettia extract is an aqueous extract.

6. The method of claim 1, wherein the saccharide isomerate comprises exopolysaccharides of vibrio alginolyticus belonging to the marine plankton family.

7. The method according to claim 1, wherein the extract of Artocarpus heterophyllus is an extract of leaves and stems of Artocarpus heterophyllus.

8. The method of claim 1, wherein the date extract is a water soluble extract from date seeds.

9. The method of claim 1, wherein the tea tree essential oil is an essential oil from the leaves of melaleuca alternifolia.

10. The method of claim 1, wherein the topical composition is a lotion, serum, gel lotion, or gel serum.

11. The method of claim 1, wherein the composition comprises an effective amount of date palm extract to increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of skin, inhibit cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) production, decrease tumor necrosis factor alpha (TNF-alpha) production, increase collagen stimulation, increase expression of lysyl oxidase, and/or inhibit the activity of matrix metalloproteinase 1(MMP 1);

an effective amount of tea tree essential oil to kill both follicular Demodex mites and sebaceous Demodex mites;

an effective amount of saccharide isomerate to reduce tumor necrosis factor alpha (TNF-alpha) production, inhibit nitric oxide synthase, increase occludin production, increase filaggrin production, and/or increase skin moisturization; and

an effective amount of Artocarpus heterophyllus extract to reduce the production of tumor necrosis factor alpha (TNF-alpha) and/or inhibit nitric oxide synthase.

12. A topical skin composition applied according to the method of claim 1, wherein the composition comprises date palm extract, tea tree essential oil, millettia extract, and saccharide isomers.

13. The topical composition of claim 12, wherein the topical composition comprises an effective amount of date palm extract, tea tree essential oil, Artocarpus heterophyllus extract, and saccharide isomerate to reduce rosacea, erythema, inflammation, transient or persistent erythema, telangiectasia, inflammatory papules and/or pustules, transient or persistent skin flushing, and/or proliferation of connective tissue.

14. The topical composition of claim 12, wherein the topical composition comprises an effective amount of date palm extract, tea tree essential oil, Artocarpus heterophyllus extract, and saccharide isomerate to inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, reduce oxidation, increase antioxidant capacity of the composition or skin, inhibit cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibiting the production of interleukin-6 (IL-6) and interleukin-8 (IL-8), reducing the production of tumor necrosis factor alpha (TNF-alpha), increasing collagen stimulation, increasing the expression of lysyl oxidase, inhibiting matrix metalloproteinase 1(MMP1) activity, increasing the production of occludin, increasing the production of filaggrin, and/or increasing skin moisturization.

15. The topical composition of claim 12, wherein the saccharide isomerate and/or the millettia extract is an aqueous extract.

16. The topical composition of claim 12, wherein the saccharide isomerate comprises exopolysaccharides of vibrio alginolyticus belonging to the marine plankton family.

17. The topical composition of claim 12, wherein the extract of Artocarpus heterophyllus is an extract of leaves and stems of Artocarpus heterophyllus.

18. The topical composition of claim 12, wherein the date extract is a water soluble extract from date seeds.

19. The topical composition of claim 12, wherein the tea tree essential oil is an essential oil from the leaves of melaleuca alternifolia.

20. The topical composition of claim 12, wherein the topical composition is a lotion, serum, gel lotion, or gel serum.

Background

A. Field of the invention

The present invention relates generally to topical skin care compositions that can reduce rosacea, erythema and/or skin inflammation and inhibit nitric oxide synthase. The combined ingredients may comprise date (Phoenix dactylifera) extract, tea tree essential oil, Artocarpus heterophyllus (Myrothamnus flabellifolia) extract and saccharide isomers.

B. Background of the invention

Rosacea is a common skin condition that affects millions of people and negatively impacts their quality of life. In the united states alone, an estimated about 1400 million people suffer from rosacea. Women over the age of 30 years with fair skin, particularly sunburned skin, are more likely to develop rosacea. Other risk factors include smoking, family history of rosacea, and keltt or scandinavia lineages. While women are more likely to develop rosacea, men are more likely to develop more severe conditions.

Rosacea causes redness and visible blood vessels in the face of a person, usually affecting the central third of the face, particularly the nose, of a person, and has redness of varying intensity over time. One of the four subtypes of rosacea is rosacea which is a type of erythematous vasodilation characterized by redness, flushing, and visible blood vessels. Typical facial redness associated with this type can persist in a person's face for a long period of time, with swelling of small blood vessels on the nose and cheeks, and become more pronounced upon onset. The second sub-type is papulopustular rosacea, characterized by redness, swelling, and acneiform eruptions. In some cases, swollen red bumps and papules can also contain pus and feel pain or heat when touched. The third subtype is rhinophyma-type rosacea, which is characterized by thickening of the skin and uneven texture of the skin, and in more severe cases can cause the rhinophyma, i.e., thickening of the skin over the nose, resulting in the nose appearing bulbous. Rosacea is the fourth subtype, and is the most severe form of rosacea, affecting one's eyes and causing dryness, irritation, eyelid swelling and redness, and the appearance of bumps that look like hordeolum.

Although the underlying cause of rosacea is unclear, the underlying process involves dilation of small blood vessels in the face. The ability of rosacea patients to inhibit facial inflammation, which may be caused by environmental factors such as sunburn, folliculitis (Demodex folliculorum), flushing, and specific medications, is often reduced by genetic factors. Rosacea tends to affect the "flush" areas of the face, more commonly among people who are prone to flushing. In addition, various factors are known to exacerbate the symptoms of rosacea. These factors include emotional factors (e.g., stress, fear, anxiety, embarrassment, depressed mood, etc.) as well as environmental factors (e.g., strong wind, changes in humidity, sunlight, and sunburn of the skin). Sports, alcohol drinking, smoking and spicy food are all common causes of exacerbating rosacea by increasing blood flow to the skin surface.

Others have attempted to make compositions and methods that reduce rosacea, skin inflammation, and erythema. Some have attempted to address multiple pathways involved in skin inflammation simultaneously by using multiple active ingredients in a single composition. For example, U.S. patent No. 8535738 and U.S. patent No. 9687517 teach several pathways that may be associated with inflammation and provide a list of ingredients that may address some of these pathways. However, only a few combinations of active ingredients were actually tested for compatibility and effectiveness. It is unclear whether the combination of the other ingredients therein is effective, may cause undesirable side effects, or may exacerbate the problems associated with skin inflammation.

Treatment of rosacea may also include the use of antibiotics, such as tetracycline, clindamycin, erythromycin and ivermycin (e.g., tetracycline, clindamycin, erythromycin)Cream 1%). See, e.g., U.S. patent No. 5972993 and U.S. patent publication No. 2015/0011489. Or, already will(Metronidazole 0.75% gel) is used as a topical composition that has limited therapeutic efficacy in treating papules and pustules associated with rosacea, but is ineffective in reducing redness, telangiectasia, or flushing of the skin. See, for example, U.S. patent No. 5972993. More recently, azelaic acid (e.g., NONE)15 percent of gel,20% cream) showed promise in treating mild to moderate papules and pustules by reducing the anti-inflammatory effects of reactive oxygen species. See Cole, Gary W., Rosacea, Medicine Neet (1/4.2019), https:// www.medicinenet.com/Rosacea/article. htm # Rosacea _ faces and Golgick H., and Layton, A. (2008), Azelaic acid 15% gel in the treatment of Rosacea, Expert Opinion on Pharmacotherapy,9(15), 2699-2706. DOI 10.1517/14656566.9.15.2699. Furthermore, the antiparasitic and anti-inflammatory properties of ivermectin have been reported to be effective in treating tumors, but the exact mechanism of action of ivermectin and its long-term effects are not known. Galderma, Soolantra (1/4/2019), https:// www.soolantra.com/about-rosacea-treatment.

Clear pox essence has also been used, but these are aesthetically unattractive as they can contain about 2% to 5% sulfur, which leaves an unpleasant odor. U.S. patent No. 5972993 teaches topical compositions for treating rosacea comprising an antioxidant selected from sulfur-containing compounds. While imidazole drugs (e.g., ketoconazole) have been shown to be useful in treating rosacea, the safety of continued long-term use of potent antimicrobials, antifungals, and antibacterials is unknown and may contribute to resistance. In the case of more severe rosacea, corticosteroid drugs and retinoids (e.g., isotretinoin) are used, but these drugs carry a greater risk of producing adverse side effects than antibiotics, thus limiting their long-term use.

Folliculosis, or uncontrolled infection by Demodex mites (both follicular Demodex brevis) a common epizootic parasite that parasitizes the pilosebaceous glands of the skin, is associated with a variety of skin disorders, including rosacea, as a cause of papulopustular skin lesions and perifollicular and inflammatory infiltrations. Once folliculitis affects the face, it spreads and develops easily on the eyelids because the skin in the periorbital area is not easily cleaned on a daily basis due to the surrounding protruding body parts such as the nose, eyebrows, and cheek areas. Demodex mite prevalence increases with age, and 84% to 100% of people in the 60-70 year old population become infected with Demodex mites. Tea tree essential oil (1% 4-terpene alcohol) has been shown to have a miticidal effect against demodex mites. Tighe et al (2013), Terphin-4-ol is the most active ingredient of tea tree oil to kill Demodex mite, Translational Vision Science & Technology,2(7), 1-8. DOI 10.1167/tvst.2.7.2.

Many of these attempts to treat rosacea have been ineffective, addressing only one or more of the undesirable effects of rosacea, inflammation, and erythema, or causing unacceptable side effects in themselves, such as skin irritation or allergic reactions. Furthermore, not every effective composition is suitable for every skin type. Thus, there is a need for new products that effectively reduce rosacea, erythema and inflammation of the skin.

Disclosure of Invention

The present inventors have discovered a solution to at least some of the problems associated with rosacea, erythema and/or inflammation of the skin. The solution is based on a combination of ingredients that may include date extract, tea tree essential oil, millettia extract and saccharide isomers. The combination can be used to make a topical composition effective in reducing rosacea, erythema and/or skin inflammation and reducing nitric oxide synthase activity. In some cases, date extract was demonstrated to increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of the composition or skin, inhibit cyclooxygenase-2 (COX-2), Vascular Endothelial Growth Factor (VEGF), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF-alpha) production, increase collagen stimulation and lysyl oxidase expression, and inhibit the activity of matrix metalloproteinase 1(MMP 1). In some cases, saccharide isomerate is demonstrated to reduce tumor necrosis factor alpha (TNF- α) production, inhibit nitric oxide synthase, increase production of occludin and filaggrin, and enhance skin moisturization. In some particular cases, extracts of Artocarpus heterophyllus have been shown to reduce TNF- α production and inhibit nitric oxide synthase. In some cases, the extract is an aqueous extract. In some cases, other solvents such as alcohols, glycols, hydro-alcohols, and/or hydro-glycol extracts may be used.

In some aspects, topical compositions are disclosed. In some aspects, the topical composition comprises any one, any combination, or all of date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate. In some cases, the topical composition comprises an effective amount of date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate to reduce rosacea, erythema, and/or inflammation. In some cases, the topical composition comprises an effective amount of date extract, tea tree essential oil, melaleuca extract, and saccharide isomerate to reduce transient or persistent erythema, telangiectasia, inflammatory papules, pustules, transient flushing of the skin, persistent flushing of the skin, and/or hyperplasia of connective tissue. In some cases, the topical composition comprises an effective amount of date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate to inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of the composition or the skin, inhibit COX-2 production, inhibit VEGF production, inhibit IL-6 production, inhibit IL-8 production, decrease TNF-alpha production, increase collagen stimulation, increase expression of lysyl oxidase, inhibit MMP1 activity, increase occludin production, increase filaggrin production, and/or increase skin moisturization. In some cases, the topical composition includes an effective amount of date palm extract to increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of the composition or skin, inhibit COX-2 production, inhibit VEGF production, inhibit IL-6 production, inhibit IL-8 production, decrease TNF-alpha production, increase collagen stimulation, increase expression of lysyl oxidase, and/or inhibit the activity of MMP 1. In some cases, the topical composition comprises an effective amount of tea tree essential oil to kill both follicular demodex and sebaceous demodex. In some cases, the topical composition includes an effective amount of saccharide isomerate to reduce TNF- α production, inhibit nitric oxide synthase, increase occludin production, increase filaggrin production, and/or increase skin moisturization. In some cases, the topical composition comprises an effective amount of an extract of Artocarpus heterophyllus to reduce TNF- α production and/or inhibit nitric oxide synthase.

In some cases, the topical composition comprises water. In some cases, the saccharide isomerate and/or the millettia extract is an aqueous extract. Aqueous extracts mean that aqueous solutions can be used as extractants or solvents to obtain the extract. In addition to water, in some cases, the aqueous solution may include alcohols, glycols, or combinations thereof. The aqueous extract may be in liquid form or in powder form. The saccharide isomerate may comprise exopolysaccharides of Vibrio alginolyticus (Vibrio alginolyticus) belonging to the marine plankton family. The extract of Artocarpus heterophyllus can be extract of leaves and stems of Artocarpus heterophyllus. The date extract may be a water soluble extract from date seeds. The tea tree essential oil may be an essential oil from the leaves of Melaleuca alternifolia (Melaleuca alternifolia).

The topical compositions disclosed herein may also comprise one or more than one ingredient described herein. For example, the composition may comprise one or more additional ingredients selected from one or more conditioning agents, humectants, pH modifying agents, structuring agents, inorganic salts, and preservatives. In some cases, the topical composition further comprises water. The amount of ingredients in the composition may vary (e.g., the amount may be as low as 0.000001 wt% to as high as 98 wt%, or any range therebetween). In some cases, the topical composition is a lotion, serum, gel lotion, or gel serum.

Methods of using the compositions disclosed herein are also disclosed. In some aspects, methods of improving the condition or appearance of skin are disclosed, comprising applying any one of the compositions disclosed herein to skin in need thereof. In one aspect, any one of the compositions disclosed herein is applied to the skin and the composition is left on the skin or removed from the skin after a period of time. In some aspects, the compositions disclosed herein are used to treat and/or reduce rosacea. In some aspects, the compositions disclosed herein are used to treat and/or reduce erythema. In some aspects, the compositions disclosed herein are used to treat and/or reduce inflammation. In another aspect, the compositions disclosed herein are used to inhibit or reduce nitric oxide synthase activity. In some aspects, the compositions disclosed herein are used to increase alpha 2A adrenergic receptor agonist activity, reduce oxidation, inhibit COX-2 production, inhibit VEGF production, inhibit IL-6 production, inhibit IL-8 production, reduce TNF-alpha production, increase collagen stimulation, increase expression of lysyl oxidase, inhibit MMP1 activity, kill demodex folliculorum and/or demodex sebaceus, inhibit nitric oxide synthase, increase occludin production, increase filaggrin production, and/or increase skin moisturization. In some cases, the topical compositions disclosed herein comprise an effective amount of date extract and are used to increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, inhibit COX-2 production, inhibit VEGF production, inhibit IL-6 production, inhibit IL-8 production, decrease TNF-alpha production, increase collagen stimulation, increase expression of lysyl oxidase, and/or inhibit MMP1 activity. In some cases, the compositions disclosed herein comprise an effective amount of tea tree essential oil and are used to kill both follicular demodex and sebaceous demodex. In some cases, the compositions disclosed herein comprise an effective amount of saccharide isomerate and are useful for reducing TNF- α production, inhibiting nitric oxide synthase, increasing occludin production, increasing filaggrin production, and/or increasing skin moisturization. In some cases, the compositions disclosed herein comprise an effective amount of an extract of Artocarpus heterophyllus and are used to reduce the production of tumor necrosis factor alpha (TNF-alpha) and/or inhibit nitric oxide synthase. In some aspects, the method comprises applying any of the topical compositions described herein to the skin. In some aspects, the method comprises applying the composition to facial skin. In some cases, the method comprises applying the composition to red skin, rosacea skin, erythematous skin, and/or inflamed skin.

In some aspects, the compositions of the present invention are formulated as topical skin compositions. The compositions may have a dermatologically acceptable carrier or vehicle for the compounds and extracts. The composition may further comprise a humectant or wetting agent, a surfactant, a silicone-containing compound, a UV agent, an oil, and/or other ingredients identified herein or known in the art. The composition can be a mask, lotion, cream, gel, serum, emulsion (e.g., oil-in-water, water-in-oil, silicone-in-water, water-in-oil-in-water, oil-in-water-in-oil, oil-in-water-in-silicone, etc.), solution (e.g., aqueous or hydroalcoholic solution), anhydrous base (e.g., lipstick or powder), ointment, milk, paste, aerosol, solid form, eye gel, gel serum, gel emulsion, etc. The composition may be formulated for topical skin application, applied at least 1,2, 3, 4,5, 6, 7, or more than 7 times per day during use. In other aspects of the invention, the compositions may be storage stable or color stable, or both. It is also contemplated that the viscosity of the composition can be selected to achieve a desired result, for example, the viscosity of such composition can be from about 1cp to well over 1 million cp, or any range or integer derivable therein, depending on the type of composition desired (e.g., 2cp, 3cp, 4cp, 5cp, 6cp, 7cp, 8cp, 9cp, 10cp, 20cp, 30cp, 40cp, 50cp, 60cp, 70cp, 80cp, 90cp, 100cp, 200cp, 300cp, 400cp, 500cp, 600cp, 700cp, 800cp, 900cp, 1000cp, 2000cp, 3000cp, 4000cp, 5000cp, 6000cp, 7000cp, 8000cp, 9000cp, 10000cp, 20000cp, 30000cp, 40000cp, 50000cp, 60000cp, 70000cp, 80000cp, 90000cp, 900000cp, 400000cp, 500000cp, 80cp, measured at 2.5rpm on a brookfield viscometer at 25 ℃.(s), 1000000cp, 2000000cp, 3000000cp, 4000000cp, 5000000cp, 10000000cp, etc.).

In a non-limiting aspect, the pH of the composition can be from about 6 to about 9. In other aspects, the pH can be 1,2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, or 14. The composition may comprise triglycerides. Non-limiting examples include short, medium and long chain triglycerides. In certain aspects, the triglyceride is a medium chain triglyceride (e.g., caprylic capric triglyceride). The composition may also comprise a preservative. Non-limiting examples of preservatives include methyl paraben, propyl paraben or a mixture of methyl paraben and propyl paraben. In some embodiments, the composition is free of parabens.

The compositions of the present invention may have UVA and UVB absorption properties. The composition may have a Sun Protection Factor (SPF) of 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or greater than 60, or any integer or derivative thereof. The composition may be a sunscreen lotion, sunscreen spray or sunscreen cream.

The compositions of the present invention may further comprise any one, any combination, or all of the following additional ingredients: water, chelating agents, humectants, preservatives, thickeners, silicone-containing compounds, essential oils, structuring agents, vitamins, pharmaceutical ingredients or antioxidants, or any combination of these ingredients or mixtures of these ingredients. In certain aspects, the composition can comprise at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, or all of these additional ingredients identified in the foregoing. Non-limiting examples of these additional ingredients are identified throughout this specification and are incorporated by reference into this section. Amounts of such ingredients may range from 0.0001% to 99.9% by weight or volume of the composition, or any integer or range between as disclosed in other sections of this specification, which are incorporated herein by reference.

Kits comprising the compositions of the invention are also contemplated. In certain embodiments, the composition is contained in a container. The container may be a bottle, dispenser or pack/bag. The container may dispense a predetermined amount of the composition. In particular aspects, the composition is dispensed in the form of a spray, mist, bolus or liquid. The container may contain indicia on its surface. The indicia may be words, abbreviations, pictures or symbols.

It is also contemplated that the compositions disclosed throughout this specification may be used as leave-on or rinse-off compositions. For example, the resident composition may be topically applied to the skin and held on the skin for a period of time (e.g., at least 5 minutes, at least 6 minutes, at least 7 minutes, at least 8 minutes, at least 9 minutes, at least 10 minutes, at least 20 minutes, or at least 30 minutes, or at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, or at least 24 hours, or overnight or entire day). Alternatively, the rinse-off composition may be a product that is to be applied to the skin and then removed or washed off the skin (e.g., with water) over a period of time, such as less than 5 minutes, less than 4 minutes, less than 3 minutes, less than 2 minutes, or less than 1 minute. Examples of rinse-off compositions may be skin cleansers, shampoos, conditioners or soaps. Examples of leave-on compositions may be skin creams, sunscreens, face masks, night creams or day creams.

It is contemplated that any embodiment discussed in this specification can be implemented for any method or composition of the invention and vice versa. Furthermore, the compositions of the present invention may be used to carry out the methods of the present invention.

In one embodiment, the compositions of the present invention may be pharmaceutically or cosmetically acceptable, or may have pleasant tactile properties. "pharmaceutically acceptable", "cosmetically acceptable", and/or "pleasant tactile properties" describe compositions having specific tactile properties that are pleasant to the skin (e.g., less dilute or less oily compositions, compositions having a silky texture, non-tacky or sticky compositions, etc.). Pharmaceutically acceptable or cosmetically acceptable may also relate to the creaminess or lubrication properties of the composition, or the moisturizing properties of the composition.

Products comprising the compositions of the present invention are also contemplated. In a non-limiting aspect, the product can be a cosmetic product. The cosmetic product may be a cosmetic product as described elsewhere in this specification, or a cosmetic product known to those skilled in the art. Non-limiting examples of products include skin creams, lotions, emollients, gels, lotions, foundations, night creams, lipsticks, cleansers, toners, sunscreens, masks, anti-aging products, deodorants, antiperspirants, perfumes, colognes, and the like.

The following embodiments 1 to 38 of the present invention are also disclosed. Embodiment 1 is a method of treating skin comprising topically applying to the skin an effective amount of a topical composition comprising date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate, wherein the skin is treated. Embodiment 2 is the method of embodiment 1, wherein the skin is treated to reduce rosacea, erythema and/or inflammation, and wherein the rosacea, erythema and/or inflammation is reduced. Embodiment 3 is the method of embodiments 1-2, wherein the skin is treated to reduce temporary or persistent erythema, telangiectasia, inflammatory papules and/or pustules, temporary or persistent skin flushing, and/or proliferation of connective tissue. Embodiment 4 is the method of any one of embodiments 1 to 3 wherein the skin is treated to inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, reduce oxidation, increase antioxidant capacity of the skin, reduce cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) production, reduce tumor necrosis factor alpha (TNF-alpha) production, increase collagen stimulation, increase lysyl oxidase expression, inhibit matrix metalloproteinase 1(MMP1) activity, increase occludin production, increase filaggrin production, increase skin moisturization, and wherein nitric oxide synthase is inhibited, alpha 2A adrenergic receptor agonist activity is increased, a method of treating skin to inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, decrease tumor necrosis factor alpha (TNF-alpha) production, decrease production of tumor necrosis factor alpha (TNF-alpha-2) production, decrease production of tumor necrosis factor alpha-adrenergic receptor agonist activity, decrease skin moisturization, and/or inflammation, Reduced oxidation, increased antioxidant capacity of the skin, inhibited COX-2 production, inhibited VEGF production, inhibited IL-6 production, inhibited IL-8 production, inhibited TNF- α production, increased collagen stimulation, increased expression of lysyl oxidase, inhibited MMP1 activity, increased occludin production, increased filaggrin production, and/or increased skin moisturization. Embodiment 5 is the method of any one of embodiments 1 to 4, wherein the topical composition further comprises water. Embodiment 6 is the method of any one of embodiments 1 to 5, wherein the saccharide isomers and/or the millettia extract are aqueous extracts. Embodiment 7 is the method of embodiment 6, wherein the aqueous extract is in liquid form. Embodiment 8 is the method of embodiment 6, wherein the aqueous extract is in powder form. Embodiment 9 is the method of any one of embodiments 1 to 8, wherein the saccharide isomerate comprises exopolysaccharides of vibrio alginolyticus belonging to the marine plankton family. Embodiment 10 the method of any one of embodiments 1 to 9 wherein the extract of millettia speciosa is an extract of the leaves and stems of millettia speciosa. Embodiment 11 is the method of any one of embodiments 1 to 10, wherein the date extract is a water soluble extract from date seeds. Embodiment 12 is the method of any one of embodiments 1 to 11, wherein the tea tree essential oil is an essential oil from the leaves of melaleuca alternifolia. Embodiment 13 is the method of any one of embodiments 1 to 12, wherein the topical composition is a lotion, serum, gel lotion, or gel serum. Embodiment 14 is the method of any one of embodiments 1 to 13, wherein the topical composition is an oil-in-water or water-in-oil emulsion. Embodiment 15 is the method of any one of embodiments 1 to 14, wherein the composition is applied to facial skin. Embodiment 16 is the method of any one of embodiments 1 to 15, wherein the composition comprises an effective amount of date palm extract to increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of skin, inhibit cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) production, decrease tumor necrosis factor alpha (TNF-alpha) production, increase collagen stimulation, increase lysyl oxidase expression, and/or inhibit matrix metalloproteinase 1(MMP1) activity. Embodiment 17 is the method of any one of embodiments 1 to 16, wherein the composition comprises an effective amount of tea tree essential oil to kill both follicular demodex and sebaceous demodex. Embodiment 18 is the method of any one of embodiments 1 to 15, wherein the composition comprises an effective amount of saccharide isomerate to reduce tumor necrosis factor alpha (TNF-alpha) production, inhibit nitric oxide synthase, increase occludin production, increase filaggrin production, and/or increase skin moisturization. Embodiment 19 is the method of any one of embodiments 1 to 15, wherein the composition comprises an effective amount of an extract of Artocarpus heterophyllus to reduce the production of tumor necrosis factor alpha (TNF- α) and/or inhibit nitric oxide synthase. Embodiment 20 is a topical skin composition comprising date extract, tea tree essential oil, millettia extract, and saccharide isomers. Embodiment 21 is the composition of embodiment 20, wherein the topical composition comprises an effective amount of date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate to reduce rosacea, erythema, and/or inflammation. Embodiment 22 is the composition of any one of embodiments 20 to 21, wherein the topical composition comprises an effective amount of date extract, tea tree essential oil, melaleuca extract, and saccharide isomerate to reduce temporary or persistent erythema, telangiectasia, inflammatory papules and/or pustules, temporary or persistent skin flushing, and/or proliferation of connective tissue. Embodiment 23 is the composition of any one of embodiments 20 to 22, wherein the topical composition comprises an effective amount of date extract, tea tree essential oil, millettia extract, and saccharide isomerate to inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of the composition or skin, inhibit cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibiting the production of interleukin-6 (IL-6) and interleukin-8 (IL-8), reducing the production of tumor necrosis factor alpha (TNF-alpha), increasing collagen stimulation, increasing the expression of lysyl oxidase, inhibiting matrix metalloproteinase 1(MMP1) activity, increasing the production of occludin, increasing the production of filaggrin, and/or increasing skin moisturization. Embodiment 24 is the composition of any one of embodiments 20 to 23, wherein the saccharide isomers and/or the millettia extract are aqueous extracts. Embodiment 25 is the topical composition of embodiment 24, wherein the aqueous extract is in liquid form. Embodiment 26 is the topical composition of embodiment 24, wherein the aqueous extract is in powder form. Embodiment 27 is the composition of any one of embodiments 20 to 26, wherein the saccharide isomers comprise exopolysaccharides of vibrio alginolyticus belonging to the marine plankton family. Embodiment 28 the composition of any one of embodiments 20 to 27 wherein the extract of millettia speciosa is an extract of the leaves and stems of millettia speciosa. Embodiment 29 is the composition of any one of embodiments 20 to 28, wherein the date extract is a water soluble extract from date seed. Embodiment 30 is the composition of any one of embodiments 20 to 29, wherein the tea tree essential oil is an essential oil from the leaves of melaleuca alternifolia. Embodiment 31 is the composition of any one of embodiments 20 to 30, wherein the topical composition further comprises water. Embodiment 32 is the composition of any one of embodiments 20 to 31, wherein the topical composition is a lotion, serum, gel lotion, or gel serum. Embodiment 33 is the composition of any one of embodiments 20 to 32, wherein the topical composition is an oil-in-water or water-in-oil emulsion. Embodiment 34 is the composition of any one of embodiments 20 to 33, wherein the topical composition is formulated for application to facial skin. Embodiment 35 is the composition of any one of embodiments 20 to 34, wherein the topical composition comprises an effective amount of phoenix dactylifera extract to increase alpha 2A adrenergic receptor agonist activity, decrease oxidation, increase antioxidant capacity of the composition or skin, inhibit cyclooxygenase-2 (COX-2) production, inhibit Vascular Endothelial Growth Factor (VEGF) production, inhibit interleukin-6 (IL-6) and interleukin-8 (IL-8) production, decrease tumor necrosis factor alpha (TNF-alpha) production, increase collagen stimulation, increase lysyl oxidase expression, and/or inhibit matrix metalloproteinase 1(MMP1) activity. Embodiment 36 is the composition of any one of embodiments 20 to 35, wherein the topical composition comprises an effective amount of tea tree essential oil to kill both follicular demodex and sebaceous demodex. Embodiment 37 is the composition of any one of embodiments 20 to 36, wherein the topical composition comprises an effective amount of a saccharide isomer to reduce tumor necrosis factor alpha (TNF-alpha) production, inhibit nitric oxide synthase, increase occludin production, increase filaggrin production, and/or increase skin moisturization. Embodiment 38 is the composition of any one of embodiments 20 to 37, wherein the topical composition comprises an effective amount of an extract of Artocarpus heterophyllus to reduce the production of tumor necrosis factor alpha (TNF- α) and/or inhibit nitric oxide synthase.

By "topical application" is meant that the composition is applied or spread onto the surface of the lips or keratinous tissue. "topical skin compositions" include compositions suitable for topical application to the skin and/or keratinous tissue. Such compositions are generally dermatologically acceptable in that they do not have abnormal toxicity, incompatibility, instability, allergic response, and the like when applied to the skin and/or keratinous tissue. The topical skin care compositions of the present invention can have a viscosity selected to avoid significant dripping or pooling after application to the skin and/or keratinous tissue.

"rosacea" includes, but is not limited to, rosacea, papulopustular rosacea, rhinophyma-type rosacea, and ocular rosacea. Rosacea symptoms that may be treated by the present invention include, but are not limited to, any one, combination, or all of the following: redness, transient or persistent skin flushing, visible appearance of blood vessels (particularly in the nose or cheek area of the face), swelling, swollen red bumps, and papules that may contain pus, transient or persistent erythema, inflammation, skin thickening, uneven skin texture, dry skin, skin irritation, telangiectasia, inflammatory papules and/or pustules, and/or hyperplasia of connective tissue.

"keratinous tissue" includes a stratum corneum-containing layer that is the outermost protective covering for mammals, including but not limited to lips, skin, hair, and nails.

The term "about" or "approximately" is defined as being close as understood by one of ordinary skill in the art. In a non-limiting embodiment, the term is defined as within 10%, preferably within 5%, more preferably within 1%, and most preferably within 0.5%.

The term "substantially" and variations thereof refer to a range within 10%, within 5%, within 1%, or within 0.5%.

The terms "inhibit" or "reduce" or any variation of these terms includes any measurable reduction or complete inhibition to achieve the intended result. The terms "promote" or "increase" or any variant of these terms include any measurable increase or production of a protein or molecule (e.g., a matrix protein such as fibronectin, laminin, collagen or elastin, or a molecule such as hyaluronic acid) in order to achieve the intended result.

As used in this specification and/or in the claims, the term "effective" means suitable for achieving a desired, expected, or expected result.

The use of quantitative terms preceded by the term "a" or "an" when used in the claims and/or the specification with the term "comprising", "including", "having" or "containing" or any variation of these terms may mean "one", but it also conforms to the meaning of "one or more", "at least one" and "one or more".

As used in this specification and claims, the words "comprise," "have," "include," or "contain" are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

The compositions and methods of use may "comprise," consist essentially of, "or" consist of any of the ingredients or steps disclosed throughout this specification. With respect to the phrase "consisting essentially of … …," the basic and novel properties of the compositions and methods of the present invention are that the compositions contain date extract, tea tree essential oil, millettia extract, and saccharide isomers. Another novel property of the compositions and methods is the use of the compositions to treat, reduce and/or prevent, in whole or in part, rosacea, erythema and/or inflammation of the skin, or a symptom or cause thereof.

Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the embodiments, while indicating specific embodiments of the invention, are given by way of illustration only. In addition, it is contemplated that variations and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

Detailed Description

As mentioned above, the present invention provides a solution to the problems associated with rosacea, erythema of the skin, and/or inflammation of the skin. The solution is based on the use of a combination of date extract, tea tree essential oil, millettia extract and saccharide isomerate to reduce rosacea, skin erythema and/or skin inflammation. As illustrated in the examples, in a non-limiting manner, the combination has been shown to reduce rosacea, skin erythema and inflammation, as well as inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, reduce oxidation, inhibit COX-2 production, inhibit VEGF production, inhibit IL-6 production, inhibit IL-8 production, reduce TNF-alpha production, increase collagen stimulation, increase lysyl oxidase expression, inhibit MMP1 activity, reduce occludin production, increase filaggrin production, and/or increase skin moisturization.

These and other non-limiting aspects of the invention are described in the following sections.

A. Active ingredient

The present invention is premised on the identification of a combination of active ingredients-date extract, tea tree essential oil, millettia extract and saccharide isomerate-useful for reducing acne rosacea, erythema and/or inflammation of the skin, and inhibiting nitric oxide synthase, increasing alpha 2A adrenergic receptor agonist activity, reducing oxidation, increasing antioxidant capacity of the composition or skin, inhibiting COX-2 production, inhibiting VEGF production, inhibiting IL-6 production, inhibiting IL-8 production, reducing TNF-alpha production, increasing collagen stimulation, increasing lysyl oxidase expression, inhibiting MMP1 activity, increasing occludin production, increasing filaggrin production and/or increasing skin moisturization.

The combination of ingredients can be used in different products to treat a variety of skin conditions. By way of non-limiting example, the combination of ingredients may be formulated as an emulsion (e.g., oil-in-water, water-in-oil), a gel, an essence, a gel emulsion, a gel essence, a skin lotion, a mask, or a body lotion.

Saccharide isomerate is an exopolysaccharide synthesized by microorganisms called vibrio alginolyticus, which belong to the marine plankton family. In some cases, saccharide isomers are commercially available. In some cases, saccharide isomers are available under the trade name Benoiderm from Barnet Products. In some cases, the saccharide isomer may be provided by extraction from vibrio alginolyticus using an aqueous extraction solvent or an alcoholic extraction solvent. In other cases, the extraction solvent may be aqueous. In some cases, the extract may be in liquid form. In some cases, the extract may be in powder form.

The extract of millettia is an extract of millettia, also known as resuscitation plant, which is a flowering plant native to south africa. In some cases, the millettia extract is commercially available. In some cases, the extract of Artocarpus heterophyllus may be under the trade name Artocarpus heterophyllusSupplied by Rahn. In some cases, the extract may be an aqueous or alcoholic extract. In some cases, the extract may be an aqueous extract. In some cases, the extract is an extract of the whole plant or one or more parts of the plant. In some cases, the extract is an extract of a leaf or stem of the plant. In some cases, the extract may be in liquid form. In some cases, the extract may be in powder form.

Tea tree essential oil, also known as cajeput essential oil or tea tree oil, is an essential oil derived from the leaves of the melaleuca alternifolia of the tea tree. In some cases, tea tree essential oil is commercially available. In some cases, tea tree essential oil may be sold under the tradename MELAFRESH^TMT96 was supplied by Southern Cross Botanicals.

The date extract is an extract of date, also known as date palm, a flowering plant species of Arecaceae, a family palmaceae. In some cases, date is commercially available. In some cases, date extracts may be tradenameProvided by IBR ltd. In some cases, the extract may be a water-soluble extract from date seed. In some cases, the extract may be an aqueous or alcoholic extract. In some cases, the extract may be an aqueous extract. In some cases, the extract is an extract of the whole plant or one or more parts of the plant. In some cases, the extract is an extract of a whole plant. In some cases, the extract is an extract of seeds and/or fruits。

The extracts described herein can be extracts prepared by extraction methods known in the art and combinations thereof. Non-limiting examples of extraction methods include the use of liquid-liquid extraction, solid phase extraction, water extraction, ethyl acetate extraction, alcohol extraction, acetone extraction, oil extraction, supercritical carbon dioxide extraction, heat extraction, pressure drop extraction, ultrasonic extraction, and the like. The extract can be liquid, solid, dried liquid, re-suspended solid, etc. In particular instances, the extract is an aqueous extract.

B. Amount of ingredient

It is contemplated that the compositions of the present invention may contain any amount of the ingredients discussed in this specification. The composition may also contain any number of combinations of additional ingredients (e.g., pigments or additional cosmetic or pharmaceutical ingredients) described throughout this specification. The concentration of the ingredients in the composition may vary. For example, in non-limiting embodiments, the composition may comprise, consist essentially of, or consist of, in its final form: for example, at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.0039%, 0.0040.0040%, 0051%, 0040.0043%, 0.0040.0040.0070%, 0060.0075%, 0.0070.0070.0070.0060.0060.0075%, 0.0070.0070.0070.0060.0075%, 0.0070.1%, 0.0040.0050.0075%, 0.0070.1%, 0.0050.0070.1%, 0.0040.0040.0050.0075%, 0.8%, 0.0070.0040.8%, 0.0040.0050.0070.8%, 0.0040.0050.8%, 0.8%, 0.0040.8%, 0.0040%, 0.0040.0040%, 0.0040.8%, 0.0040.0050.0070.8%, 0.0040.1%, 0.0050.0050.0050.8%, 0.8%, 0.0050.0050.8%, 0.0070.8%, 0.0070.0050.1%, 0.0050%, 0.0050.1%, 0.0050.0070.0050.0050.1%, 0.0070.1%, 0.0070.0050.1%, 0.1%, 0.0070.0070.1%, 0.0070.1%, 0.0070.0050.0070.1%, 0.1%, 0.0070.1%, 0.0050.0070.0070.0050.0050.0070.0050.0050.0050.0050.0050.0050.0075%, 0%, 0.0075%, 0.1%, 0.0070.0070.0070.0070.0070.0050.0070.0070.0070.1%, 0.0070.1%, 0.1%, 0.0070.1%, 0.0050.1%, 0.0050.0070.0040.1%, 0.1%, 0.0070.1%, 0.1%, 0.0070.1%, 0.1%, 0.0070.0050.1%, 0.1%, 0.0070.0070.1%, 0.0040.1%, 0.0040.0050.0070.0050.0070.0040.0070.0070.0070.1%, 0.1%, 0.0070.0070.1%, 0.1%, 0.0070.0070.0070.0070.0070.1%, 0.1%, 0.0040.0070.1%, 0.0070.1%, 0.0070, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%, 0.0675%, 0.0700%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0800.0800.085%, 0.0850%, 0.080.080%, 0.0900%, 0.9%, 0.460%, 0.9%, 9%, 9.590.9%, 0.9%, 9%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 0%, 0.9%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6.6%, 6.7%, 7.9.7%, 7.9%, 8%, 7.9.9%, 7.9%, 7%, 8%, 7.9.9%, 7%, 7.9%, 8%, 7.9%, 7%, 7.9%, 8%, 7.9%, 7%, 8%, 7.9%, 7.9.9%, 7%, 8%, 7%, 8%, 7.9%, 7%, 8.8.8%, 7%, 7.9%, 7.8%, 7.8.8%, 7%, 7.9%, 7%, 7.8%, 7%, 7.8%, 7.8.8%, 7.9%, 7%, 7.8.8.8%, 7%, 7.0%, 7.9%, 7%, 7.8.8%, 7.9%, 7%, 7.8.8%, 7%, 7.9%, 8%, 8.9%, 7.9%, 7%, 7.9%, 7%, 8%, 7%, 7.8%, 8%, 7%, 7.9%, 8%, 7%, 7.9%, 7%, 7.8.9%, 7%, 8%, 7.9%, 7%, 7.9%, 7%, 7.9.9% or 8.9%, 7.9% or 8.9%, 7%, 7.9%, 7%, 7.9.9.9%, 7%, 7.9.9% or 8% or 8.9% or 8% or 6.9% or 8.9% or 8% or more, 9.9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or any range derivable therein of at least one of the ingredients mentioned throughout the specification and claims. In a non-limiting aspect, the percentages can be calculated by weight or volume of the entire composition. One of ordinary skill in the art will appreciate that the concentration in a given composition can vary depending on the addition, substitution, and/or subtraction of ingredients.

C. Carrier

The compositions of the present invention may comprise or be incorporated into all types of carriers and vehicles. The carrier or vehicle may be a pharmaceutically or dermatologically acceptable carrier or vehicle. Non-limiting examples of carriers and vehicles include water, glycerin, alcohols, oils, silicon-containing compounds, silicone compounds, and waxes. Variants and other suitable vectors will be apparent to the skilled artisan and are suitable for use in the present invention. In certain aspects, the concentrations and combinations of compounds, ingredients, and agents can be selected such that the compositions are chemically compatible and do not form complexes that precipitate out of the finished product.

D. Structure of the product

The compositions of the present invention may be configured or formulated in a variety of different forms. Non-limiting examples include emulsions (e.g., water-in-oil-in-water, silicone-in-water, water-in-silicone, oil-in-water-in-oil, oil-in-water-in-silicone emulsions), creams, lotions, solutions (aqueous or hydroalcoholic), anhydrous bases (e.g., lipsticks and loose powders), gels, masks, exfoliates, and ointments. Variations and other configurations will be apparent to the skilled artisan and are applicable to the present invention.

E. Additional ingredients

In addition to the combination of ingredients disclosed by the inventors, the composition may also contain additional ingredients, such as cosmetic ingredients and pharmaceutical active ingredients. Non-limiting examples of these additional components are described in the subsections that follow.

1. Cosmetic composition

CTFA international cosmetic ingredient dictionaries and manuals (2004 and 2008) describe a variety of non-limiting cosmetic ingredients that may be used in the context of the present invention. Examples of these ingredient classes include: fragrances (artificial and natural; e.g., gluconic acid, phenoxyethanol, and triethanolamine), dyes and coloring ingredients (e.g., blue No. 1, lake blue No. 1, Red No. 40, titanium dioxide, D & C blue No. 4, D & C Green No. 5, D & C orange No. 4, D & C Red No. 17, D & C Red No. 33, D & C Violet No. 2, D & C yellow No. 10, and D & C yellow No. 11), flavorants/fragrances (e.g., Stevia (Stevia rebaudiana) extract and menthol), adsorbents, lubricants, solvents, moisturizers (including, e.g., emollients, humectants, film formers, occlusive agents, and agents that affect the natural moisturizing mechanism of the skin), waterproofing agents, ultraviolet absorbers (physical and chemical absorbers, e.g., p-aminobenzoic acid ("PABA") and corresponding PABA derivatives, titanium dioxide, zinc oxide, and the like), Essential oils, vitamins (e.g., A, B, C, D, E and K), trace metals (e.g., zinc, calcium, and selenium), anti-irritants (e.g., steroids and non-steroidal anti-inflammatory drugs), plant extracts (e.g., Aloe vera (Aloe vera), chamomile, cucumber extract, Ginkgo biloba (Ginko biloba), ginseng and rosemary), antimicrobials, antioxidants (e.g., BHT and tocopherol), chelating agents (e.g., disodium EDTA and tetrasodium EDTA), preservatives (e.g., methyl paraben and propyl paraben), pH adjusters (e.g., sodium hydroxide and citric acid), adsorbents (e.g., aluminum starch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrin, talc, and zeolite), skin bleaching and lightening agents (e.g., hydroquinone and niacinamide lactate), humectants (e.g., sorbitol, urea, methyl gluceth-20, sodium and sodium, Isomerose and mannitol), keratolytic agents, water-blocking agents (e.g., sodium magnesium hydroxide/aluminum stearate), skin conditioning agents (e.g., aloe vera extract, allantoin, bisabolol, ceramide, polydimethylsiloxane, hyaluronic acid, biosaccharide gum-1, ethylhexylglycerin, pentylene glycol, hydrogenated polydecene, octyldodecyl oleate, and dipotassium glycyrrhizinate). Non-limiting examples of some of these ingredients are provided in the subsections below.

a. UV absorbing and/or reflecting agents

Uv absorbing and/or reflecting agents that may be used in combination with the compositions of the present invention include chemical and physical sunscreen substances. Non-limiting examples of chemical sunscreen substances that may be used include p-aminobenzoic acid (PABA), PABA esters (PABA glycerides, p-dimethylaminobenzoate pentyl and p-dimethylaminobenzoate octyl), PABA butyl esters, PABA ethyl esters, dihydroxypropanol PABA ethyl esters, benzophenones (oxybenzone, sulisobenzone, benzophenone, and benzophenone-1 to benzophenone-12), cinnamates (octyl methoxycinnamate (Octinoxate)), isoamyl p-methoxycinnamate, octyl methoxycinnamate, cinoxate, methyl diisopropylcinnamate, DEA methoxycinnamate, ethyl diisopropylcinnamate, dimethoxycinnamate glyceryl caprylate, and ethylmethoxycinnamate), cinnamates, salicylates (pimozo, benzyl salicylate, ethylene glycol salicylate, isopropylbenzyl salicylate, and the like), Anthranilate, ethyl urocanate, primisulfite, octyl salicylate, dibenzoylmethane derivatives (e.g. avobenzone), octocrylene, octyl triazone, galloylgallate trioleate, glyceryl aminobenzoate, dihydroxyacetone-containing lawsone, ethylhexyl triazone, dioctylbutylaminotriazinone, benzylidene malonate polysiloxanes, p-xylylene-dibornene sulfonic acid, phenylbisbenzimidazole tetrasulfonate disodium, diethylaminohydroxybenzoyl hexyl benzoate, bisdiethylaminohydroxybenzoyl benzoate, bisbenzoxazolyl phenylethylhexyl imino triazine, cresoltrazol trisiloxane, methylenebisbenzotriazolyl tetramethylbutyl phenol and bisethylhexyloxyphenol methoxyphenyl triazine, 4-methylbenzylidenecamphene and 4-methoxycinnamic acid isoamyl ester. Non-limiting examples of physical sunscreen substances include kaolin, talc, petrolatum, and metal oxides (e.g., titanium dioxide and zinc oxide).

b. Moisture-retaining agent

Non-limiting examples of humectants that can be used with the compositions of the present invention include amino acids, chondroitin sulfate, diglycerin, erythritol, fructose, glucose, glycerol polymers, ethylene glycol, 1,2, 6-hexanetriol, honey, hyaluronic acid, hydrogenated honey, hydrogenated starch hydrolysate, inositol, lactitol, maltitol, maltose, mannitol, natural moisturizing factor, PEG-15 butanediol, polyglycerol sorbitol, saccharide isomerate, salts of pyrrolidone carboxylic acid, potassium PCA, propylene glycol, isomerose, sodium glucuronate, sodium PCA, sorbitol, sucrose, trehalose, urea, and xylitol.

Other examples include acetylated lanolin, acetylated lanolin alcohol, alanine, algae extract, Aloe vera (Aloe barbadensis), Aloe vera extract, Aloe vera gel, Althea officinalis extract (Aloe officinalis), apricot (Prunus armeniaca) kernel oil, arginine aspartate, Arnica montana (Arnica montana) extract, aspartic acid, avocado (avocado, Persea gratissima) oil, barrier sphingolipid (barrier sphingolipid), butanol, beeswax, behenyl alcohol, beta-sitosterol, birch (Betula aluta, Betula alba) bark extract, borage (Borago officinalis) extract, pseudophylla (ruus aculeatus) extract, butylene glycol, Calendula (Calendula officinalis) extract, Calendula officinalis oil, candelilla (berberia) wax, caprylic acid/capric acid wax, canola oil (myristica fragrans) wax, canola oil, capric acid triglyceride, canola oil, capric acid, canola oil, capric acid triglyceride, canola oil, castor oil, safflower oil, castor oil, Carrot (Daucus carota sativa) oil, castor (Ricinus communis) oil, ceramide, ozokerite, ceteareth-5, ceteareth-12, ceteareth-20, cetearyl octanoate, ceteareth-20, ceteareth-24, cetoacetate, cetearyl octanoate, cetearyl palmitate, chamomile (Anthemis nobilis) oil, cholesterol ester, cholesteryl hydroxystearate, citric acid, sage (Salvia officinalis) oil, cocoa (Thermoroma cacao) fat, coconut oil-octanoate/caprate, coconut oil (Cocos nucifera) oil, collagen amino acids, corn (Zea mays) oil, fatty acids, decanoate, dimethicone copolyol, dimethicone, dioctyl adipate, Dioctyl succinate, dipentaerythritol hexacaprylate/hexacaprylate, DNA, erythritol, ethoxydiglycol, ethyl linoleate, Eucalyptus globulus oil, evening primrose oil, fatty acids, Geranium maculatum oil, glucosamine, glucoglutamate, glutamic acid, glyceryl polyether-26, glycerol distearate, glycerol hydroxystearate, glycerol laurate, glycerol linoleate, glycerol myristate, glycerol oleate, glycerol stearate SE, glycine, glycol stearate SE, glycosaminoglycans, grape (vitviferia) seed oil, hazelnut (Corylus americanus ) nut oil, hazelnut (Corylus avellana) nut oil, hyaluronic acid, hexylene glycol, hyaluronic acid, safflower (Carthamus sativus oil) nut oil, erythritol, ethoxydiglycol, ethyl linoleate, Eucalyptus globulus oil, evening primrose oil, fatty acids, Geranium grandis (Geranium grandis) oil, Geranium grandis oil, Geranium grandis, Hydrogenated castor oil, hydrogenated coconut oil glyceride, hydrogenated coconut oil, hydrogenated lanolin, hydrogenated lecithin, hydrogenated palm oil glyceride, hydrogenated palm kernel oil, hydrogenated soybean oil, hydrogenated tallow glyceride, hydrogenated vegetable oil, hydrolyzed collagen, hydrolyzed elastin, hydrolyzed glycosaminoglycans, hydrolyzed keratin, hydrolyzed soy protein, hydroxylated lanolin, hydroxyproline, isocetyl stearate, isocetyl stearoyloxystearate, isodecyl oleate, isopropyl isostearate, isopropyl lanolate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, isostearamide DEA, isostearic acid, isostearyl lactate, isostearyl pivalate, jasmine (Jasminum officinale ) oil, jojoba (Buxus chinensis) oil, kelp, shizandra (Aleurites moluccana) nut oil, lactamide MEA, hydrogenated palm oil, hydrogenated palm kernel oil, hydrogenated soybean oil, hydrogenated palm kernel oil, hydrogenated soybean oil, hydrogenated tallow glyceride, hydrogenated vegetable oil, hydrolyzed collagen, hydrolyzed elastin, hydrolyzed glycosaminoglycans, hydrolyzed keratin, hydrolyzed soy protein, hydroxylated lanolin, isopropyl oleate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, isopropyl oleate, isopropyl stearate DEA, isostearyl lactate, stearyl alcohol pivalate, jasmonate, Jasminum grandis (Jasminum, Jasminum officinale) oil, and mixtures thereof, Lanolin alcohol polyether-16, lanolin alcohol polyether-10 acetate, lanolin acid, lanolin alcohol, lanolin oil, lanolin wax, lavender (lavandala angustifolia) oil, lecithin, lemon (Citrus medica limonum) oil, linoleic acid, linolenic acid, Macadamia nut (Macadamia ternifolia) oil, maltitol, chamomile (chamomila recutita) oil, methylgluconate, methyl PCA ester, mineral oil, mink oil, Mortierella tawnensis oil, myristyl lactate, myristyl myristate, myristyl propionate, neopentyl glycol dicaprylate/dicaprate, octyldodecanol myristate, octyldodecanol stearate, octyl hydroxystearate, octyl palmitate, octyl salicylate, octyl stearate, oleic acid, olive (Olea europae) oil, orange (Citrus aurantium) oil, Palm (Elaeis guineensis) oil, palmitic acid, pantethine, panthenol, panthenyl ethyl ether, paraffin, PCA, peach (Prunus persica) kernel oil, peanut (Arachis Hypogaea) oil, PEG-8C12-18 acid ester, PEG-15 cocoamine, PEG-150 distearate, PEG-60 glyceryl isostearate, PEG-5 glyceryl stearate, PEG-30 glyceryl stearate, PEG-7 hydrogenated castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-20 methyl glucose sesquistearate, PEG-40 sorbitan monooleate, PEG-5 soybean, PEG-10 soybean sterol, PEG-2 stearate, PEG-8 stearate, PEG-20 stearate, PEG-32 stearate, PEG-40 stearate, PEG-50 stearate, panthenyl ethyl ether, hydrogenated castor oil, hydrogenated oil, castor oil, hydrogenated castor oil, PEG-20 methyl glucose sesquioleate, PEG-40 sorbitan monooleate, castor oil, castor, PEG-100 stearate, PEG-150 stearate, pentadecanolide, peppermint (Mentha piperita) oil, petrolatum, phospholipids, plankton extracts, polyamino acid polysaccharide condensates, polyglyceryl-3 diisostearate, polyquaternium-24, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 85, potassium myristate, potassium palmitate, propylene glycol dicaprylate/dicaprate, propylene glycol dicaprylate, propylene glycol dinonanoate, propylene glycol laurate, propylene glycol stearate SE, PVP, pyridoxine dipalmitate, retinol palmitate, rice (Oryza sativa) bran oil, RNA, rosemary (Rosmarinus officinalis) oil, rose oil, safflower (Carthamus tinctorius) oil, sage (Salvia officinalis) oil, and the like, Sandalwood (Santalum album) oil, serine, serum albumin, sesame (Sesamum indicum) oil, Butyrospermum parkii (Butyrospermum parkii) fat, silk powder, sodium chondroitin sulfate, sodium hyaluronate, sodium lactate, sodium palmitate, sodium PCA, sodium polyglutamate, soluble collagen, sorbitan laurate, sorbitan oleate, sorbitan palmitate, sorbitan sesquioleate, sorbitan stearate, sorbitol, soybean (Glycine soja) oil, sphingolipids, squalane, squalene, stearamide MEA-stearate, stearic acid, stearyloxypolydimethylsiloxane, stearyloxytrimethylsilane, stearyl alcohol, stearyl glycyrrhetinate, stearyl alcohol heptanoate, stearyl alcohol stearate, sunflower (heliothus annuus) seed oil, sweet almond (Prunus amygdalus duis) oil, synthetic beeswax, tocopherol acetate, tocopherol linoleate, tocopherol oleate, tocopherol acetate, tocopherol acid, Tribehenate, tridecanol pivalate, tridecanol stearate, triethanolamine, tristearyl alcohol, urea, vegetable oil, water, wax, wheat (Triticum vulgare) germ oil, and ylang (Cananga odorata) oil.

c. Antioxidant agent

Non-limiting examples of antioxidants that can be used with the compositions of the present invention include acetylcysteine, ascorbyl polypeptide, ascorbyl dipalmitate, ascorbyl methylsilanol pectate, ascorbyl palmitate, ascorbyl stearate, BHA, BHT, t-butylhydroquinone, cysteine HCI, dipentylhydroquinone, di-t-butylhydroquinone, dicetylthiodipropionate, dioleyl tocopheryl methylsilanol, disodium ascorbyl sulfate, distearylthiodipropionate, ditridecylthiodipropionate, lauryl gallate, erythorbic acid, ascorbyl esters, ethyl ferulate, ferulic acid, gallic acid, hydroquinone, isooctyl thioglycolate, kojic acid, magnesium ascorbate, magnesium ascorbyl phosphate, methylsilanol ascorbate, natural plant antioxidants such as green tea or grape seed extract, vitamin C, vitamin D, vitamin C, vitamin A, vitamin C, vitamin A, Nordihydroguaiaretic acid, octyl gallate, benzylthioglycolic acid, potassium ascorbyl tocopheryl phosphate, potassium sulfite, propyl gallate, quinone, rosmarinic acid, sodium ascorbate, sodium bisulfite, sodium erythorbate, sodium metabisulfite, sodium sulfite, superoxide dismutase, sodium thioglycolate, sorbitolfural, thiodiglycol, thiodiglycide, thiodiacetic acid, thioglycolic acid, thiolactic acid, thiosalicylic acid, tocopheryl polyether-5, tocopheryl polyether-10, tocopheryl polyether-12, tocopheryl polyether-18, tocopheryl polyether-50, tocopherol, tocol tolterol, tocopheryl acetate, tocopheryl linoleate, tocopheryl nicotinate, tocopheryl succinate, and tris (nonylphenol) phosphite.

d. Structuring agent

In other non-limiting aspects, the compositions of the present invention may comprise a structuring agent. In certain aspects, the structuring agent helps provide rheological characteristics to the composition to promote stability of the composition. In other aspects, the structuring agent may also function as an emulsifier or surfactant. Non-limiting examples of structuring agents include stearic acid, palmitic acid, stearyl alcohol, cetyl alcohol, behenyl alcohol, stearic acid, palmitic acid, the polyethylene glycol ether of stearyl alcohol having an average of about 1 to about 21 ethylene oxide units, the polyethylene glycol ether of cetyl alcohol having an average of about 1 to about 5 ethylene oxide units, and mixtures thereof.

e. Emulsifier

In certain aspects of the invention, the composition does not comprise an emulsifier. However, in other aspects, the composition may comprise one or more than one emulsifier. The emulsifier can reduce interfacial tension between phases and improve the formulation and stability of the emulsion. The emulsifiers may be nonionic, cationic, anionic and zwitterionic emulsifiers (see McCutcheon's (1986); U.S. Pat. No. 5011681; 4421769; 3755560). Non-limiting examples include glycerol esters, propylene glycol esters, fatty acid esters of polyethylene glycol, fatty acid esters of polypropylene glycol, esters of sorbitol, sorbitan esters, carboxylic acid copolymers, esters and ethers of glucose, ethoxylated esters, ethoxylated alcohols, alkyl phosphates, polyoxyethylene fatty ether phosphate esters, fatty acid amides, acyl lactylates, fatty acid salts, TEA stearate, DEA salt of oleyl polyether-3 phosphate, DEA salt of polyethylene glycol 20 sorbitan monolaurate (polysorbate 20), polyethylene glycol 5 soya sterol, steareth-2, steareth-20, steareth-21, ceteth-20, cetearyl glucoside, cetearyl alcohol, C12-13, PPG-2 methyl glucose ether distearate, PPG 5 ceteth-20, bis-PEG/PPG-20/20 dimethicone, ceteth-10, polysorbate 80, cetyl phosphate, potassium cetyl phosphate, diethanolamine cetyl phosphate, polysorbate 60, glyceryl stearate, PEG-100 stearate, arachidyl alcohol glucoside, and mixtures thereof.

f. Silicone-containing compound

In a non-limiting aspect, the silicone-containing compound includes any member of a family of polymer products in which the molecular backbone is comprised of alternating silicon and oxygen atoms with pendant groups attached to the silicon atoms. By varying the length of the-Si-O-chains, the pendant groups, and the cross-linking, silicones can be synthesized as a wide variety of materials. Their consistency can vary from liquid to gel to solid.

Silicone-containing compounds that can be used in the context of the present invention include those described in the specification or known to one of ordinary skill in the art. Non-limiting examples include silicone oils (e.g., volatile and non-volatile oils), gels, and solids. In some aspects, the silicon-containing compound comprises a silicone oil, such as a polyorganosiloxane. Non-limiting examples of polyorganosiloxanes include polydimethylsiloxane, cyclomethicone, silicone-11, phenyl trimethicone, trimethylsilylaminomethylsiloxane, stearyloxytrimethylsilane, or mixtures thereof, and other organosiloxane materials in any given ratio to achieve a desired consistency and application characteristics depending on the intended application (e.g., application to a particular area such as skin, hair, or eyes). "volatile silicone oils" include silicone oils having a low heat of vaporization, i.e., typically less than about 50 calories per gram of silicone oil. Non-limiting examples of volatile silicone oils include: cyclopolydimethylsiloxanes such as Dow Corning 344Fluid, Dow Corning 345Fluid, Dow Corning 244Fluid and Dow Corning 245Fluid, vollate Silicon 7207 (Union Carbide Corp., Danbury, Connecticut); low viscosity polydimethylsiloxanes, i.e., polydimethylsiloxanes having a viscosity of about 50cst or less than about 50cst (e.g., polydimethylsiloxanes such as Dow Corning 200-0.5cst Fluid). Dow Corning Fluid is commercially available from Dow Corning Corporation of Midland, Mich. In the third edition of the CTFA cosmetic ingredient dictionary, incorporated by reference, cyclomethicones and polydimethylsiloxanes are described as mixtures of cyclic dimethylpolysiloxane compounds and fully methylated linear siloxane polymers end-capped with trimethylsiloxy units, respectively. Other non-limiting volatile Silicone oils that can be used in the context of the present invention include those available from General Electric co, silicon Products div, walford, new york, and SWS Silicones div, Stauffer Chemical co, edlien, michigan.

g. Exfoliating agent

Keratolytic agents contain ingredients that remove dead skin cells on the outer surface of the skin. These agents may act mechanically, chemically, and/or otherwise. Non-limiting examples of mechanical exfoliants include abrasives such as pumice, silica, textiles, paper, shells, beads, solid crystals, solid polymers, and the like. Non-limiting examples of chemical exfoliants include acid and enzymatic exfoliants. Acids useful as exfoliants include, but are not limited to, glycolic acid, lactic acid, citric acid, alpha hydroxy acids, beta hydroxy acids, and the like. Other keratolytic agents known to those skilled in the art are also contemplated for use in the context of the present invention.

h. Essential oil

Essential oils include oils from herbs, flowers, trees, and other plants. Such oils typically exist as tiny droplets between plant cells and can be extracted by several methods known to those skilled in the art (e.g., steam distillation, liposuction (i.e., by using fat extraction), maceration, solvent extraction, or mechanical pressing). These types of oils tend to volatilize (i.e., are volatile oils) when exposed to air. Thus, while many essential oils are colorless, they oxidize and become darker in color over time. Essential oils are insoluble in water, but soluble in alcohols, ethers, non-volatile oils (vegetable oils) and other organic solvents. Typical physical characteristics of essential oils include a boiling point of about 160 ℃ to 240 ℃ and a density of about 0.759 to about 1.096.

Essential oils are generally named by the plant in which they are found. For example, rose oil or peppermint oil is from rose or peppermint plants, respectively. Non-limiting examples of essential oils that may be used in the context of the present invention include sesame oil, macadamia nut oil, tea tree oil, evening primrose oil, spanish sage oil, spanish rosemary oil, coriander oil, thyme oil, allspice oil, rose oil, fennel oil, balsamic flower oil, bergamot oil, rosewood oil, cedar oil, chamomile oil, sage oil, clary sage oil, clove oil, cedar oil, eucalyptus oil, fennel oil, sea fennel oil, frankincense oil, geranium oil, ginger oil, grapefruit oil, jasmine oil, juniper oil, lavender oil, lemon oil, lemongrass oil, lime berry oil, mandarin oil, marjoram oil, myrrh oil, neroli oil, orange oil, patchouli oil, pepper oil, black pepper oil, bitter orange leaf oil, pine oil, rose oil, rosemary oil, sandalwood oil, spearmint oil, spikenard oil, vetiver oil, wintergreen oil, ylang-ylang oil. Other essential oils known to those skilled in the art are also contemplated to be useful in the context of the present invention.

i. Thickening agent

Thickeners, including thickeners or gelling agents, include substances that increase the viscosity of the composition. Thickeners include agents that can increase the viscosity of the composition without substantially altering the efficacy of the active ingredients within the composition. Thickeners may also increase the stability of the compositions of the present invention. In some aspects of the invention, the thickener comprises hydrogenated polyisobutylene, trihydroxystearin, ammonium acryloyldimethyl taurate/VP copolymer, or mixtures thereof.

Non-limiting examples of additional thickeners that may be used in the context of the present invention include carboxylic acid polymers, crosslinked polyacrylate polymers, polyacrylamide polymers, polysaccharides, and gums. Examples of carboxylic acid polymers include crosslinked compounds containing one or more than one monomer derived from acrylic acid, substituted acrylic acids, and salts and esters of these acrylic acids and substituted acrylic acids, wherein the crosslinking agent contains two or more carbon-carbon double bonds and is derived from a polyol (see U.S. Pat. No. 5087445; No. 4509949; No. 2798053; CTFA International cosmetic ingredient dictionary, fourth edition, 1991, pages 12 and 80). Examples of commercially available carboxylic acid polymers include carbomers, which are homopolymers of acrylic acid crosslinked with allyl ethers of sucrose or pentaerythritol (e.g., CARBOPOL available from b.f. goodrich^TM900 series).

Non-limiting examples of crosslinked polyacrylate polymers include cationic and nonionic polymers. Examples are described in U.S. patent nos. 5100660, 4849484, 4835206, 4628078, 4599379).

Non-limiting examples of polyacrylamide polymers (including nonionic polyacrylamide polymers, including substituted branched or unbranched polymers) include polyacrylamide, isoparaffin and lauryl polyoxyether-7, acrylamide, and multi-block copolymers of substituted acrylamide with acrylic acid and substituted acrylic acid.

Non-limiting examples of polysaccharides include cellulose, carboxymethyl hydroxyethyl cellulose, cellulose acetate propionate, hydroxyethyl cellulose, hydroxyethyl ethyl cellulose, hydroxypropyl methyl cellulose, methyl hydroxyethyl cellulose, microcrystalline cellulose, sodium cellulose sulfate, and mixtures thereof. Other examples are alkyl substituted celluloses, wherein the hydroxyl groups of the cellulose polymer are hydroxyalkylated (preferably hydroxyethylated or hydroxypropylated) to form a hydroxyalkylated cellulose, which is subsequently treated with C₁₀-C₃₀The linear or branched alkyl groups are further modified by ether linkages. Typically these polymers are C₁₀-C₃₀Ethers of straight-chain or branched alcohols with hydroxyalkyl celluloses. Other useful polysaccharides include scleroglucans, which comprise a linear chain of (1-3) linked glucose units, with one (1-6) linked glucose per three units.

Non-limiting examples of gums that may be used in the present invention include gum arabic, agar, algin, alginic acid, ammonium alginate, pullulan, calcium alginate, calcium carrageenan, carnitine, carrageenan, dextrin, gelatin, gellan gum, guar hydroxypropyltrimonium chloride, hectorite, hyaluronic acid, hydrated silica, hydroxypropyl chitosan, hydroxypropyl guar, caraya gum, macroalgae, locust bean gum, natto gum, potassium alginate, potassium carrageenan, propylene glycol alginate, sclerotium gum, sodium carboxymethyl dextran, sodium carrageenan, tragacanth gum, xanthan gum, and mixtures thereof.

j. Preservative

Non-limiting examples of preservatives that can be used in the context of the present invention include quaternary ammonium preservatives, such as polyquaternium-1 and benzalkonium halides (e.g., benzalkonium chloride ("BAC") and benzalkonium bromide), parabens (e.g., methyl and propyl parabens), phenoxyethanol, benzyl alcohol, chlorobutanol, phenol, sorbic acid, thimerosal, or combinations thereof.

2. Pharmaceutical composition

It is also contemplated that pharmaceutically active agents may be used in the compositions of the present invention. Non-limiting examples of pharmaceutically active agents include anti-acne agents, agents for treating rosacea, analgesics, anesthetics, anorectics, antihistamines, anti-inflammatory agents including non-steroidal anti-inflammatory agents, antibiotics, antifungal agents, antivirals, antimicrobial agents, anticancer agents, anti-sarcoptidosis agents, pediculicides, antineoplastics, antiperspirants, antipruritics, antipsoriatic agents, anti-seborrheics, biologically active proteins and polypeptides, burn treatment agents, cauterizing agents, depigmenting agents, depilatory agents, diaper rash treatment agents, enzymes, hair growth stimulants, hair growth inhibitors including DFMO and its salts and analogs, hemostats, keratolytics, aphtha treatment agents, cold sore treatment agents, dental and periodontal treatment agents, light sensitive actives, skin protectants/barriers, steroids including hormones and corticosteroids, Sunburn treating agent, sunscreen agent, transdermal active agent, nasal active agent, vaginal active agent, wart treating agent, wound healing agent, etc.

F. Reagent kit

Kits are also contemplated for use in particular aspects of the invention. For example, the compositions of the present invention may be contained within a kit. The kit may comprise a container. The container may comprise a bottle, metal tube, laminated tube, plastic tube, dispenser, high pressure container, barrier container, bag, compartment, lipstick container, compact container, cosmetic tray capable of holding a cosmetic composition or other type of container such as an injection or blow molded plastic container in which the dispersion or composition is held or a desired bottle, dispenser or package/bag. The kit and/or container may comprise indicia on its surface. For example, the indicia may be words, phrases, abbreviations, pictures or symbols.

The container may dispense a predetermined amount of the composition. In other embodiments, the container (e.g., metal, laminated or plastic tube) may be squeezed to dispense the desired amount of the composition. The composition may be dispensed as a spray, aerosol, liquid, fluid or semi-solid form. The container may have a spraying mechanism, a pump mechanism or a squeezing mechanism. The kit can also include instructions for using the kit components and using any of the compositions contained in the containers. The instructions may include instructions for how to apply, use, and preserve the composition.

Examples

The following examples are set forth to illustrate preferred embodiments of the present invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Example 1

(materials used)

The active ingredients in table 1 were used to obtain the in vitro data mentioned below.

TABLE 1

Example 2

(in vitro study)

It has been determined that the combination of date palm extract, tea tree essential oil, millettia extract, and saccharide isomerate can inhibit nitric oxide synthase, increase alpha 2A adrenergic receptor agonist activity, reduce oxidation, increase antioxidant capacity of the composition or skin, inhibit COX-2 production, inhibit VEGF production, inhibit IL-6 production, inhibit IL-8 production, reduce TNF-alpha production, increase collagen stimulation, increase lysyl oxidase expression, inhibit MMP1 activity, increase occludin production, increase filaggrin production, and/or increase skin moisturization. A summary of the results is shown in table 2, and the methods for determining the properties of the ingredients are provided below.

TABLE 2

Alpha 2A adrenergic receptor agonist assay: date extract was shown to increase alpha 2A adrenergic receptor agonist activity. Adrenergic receptors are metabotropic (i.e., metabolism affecting) G protein-coupled receptors (GPCRs). Many types of cells have such receptors, and binding of agonists (activators) often leads to sympathetic responses, depending on the affected receptor (e.g., heart rate modification, vasoconstriction/dilation, changes in fat metabolism, etc.). Agonism of the α 2A adrenergic receptors on smooth muscle cells can result in a vasoconstrictive effect-i.e., the blood vessels in which these cells and receptors are located will constrict and limit the pooling of blood and lymph in this area, which can result in an improved and uniform appearance of the skin in this area and reduced dark under-eye circles and symptoms of rosacea.

Cell Dielectric Spectroscopy (CDS) is a technique that measures the change in impedance caused by activation of target cell receptors in cell culture. The CDS was used to determine the activity of α 2A adrenergic receptors in recombinant Chinese Hamster Ovary (CHO) cells expressing the human α 2A receptor. Changes induced by treatment with the embodiments were compared to the effect of a positive control reference activator (100nM epinephrine) and expressed as a percentage of the positive control reference effect ((measured specific response/control specific agonist response) × 100). Date palm extract was shown to enhance the activity of alpha 2A adrenergic receptor agonists, potentially affecting capillary vasoconstriction and alleviating dark under-eye circles and rosacea. Specifically, 0.5% of the date extract increased the α 2A adrenergic receptor activity of the positive control reaction by 24%, 1.5% of the date extract increased the α 2A adrenergic receptor activity of the positive control reaction by 65%, and 4.5% of the date extract increased the α 2A adrenergic receptor activity of the positive control reaction by 102%. See table 2.

Nitric oxide synthase activity assay: the saccharide isomerate and the extract of Artocarpus heterophyllus show a reduction in nitric oxide synthase activity. Nitric Oxide (NO) is a reactive free radical that plays an important role in many key physiological functions. NO is involved in host defense and development, activation of regulatory proteins, and direct covalent interactions with functional biomolecules. Nitric Oxide Synthase (NOS) is an important mediator of vasodilation in blood vessels. NOS is a family of enzymes that catalyze the production of Nitric Oxide (NO) from L-arginine. The NOS assay (Abnova, KA1634) comprises two steps: the NOS reaction step, during which NO is produced, is followed by the NO detection step. Since NO produced by NOS is rapidly oxidized to nitrite and nitrate, NO production was measured after reducing nitrate to nitrite using a modified Griess method. The effect of saccharide isomers and extracts of Artocarpus heterophyllus on NO production by NOS was analyzed by bioassay. Saccharide isomerate and extracts of Artocarpus heterophyllus were shown to reduce NOS activity more than 1 μ M SIN-1 activity control. Specifically, the saccharide isomerate reduced NOS activity by about 45%, the extract of mataire reduced NOS activity by about 36%, while the positive control reduced NOS activity by only about 29% (not shown). See table 2.

Tumor necrosis factor alpha (TNF- α) assay: date extract, saccharide isomerate and millettia extract were shown to inhibit the production of TNF-alpha. The prototype ligand of the TNF superfamily, TNF- α, is a pleiotropic cytokine that plays a major role in inflammation. The increase in its expression is associated with an upregulation of pro-inflammatory activity. The bioassay is used for analyzing the effect of date extract, saccharide isomerate and Artocarpus heterophyllus extract on the production of TNF-alpha by human epidermal keratinocytes. The endpoint of this assay is a spectrophotometric measurement reflecting the presence of TNF-alpha and cell viability. The determination adopts a quantitative sandwich enzyme immunoassay technology, and the TNF-alpha specific monoclonal antibody is coated on a micropore plate in advance. Standards and samples were pipetted into the wells and all TNF-alpha present was bound by the immobilized antibody. After washing away all unbound material, TNF-alpha specific enzyme-linked polyclonal antibodies were added to the wells. After washing to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and the color is developed as a proportion of the amount of TNF-. alpha.bound in the initial step. The color development was stopped, and the color development intensity was measured. Detection was performed with a microplate reader at 450 nm.

5% CO at 37 ℃₂InSub-confluent normal human adult keratinocytes (Cascade Biologics) cultured in standard growth medium (Cascade Biologics) were treated with phorbol-12-myristate-13-acetate (PMA, 10ng/ml, Sigma Chemical, # P1585-1MG) in the presence or absence of test extracts or controls for 6 hours. PMA resulted in a significant increase in TNF- α secretion, peaking at 6 hours post-treatment. After incubation, the cell culture medium was collected and used as described above from R&Sandwich enzyme-linked immunosorbent assay (ELISA) of D Systems (# DTA00C) quantitated the secreted amount of TNF- α. The results show that date extract, saccharide isomerate and millettia extract reduced the production of TNF-alpha. Specifically, the date extract reduced the production of TNF- α by about 84.6%, the saccharide isomerate reduced the production of TNF- α by about 88%, and the millettia extract reduced the production of TNF- α by about 80%. See table 2.

Production of occludin-1: saccharide isomerate showed increased cutinThe production of occludin-1 in the forming cells. Occludin-1 is a protein important for the formation of tight junctions and the skin's moisture barrier function. The production of occludin-1 in treated and untreated keratinocytes was determined using a bioassay to analyze the concentration of occludin-1 in the keratinocyte lysates. Use ofSIMON^TMThe bioassay was performed by western blotting. For the sample, at 37 ℃ and 5% CO₂Next, adult human epidermal keratinocytes (HEKa) derived from Life Technologies (C-005-5C) were treated in a medium containing calcium derived from Life Technologies (M-EP-500-CA) supplemented with keratinocyte growth supplement (HKGS) derived from Life Technologies (S-101-5)Growth medium was grown for 24 hours. HEKa was then added to a negative control containing test compound, no compound, or 1mM CaCl₂The positive control of (4) is incubated in the growth medium for 24 hours to 48 hours. The NEKa cells were then washed, collected and stored in ice or colder environment until lysed on ice using lysis buffer and sonication. The protein concentration of the samples was determined and used to normalize the samples. The cell lysate was stored at-80 ℃ until used in the bioassay.

SIMON^TMWestern blot bioanalysis quantitative western blot analysis techniques using antibodies specific for occludin to quantitatively detect occludin in a sample were used. Cell samples were lysed and protein concentrations were normalized as described above. The normalized sample and molecular weight standards were loaded and run on a denatured protein separation gel using capillary electrophoresis. The proteins in the gel are then immobilized and immunodetected using a first antibody specific for occludin. Then, an enzyme that binds to the primary antibody is usedThe co-detection antibody immunodetects the immobilized protein. The chemiluminescent substrate solution is then added to the immobilized protein to allow for the development of chemiluminescence in proportion to the amount of occludin bound in the immobilization. Chemiluminescence development was stopped at a specified time, and the intensity of the chemiluminescence signal was measured and compared to positive and negative controls. The results indicate that the saccharide isomerate increased production of occludin-1 by 100%. See table 2.

Production of filaggrin: saccharide isomers have been shown to increase production of filaggrin. Filaggrin is a precursor of Natural Moisturizing Factor (NMF) in the skin. Increasing NMF increases the moisture content of the skin. The silk fibroin concentration in keratinocyte lysates was analyzed using bioassay to determine silk fibroin production in treated and untreated keratinocytes. Use ofSimon^TMWestern blotting was used to quantify the production of filaggrin. For each sample, Normal Human Epidermal Keratinocytes (NHEK) were plated on calcium-containing EPI-200-Mattek from Life Technologies (M-EP-500-CA)Growing in a growth medium. 5% CO at 37 ℃ before treatment₂NHEK cells were incubated overnight in growth medium. The NHEK cells were then incubated in growth medium with 1% test compound/extract or growth medium without compound/extract (negative control) for 24 to 36 hours. The NHEK cells were then washed, collected and stored in ice or colder environment until lysed on ice using lysis buffer nuclear sonication. The protein concentration of the samples was determined and used to normalize the samples. The lysate can be stored at-80 ℃ until use in a quantitative assay.

Simon^TMProteinThe western blot bioanalysis uses quantitative western blot immunoassay techniques that use antibodies specific for silk fibroin to quantitatively detect silk fibroin in a sample. Cell samples were lysed and protein concentrations were normalized. The normalized sample and molecular weight standards were loaded and run on a denatured protein separation gel using capillary electrophoresis. The proteins in the gel are immobilized and immunologically detected using a first antibody specific for filaggrin. The immobilized protein is then immunodetected with an enzyme-linked detection antibody that binds the first antibody. The chemiluminescent substrate solution is then added to the immobilized protein to allow for the development of chemiluminescence in proportion to the amount of filaggrin bound in the immobilization. Chemiluminescence development was stopped at a specified time, and the intensity of the chemiluminescence signal was measured and compared to positive and negative controls. The results show that the saccharide isomer increased the production of filaggrin by 27%. See table 2.

Moisture/hydration measurements: using the skin moisture/hydration assay, saccharide isomers showed an increase in the skin moisture measurement. The assay uses a Nova derm Phase Meter for impedance measurements. Moisture content changes of three-dimensional cultured artificial skin equivalents of tissue engineering (MatTek Corporation) were measured with an impedance meter. The outer layers of skin have different electrical properties. When the skin is dry, it is poorly conductive. As it becomes more moist, the conductivity increases. Thus, changes in impedance (related to conductivity) are used to evaluate changes in hydration of the artificial skin equivalent.

For this assay, treated and untreated artificial skin equivalents were used. The Nova derm Phase Meter was calibrated on each test day according to the instrument instructions. For comparison purposes, temperature and relative humidity were labeled. Impedance evaluation was performed as follows: prior to measurement, the samples are equilibrated in a chamber with a defined humidity (e.g. 30% to 50%) and a defined temperature (e.g. 68 ℃ to 72 ℃). Impedance readings were taken for each sample, recorded and averaged. The impedance meter was set using T5, which averaged the impedance value every five seconds after application to the sample. The reported changes were statistically different and significant. Saccharide isomerate was measured to increase the conductance by 80%, indicating an increase in moisture/hydration. See table 2.

Antioxidant (AO) assay: date extract was shown to reduce oxidation of at least one biomarker indicative of total antioxidant capacity. The antioxidant system of living organisms includes enzymes such as superoxide dismutase, catalase, and glutathione peroxidase; macromolecules such as albumin, ceruloplasmin, and ferritin; and a range of small molecules including ascorbic acid, alpha-tocopherol, beta-carotene, reduced glutathione, uric acid, and bilirubin. The total amount of endogenous and food-derived antioxidants represents the total antioxidant capacity of the extracellular fluid. The cooperation of all the different antioxidants provides greater protection against attack by reactive oxygen or nitrogen radicals than any of the individual compounds alone. Thus, the total antioxidant capacity may give more relevant biological information than the antioxidant capacity obtained by measuring the individual components, since it takes into account the cumulative effect of all antioxidants present in plasma and body fluids. Antioxidant capacity kit #709001 from Cayman Chemical (anaba, michigan, USA) was used in an in vitro bioassay to measure the total antioxidant capacity of date palm extract. The assay relies on antioxidant inhibition in the sample(2,2' -biazo-bis- [ 3-ethylbenzothiazoline sulfonate salt]) Is oxidized by the positive iron myoglobin+ to the capacity of the cell. Antioxidant prevention in test samplesThe capacity for oxidation was compared to the water soluble tocopherol analogue Trolox and quantified as molar equivalents of Trolox. The protocol may be performed according to manufacturer recommendations. The oxidation potential of date palm extract was determined to be 99.25% compared to Trolox. See table 2.

Cyclooxygenase (COX) assay: date extract was shown to inhibit COX-2 production. COX (ovine) colorimetric inhibitor screening assay (#760111, Cayman Chemical) was used to determine inhibition of cyclooxygenase-1 and cyclooxygenase-2 (COX-1, COX-2). COX is a bifunctional enzyme with cyclooxygenase activity and peroxidase activity. Cyclooxygenase activity converts arachidonic acid to hydroperoxides endoperoxide (prostaglandin G2; PGG2), and the peroxidase component reduces endoperoxide (prostaglandin H2; PGH2) to the corresponding alcohol, prostaglandin precursor, thromboxane and prostacyclin. The COX inhibitor screening assay measures the peroxidase component of cyclooxygenase. Peroxidase activity was determined by colorimetrically monitoring the appearance of oxidized N, N' -tetramethyl-p-phenylenediamine (TMPD). The inhibitor screening assay contains both COX-1 and COX-2 enzymes to screen for isoenzyme specific inhibitors. Purified enzyme, heme and test extract were mixed in content detection buffer according to the manufacturer's instructions and incubated for 15 minutes at room temperature with shaking. After incubation, arachidonic acid and colorimetric substrate were added to start the reaction. The color change was assessed by reading the plates at 590 nm. The percent inhibition of COX-1 or COX-2 activity was calculated and compared to untreated controls to determine the ability of the date extract to inhibit the activity of the purified enzyme. The activity of date palm extract on COX-2 peroxidase was determined to be 85.02% inhibition. See table 2.

Cytokine arrays (including VEGF, IL-6 and IL-8): date extract has been shown to inhibit the production of IL-6, IL-8 and VEGF. IL-6 and IL-8 are cytokines involved in inflammatory and anti-inflammatory responses. IL-6 is an interleukin that functions as a proinflammatory cytokine and an anti-inflammatory cytokine. IL-8 is a key mediator associated with inflammation and acts as a chemotactic cytokine and inflammatory cytokine. VEGF is a cytokine that stimulates angiogenesis and revascularization, possibly contributing to inflammation, redness and rosacea. Inhibition of VEGF, IL-6 and IL-8 production was determined using protein detection assay using biotinylated antibodies to various cytokines for antibody detection. Date extract was determined to inhibit VEGF production by 86.4%, IL-6 expression by 73.4%, and IL-8 production by 97.15%. See table 2.

Briefly, human epidermal keratinocytes were cultured to 70% to 80% confluence. The medium in the plate was aspirated and 0.025% trypsin/EDTA was added. When the cells round, the dish was tapped to release the cells. Cells containing trypsin/EDTA were removed from the culture dish and equilibrated. Cells were centrifuged at 180 Xg for 5 min. The cells formed a pellet and the supernatant was aspirated. The obtained agglomerates areResuspended in culture medium (Cascade Biologics). Cells were seeded into 6-well plates at approximately 10% to 20% confluence. After the cells became approximately 80% confluent, the medium was aspirated and 1.0ml was addedAnd phorbol 13-tetradecanoic acid 12-acetate ("PMA") (a known inducer of inflammation) and a dilution of date extract were added to two duplicate wells. Diluting the test composition to a final volume of 1ml (Growth medium). The medium was gently swirled to ensure adequate mixing. In addition, 1.0ml ofTo control wells with or without additional PMA. After formulation, plates were then plated at 37. + -. 1 ℃ and 5.0. + -. 1% CO₂The incubation was continued for about 5 hours. After 5 hours incubation, the entire medium was collected into conical tubes and frozen at-70 ℃, the frozen medium was subsequently loaded onto dry ice.

On the day of analysis, 16-pad hybridization chambers were attached to 16-pad FAST slides in triplicate with 16 anti-cytokine antibodies (including VEGF, IL-6, and IL-8) and experimental controls (Whatman BioSciences), and the slides were placed in FAST^TMFrame (4 slides per Frame) for treatment. Using 70ml S at room temperature&Protein S assay blocking buffer (Whatman Schleicher and Scheull) blocks the array for 15 minutes.The blocking buffer was removed and 70ml of each supernatant sample was added to each array. The array was incubated at room temperature for 3 hours with gentle agitation. The array was washed 3 times with TBS-T. The arrays were treated with 70ml of an antibody mixture containing biotinylated antibodies corresponding to the capture antibodies of each array. The array was incubated at room temperature for 1 hour with gentle agitation. The array was washed 3 times with Tris buffer (TBS-T). The array was incubated with 70ml of a solution containing streptavidin-Cy 5 conjugate for 1 hour at room temperature with gentle agitation. The array was washed 3 times with TBS-T, rinsed rapidly in deionized water, and dried.

Slides in a Perkin-ElmerAnd (4) imaging in a 4000 confocal fluorescence imaging system. The array image was saved and analyzed with Imaging Research ArrayVision software. Briefly, spot intensities were determined by subtracting background signals. Duplicate spots from each sample condition were averaged and then aligned with the appropriate control.

Collagen stimulation assay: date extract showed increased collagen stimulation. Collagen is an extracellular matrix protein that is critical to skin structure. Increased collagen synthesis helps to improve skin firmness and elasticity. The bioassay method is used for examining the effect of the Phoenix dactylifera extract on the production of procollagen peptide (a collagen precursor) by human epidermal fibroblasts. The endpoint of this assay is a spectrophotometric measurement reflecting the presence of procollagen peptides and cell viability. The determination adopts a quantitative sandwich enzyme-linked immunoassay technology, and the procollagen peptide specific monoclonal antibody is coated on a micropore plate in advance. Standards and samples are pipetted into the wells and any procollagen peptide present is bound by the immobilized antibody. After washing away all unbound material, a procollagen peptide-specific enzyme-linked polyclonal antibody is added to the wells. After washing to remove all unbound antibody-enzyme reagent, a substrate solution is added to the wells to develop color in proportion to the amount of procollagen peptide bound in the initial step. Color development was stopped and the intensity of the color at 450nm was measured using a microplate reader.

To generate samples and controls, 10% CO at 37 ℃ in standard DMEM growth medium containing 10% fetal bovine serum (Mediatech)₂Culturing sub-confluent normal adult epidermal fibroblasts (Cascade Biologics). Cells were treated with date extract and control for 3 days. Following incubation, the cell culture medium was collected and the amount of secreted procollagen peptide was quantified using a sandwich enzyme-linked immunosorbent assay (ELISA) from Takara (# MK101) as described above. Date extract was determined to increase collagen stimulation by 25.3%. See table 2.

And (3) determining the content of lysyl oxidase: date extract showed increased expression of lysyl oxidase. Lysyl oxidase assays are performed on skin cells (e.g., epidermal keratinocytes, fibroblasts, and/or dermal endothelial cells) to determine the ability of date palm extract to stimulate the expression of lysyl oxidase in skin. Lysyl oxidase can catalyze the cross-linking of elastin and collagen, thereby providing a structurally stronger matrix for the skin. By increasing the expression of lysyl oxidase, increased crosslinking of elastin and collagen may occur, which may be beneficial in reducing the appearance of fine lines, wrinkles, skin laxity, and/or inelastic skin. Date palm extract was determined to increase lysyl oxidase expression by 38.37%. See table 2.

Matrix metalloproteinase 1 activity (MMP1) assay: date extract was shown to inhibit MMP1 activity. MMPs are extracellular proteases that play a role in many normal and disease states due to their broad substrate specificity. MMP1 substrates include collagen IV.

Invitrogen from Invitrogen^TMMolecular Probes^TMEnzChek^TMThe gelatinase/collagenase assay kit (# E12055) was designed as an in vitro assay to measure MMP1 enzyme activity. The kit utilizes a fluorescent gelatin substrate to detect MMP1 protease activity. This assay relies on the ability of the purified MMP1 enzyme to degrade fluorescent gelatin substrates. Once the substrate is specifically cleaved by MMP1, a bright green fluorescence is displayed and fluorescence is monitored using a fluorescent microplate reader. In the presence or absence of purified enzyme and substrateThe date extract was incubated under the conditions described above to determine its protease inhibitory activity. Date extract was determined to inhibit MMP1 activity by 70.34%. See table 2.

Example 3

(general preparation)

The combination of active ingredients may be included in a variety of topical product formulations for use on skin and/or hair. Tables 3 and 4 describe the general formulations or skin test formulations in which the active ingredients can be incorporated. These formulations can also be used to determine the type of benefit of the active ingredient to the skin. These formulations are prepared in a manner that ensures that all of the skin benefits resulting from topical application of the formulations to the skin can be directly attributed to the active ingredients tested. In the context of aspects of the present invention, the active ingredients that may be tested may be date palm extract, tea tree essential oil, millettia extract and saccharide isomers or any combination thereof, or all of these active ingredients, or at least 1, at least 2 at least, 3, at least 4 and/or at least 4 of said active ingredients. It will be appreciated that other standard test vehicles may also be used to determine the skin benefit properties of the active ingredient, and that the following formulations are non-limiting test vehicles.

TABLE 3 ANG

Composition (I)	% concentration (by weight)
		Phase A
Water (W)	84.80
		Xanthan gum	0.1
M-p-hydroxybenzoic acid ester	0.15
		P-P-hydroxybenzoic acid ester	0.1
Citric acid	0.1
		Phase B
Cetyl alcohol	4.0
		Glyceryl stearate + PEG 100	4.0
Palmitic acid octyl ester	4.0
		Polydimethylsiloxane	1.0
Tocopherol acetate	0.2
		Phase C
Active ingredient	2.0
		Total of	100

Procedure for preparation of the composition: the xanthan gum was sprinkled into water and mixed for 10 minutes. Then, add all ingredients in phase a and heat to 70 ℃ to 75 ℃. Add all the materials in phase B to a separate beaker and heat to 70 ℃ to 75 ℃. Phases A and B were mixed at 70 ℃ to 75 ℃. Stirring was continued and the composition was allowed to cool to 30 ℃. Then, add phase C ingredients with stirring.

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

TABLE 4 ANG

Add the ingredients in phase a to the beaker and heat to 70 ℃ to 75 ℃ with stirring. Then, add the ingredients of phase B with phase a and cool to 30 ℃ with stirring. Then, add phase C ingredients with stirring.

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

Example 4

(exemplary formulations)

The formulations shown in tables 5 to 10 are non-limiting examples of the types of formulations that may be prepared with the present invention. Such formulations may be prepared using any standard method. For example, simple mixing of the ingredients in a beaker may be used. The ingredients should be mixed and heated as necessary to obtain a homogeneous composition. The active ingredients that may be used in the formulation may include date extract, tea tree essential oil, millettia extract and saccharide isomers or any combination thereof, or all of these active ingredients, or at least 1, at least 2, at least 3, at least 4 and/or at least 4 of the active ingredients.

Table 5 includes non-limiting examples of compositions of the present invention. The compositions can be formulated as emulsions (e.g., o/w, w/o, o/w/o, w/o/w, etc.), and additional ingredients identified throughout this specification can be included in the compositions of table 5 (e.g., by adjusting the water content of the composition). In addition, the concentration ranges of the ingredients in table 5 may be varied depending on the desired formulation (e.g., cream, lotion, moisturizer, cleanser, etc.).

TABLE 5

Composition (I)	% concentration (by weight)
		Water (W)	Proper amount of
Active ingredient	0.1 to 10 percent
		Glycerol	3 to 40 percent
Butanediol	0.0001 to 10 percent
		Propylene glycol	0.0001 to 10 percent
Phenoxyethanol	0.0001 to 10 percent
		EDTA disodium salt	0.0001 to 10 percent
Steareth-20	0.0001 to 10 percent
		Chlorhexidine digluconate	0.0001 to 10 percent
Potassium sorbate	0.0001 to 10 percent
		Preservative	0.0001 to 2%
Total of	100

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

All preservatives identified in the present description or known in the art may be used.

Table 6 includes non-limiting examples of compositions of the present invention. The compositions can be formulated as emulsions (e.g., o/w, w/o, o/w/o, w/o/w, etc.), and additional ingredients identified throughout this specification can be included in the compositions of table 6 (e.g., by adjusting the water content of the composition). In addition, the concentration ranges of the ingredients identified in table 6 may vary depending on the desired formulation (e.g., cream, lotion, moisturizer, cleanser, etc.).

TABLE 6

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

All preservatives identified in the present description or known in the art may be used.

Table 7 includes non-limiting examples of compositions of the present invention. The compositions can be formulated as emulsions (e.g., o/w, w/o, o/w/o, w/o/w, etc.), and additional ingredients identified throughout this specification can be included in the compositions of table 7 (e.g., by adjusting the water content of the composition). In addition, the concentration ranges of the ingredients identified in table 7 may vary depending on the desired formulation (e.g., cream, lotion, moisturizer, cleanser, etc.). In particular embodiments, the composition of table 7 may be a moisturizer.

TABLE 7

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

Table 8 includes non-limiting examples of compositions of the present invention. The compositions can be formulated as emulsions (e.g., o/w, w/o, o/w/o, w/o/w, etc.), and additional ingredients identified throughout this specification can be included in the compositions of table 8 (e.g., by adjusting the water content of the composition). In addition, the concentration ranges of the ingredients identified in table 8 may vary depending on the desired formulation (e.g., cream, lotion, moisturizer, cleanser, etc.). In particular embodiments, the composition of table 8 may be a moisturizer.

TABLE 8

Composition (I)	% concentration (by weight)
		Water (W)	Proper amount of
Active ingredient	0.1 to 10 percent
		Glycerol	0.0001 to 10 percent
Pentanediol	0.0001 to 10 percent
		Octanediol	0.0001 to 10 percent
EDTA disodium salt	0.0001 to 10 percent
		Vaseline	0.0001 to 10 percent
Squalane	0.0001 to 10 percent
		Cetyl alcohol	0.0001 to 10 percent
ARLACELTM165	0.0001 to 10 percent
		Polydimethylsiloxane	0.0001 to 10 percent
SIMULGELTM600	0.0001 to 10 percent
		Total of	100

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

Table 9 includes non-limiting examples of compositions of the present invention. The compositions can be formulated as emulsions (e.g., o/w, w/o, o/w/o, w/o/w, etc.), and additional ingredients identified throughout this specification can be included in the compositions of table 9 (e.g., by adjusting the water content of the composition). In addition, the concentration ranges of the ingredients identified in table 9 may vary depending on the desired formulation (e.g., cream, lotion, moisturizer, cleanser, etc.). In particular embodiments, the composition of table 9 may be a sunscreen lotion.

TABLE 9

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

Sunscreen ingredients may be any sunscreen ingredient identified in this specification or a combination of such ingredients (e.g., UV absorbers and/or reflectors) or sunscreen ingredients known to one of ordinary skill in the art. In one embodiment, the sunscreen ingredient is a combination of zinc oxide and titanium dioxide.

Table 10 includes non-limiting examples of compositions of the present invention. Additional ingredients identified throughout this specification may be included in the compositions of table 10 (e.g., by adjusting the moisture content of the composition). In addition, the concentration ranges of the ingredients identified in table 10 may vary depending on the desired formulation (e.g., cream, lotion, moisturizer, cleanser, etc.). In particular embodiments, the composition of table 10 may be a detergent.

Watch 10

Composition (I)	% concentration (by weight)
		Water (W)	Proper amount of
Active ingredient	0.1 to 10 percent
		EDTA disodium salt	0.0001 to 10 percent
Citric acid	0.0001 to 10 percent
		Pentanediol	0.0001 to 10 percent
Octanediol	0.0001 to 10 percent
		Sodium methyl cocoyl taurate	10 to 30 percent
Cocoyl amphodiacetate sodium salt	1 to 10 percent
		Total of	100

The active ingredients identified throughout the specification may be incorporated into the composition as active ingredients. The active ingredients may be used alone or in combination in the composition. The concentration range of the active ingredient (or combination of active ingredients) can be varied as desired or needed by increasing or decreasing the amount of water.

Example 5

(additional measurement)

Assays that can be used to determine the efficacy of any one ingredient, or any combination of ingredients, or composition having a combination of ingredients disclosed throughout the specification and claims can be determined by methods known to those of ordinary skill in the art. The following are non-limiting assays that may be used in the context of the present invention. It should be appreciated that other testing procedures may be used, including, for example, objective and subjective procedures.

B16 pigmentation assay: melanin AProduction is the process by which melanocytes produce melanin, a naturally occurring pigment that imparts color to the skin, hair, and eyes. Inhibiting melanin production is beneficial for preventing skin darkening and reducing age-related dark spots. The bioassay used B16-F1 melanocytes (ATCC), an immortal mouse melanoma cell line, to analyze the effect of compounds on melanogenesis. The end point of this assay is the spectrophotometric measurement of melanin production and cellular activity. Can be 10% CO at 37 deg.C₂B16-F1 melanophores were incubated in standard DMEM growth medium containing 10% fetal bovine serum (Mediatech) and then treated with any one of the active ingredients, combinations of ingredients, or compositions having the combinations disclosed in this specification for 6 days. After incubation, melanin secretion was measured by absorption at 405nm and cellular activity was quantified.

Elastin stimulation assay: elastin is a connective tissue protein that helps the skin to recover shape after stretching or contraction. Elastin is also an important load-bearing protein for use in locations where storage of mechanical energy is required. Elastin is made by linking a number of soluble tropoelastin molecules in a lysyl oxidase-catalyzed reaction. Elastin secretion and elastin fibers were monitored in cultured human fibroblasts by staining the cultured human fibroblasts with immunofluorescent antibodies to elastin.

Laminin and fibronectin stimulation assays: laminin and fibronectin are the major proteins in the dermal-epidermal junction (DEJ), also known as the basement membrane band. The DEJ is located between the dermis and epidermis, and its junction forms a finger-like protrusion called the reticulum. Epidermal cells absorb their nutrients from the blood vessels of the dermis. The mesh ridges increase the surface area of the epidermis that is in contact with these blood vessels and the nutrients required. DEJ provides adhesion between two tissues and controls the structural integrity of the skin. Laminin and fibronectin are two structural glycoproteins located in the DEJ. Laminin and fibronectin are thought to be the adhesive that holds cells together, secreted by dermal fibroblasts, and help promote the intracellular and intercellular adhesion of epidermal cells to DEJ. The secretion of laminin and fibronectin can be monitored by quantifying laminin and fibronectin in the cell supernatant of cultured human fibroblasts treated for 3 days with or without 1.0% of the final concentration of the test component. After incubation, the laminin and fibronectin content can be measured in an enzyme-linked immunosorbent assay (ELISA) using an immunofluorescent antibody directed against each protein. Measurements of the metabolic activity of cells were normalized by the biotransformation of 3- (4, 5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazole (MTS).

ORAC determination: the oxidative radical absorption (or absorption) capacity (ORAC) of any of the active ingredients, combinations of ingredients, or compositions having the combinations disclosed herein can also be analyzed by measuring the antioxidant activity of the ingredients or compositions. Antioxidant activity indicates the ability to reduce the oxidizing agent (oxidizer). This assay quantifies the extent and length of time required to inhibit the action of, for example, oxidants such as oxygen radicals, which are known to cause damage to cells (e.g., skin cells). The ORAC value of any of the active ingredients, combinations of active ingredients, or compositions containing the combinations disclosed in this specification can be determined by methods known to those of ordinary skill in the art (see U.S. publication Nos. 2004/0109905 and 2005/0163880; and commercially available kits such as the Zen-Bio ORAC antioxidant assay kit (# AOX-2)). The Zen-Bio ORAC antioxidant assay kit measures the loss of fluorescein fluorescence over time due to the formation of peroxy radicals obtained by decomposition of AAPH (2,2' -azobis-2-methylpropionamidine dihydrochloride). A water-soluble vitamin E analog, Trolox, served as a positive control for inhibition of fluorescein decay in a dose-dependent manner.

Mushroom tyrosinase activity assay: in mammalian cells, tyrosinase catalyzes two steps in the multistep biosynthesis of melanin from tyrosine (and the polymerization of dopachrome). Tyrosinase is localized in melanoblasts and produces melanin (aromatic quinone compounds) that imparts color to the skin, hair, and eyes. Purified mushroom tyrosinase (Sigma) can be incubated with its substrate L-dopa (fisher) in the presence or absence of each of the active ingredients, combinations of any of the ingredients, or compositions with the combinations disclosed in this specification. Pigment formation can be assessed by colorimetric plate readings at 490 nm. The percent inhibition of mushroom tyrosinase activity was calculated and compared to untreated controls to determine the ability of the test component or combination thereof to inhibit purified enzyme activity. The inhibition of the test extracts was compared to that of kojic acid (Sigma).

Matrix metalloproteinase 3 and matrix metalloproteinase 9 enzyme activity (MMP 3; MMP9) assays: an in vitro Matrix Metalloproteinase (MMP) inhibition assay is disclosed. MMPs are extracellular proteases that function in many normal and disease states due to their broad substrate specificity. MMP3 substrates include collagen, fibronectin, and laminin; whereas MMP9 substrates include collagen VII, fibronectin, and laminin. Using a BioMol International colorimetric drug discovery kit for MMP3(AK-400) and MMP-9(AK-410), the assay was designed to measure the use of thiopeptides as chromogenic substrate (Ac-PLG- [ 2-mercapto-4-methylpentanoyl ™ 2)]LG-OC2H5)5, 6. The peptide bond of the MMP cleavage site is replaced by a thioester bond in a thiopeptide. Hydrolysis of this bond by MMP produces a sulfhydryl group which is reacted with DTNB [5,5 '-dithiobis (2-nitrobenzoic acid), Elmann's reagent]The reaction forms 2-nitro-5-thiobenzoic acid, which can be converted by reaction at 412nm (. epsilon.) < 13600M^-1cm^-1Absorbance at pH 6.0 and pH above 7). The active ingredients disclosed in the specification, any combination of the ingredients, or a composition having the combination can be assayed.

Lipoxygenase (LO) assay: an in vitro Lipoxygenase (LO) inhibition assay is disclosed. LO is an iron-containing dioxygenase, a non-heme enzyme, that catalyzes the addition of molecular oxygen to fatty acids. Linoleate and arachidonate are the main substrates of LO in plants and animals. Arachidonic acid can then be converted to hydroxyeicosatrienoic acid (HETE) acid derivatives, which are subsequently converted to the potent inflammatory mediator leukotrienes. The assay provides an accurate and convenient method for screening lipoxygenase inhibitors by assaying the hydroperoxides produced by incubation of lipoxygenase (5-LO, 12-LO or 15-LO) with arachidonic acid. The ability of any of the active ingredients, combinations of each of the ingredients, or compositions having the combinations disclosed in this specification to inhibit enzyme activity can be determined using a colorimetric LO inhibitor screening kit (#760700, Cayman Chemical). The purified 15-lipoxygenase and the test component can be mixed in the test buffer and incubated with shaking at room temperature for 10 minutes. After incubation, arachidonic acid may be added to start the reaction and the mixture may be incubated at room temperature for an additional 10 minutes. The catalysis can be stopped by the addition of a colorimetric substrate and the color change assessed by reading the fluorescence plate at 490 nm. The percent inhibition of lipoxygenase activity can be calculated by comparison to an untreated control to determine the ability of each active ingredient, combination of any one of the ingredients, or composition having the combination disclosed in the specification to inhibit purified enzyme activity.

And (3) elastase determination: from Molecular Probes (Eugold, Oregon, USA)The elastase assay (kit # E-12056) can be used as an in vitro enzyme inhibition assay to measure the inhibition of elastase activity by each of the active ingredients, combinations of any of the ingredients, or compositions with the combinations disclosed in the specification.The kit comprises soluble bovine cervical ligament elastin, which may be labeled with a dye, such that fluorescence of the conjugate is quenched. The non-fluorescent substrate can be digested by elastase or other proteases to produce highly fluorescent fragments. The resulting fluorescence enhancement can be monitored with a fluorescent microplate reader. The digestion product from the elastin substrate has an absorption maximum at about 505nm and a fluorescence emission maximum at about 515 nm. When in useWhen the elastase assay kit is used for screening elastase inhibitors, the peptide N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl ketone is used as a selective collective inhibitor of elastase.

Oil control determination: assays for measuring reduction of sebum secretion in sebaceous glands and/or reduction of sebum production in sebaceous glands can be performed by using standard techniques known to those of ordinary skill in the art. In one case, a forehead may be used. Each of the active ingredients, combinations of any of the ingredients, or compositions having the combinations disclosed in this specification can be administered once or twice daily to a portion of the forehead for a set number of days (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or more than 14 days), while other portions of the forehead are not treated with the composition. After the expiration of the set number of days, sebum secretion can be analyzed by applying a fine oil absorbing paper to the treated and untreated forehead skin. The analysis was performed by first removing all sebum from treated and untreated areas with wet and dry cloths. An oil absorbing paper may then be applied to the treated and untreated forehead area, and a rubber band may be placed around the forehead to gently press the oil absorbing paper against the skin. After 2 hours, the oil absorbing paper can be removed, allowed to dry and then transilluminated. A darker colored oil absorbing paper corresponds to more sebum secretion, while a lighter colored oil absorbing paper corresponds to reduced sebum secretion.

Determination of erythema: the assay to measure the reduction in skin redness can be evaluated using a Minolta colorimeter. Skin erythema can be induced by applying a 0.2% sodium lauryl sulfate solution to the forearm of a subject. The area was protected with a closed patch for 24 hours. After 24 hours, the patch is removed and a Minolta colorimeter a may be used^*The values were evaluated for redness induced by stimulation. a is^*Values measure the change in skin color in the red region. Immediately after the reading, the area is treated with any one of the active ingredients, active ingredient combinations, or compositions containing the combinations disclosed in the present specification. Repeated measurements may be made periodically to determine the ability of the formulation to reduce redness and irritation.

Determination of skin clarity and reduction of freckles and age spots: using a Minolta colorimeterSkin clarity and reduction of freckles and age spots were evaluated. The a-value of the Minolta chromameter can be used to assess skin color changes to determine the likelihood of irritation due to product handling. a is^*Values measure the change in skin color in the red region. This is used to determine whether each of the active ingredients, any combination of ingredients, or compositions containing the combination disclosed in this specification induces a stimulus. The measurements may be taken on each side of the face and averaged as the values for the left and right faces. Skin clarity can also be measured using a Minolta colorimeter. The measurements are a combination of a, b and L values from the Minolta chromameter and are related to the brightness of the skin and are closely related to the smoothness and hydration of the skin. Skin measurements were performed as above. In one non-limiting aspect, skin clarity can be described as L/C, where C is chroma and is defined as (a)²+b²)^1/2。

Skin dryness, surface fine lines, skin smoothness and skin tone measurements: dry skin, surface fine lines, skin smoothness and skin tone can be assessed using clinical scoring techniques. For example, the clinical score for dry skin can be determined by the five-point standard Kligman Scale: (0) the skin is soft and moist; (1) the skin appeared normal with no visible dryness; (2) the skin felt slightly dry to the touch without visible flaking; (3) the skin feels dry, rough and has a whitish appearance with some scaling; and (4) the skin feels very dry, rough and has a scaly whitish appearance. The assessments may be performed independently by two clinicians and averaged.

Skin color clinical score determination: clinical scoring of skin tone can be performed by a ten-point analog numerical scale: (10) smooth and even skin, pink brown color. There were no dark, red or peeled plaques when examined with a hand-held magnifying glass. The fine texture of the skin is very uniform to the touch; (7) the uniform skin color was observed without a magnifying glass. There was no peeling area, but there was slight discoloration due to pigmentation or erythema. No discoloration of greater than 1cm in diameter; (4) skin discoloration and uneven texture was easily noticed. And (5) peeling a little. Certain areas of the skin are rough to the touch; and (1) uneven skin coloration and texture. Peeling and discoloration in various areas, hypopigmentation, redness or black spots. Large areas of uneven coloration with diameters exceeding 1 cm. The assessments may be performed independently by two clinicians and averaged.

Clinical scoring determination of skin smoothness: the clinical score of skin smoothness can be analyzed on a ten point analog numerical scale: (10) smooth, the skin is moist and shiny, there is no resistance to the fingers brushing across the surface; (7) a certain degree of smoothness and slight resistance; (4) coarse, visibly changing, friction is present; and (1) rough, flaky, uneven surfaces. The assessments may be performed independently by two clinicians and averaged.

Determination of skin smoothness and wrinkle reduction by the method disclosed by packaman et al (1978): reduction of skin smoothness and wrinkles can also be visually assessed using the method disclosed by packaman et al (1978). For example, the depth, shallowness and total number of surface lines (SFLs) for each subject can be carefully scored and recorded at the time of visit. The numerical score is obtained by multiplying the number factor by the depth/width/length factor. The scores of the eye area and mouth area (left and right) are obtained and added together as the total wrinkle score.

Skin firmness measurements with a Hargens Ballistometer: skin firmness can be measured with a Hargens Ballistometer, which is a device that assesses skin elasticity and firmness by landing small objects on the skin and recording the first two rebound peaks. The Ballistometry used a small lightweight probe with a relatively blunt tip (4 square millimeters-contact area). The probe gently penetrates into the skin and measurements are obtained that depend on the properties of the outer layers of the skin, including the stratum corneum and the outer epidermis and parts of the dermis.

Skin softness/flexibility measurements with Gas Bearing electrodynameter: skin softness/flexibility can be evaluated using a Gas Bearing electrodynameter, an instrument that measures skin stress/strain properties. The viscoelastic properties of skin are related to skin moisturization. Measurement of the intended site of the cheek area may be achieved by adhering the probe to the skin surface with double sided tape. A force of about 3.5 grams force (gf) can be applied in parallel to the skin surface and accurately measure the displacement of the skin. The flexibility of the skin can then be calculated and expressed in DSR (dynamic spring rate, in units of gm/mm).

Visualization of lines and wrinkles with replica: the appearance of lines and wrinkles on the skin can be assessed using a replica, which is an impression of the skin surface. Materials such as silicone rubber may be used. The replica can be analyzed by image analysis. The change in line and wrinkle visibility can be objectively quantified by removing a silicon replica from the face of the subject and analyzing the replica image with a computer image analysis system. Replicas can be taken from the eye area and neck area and photographed with a digital camera at low illumination angles of incidence. The digital image may be analyzed with an image processing program and wrinkles or fine lines may be determined.

Skin surface profiling with profilometer/stylus method: the skin surface profile can be measured by using a profilometer/stylus method. This involves flashing or dragging a stylus across the replica surface. The vertical displacement of the stylus can be recorded into a computer by means of a distance sensor and, after scanning the replica over a certain distance, can be generated as a two-dimensional curved surface by analysis of the cross-section of the skin contour. The scan may be repeated any number of times along a fixed axis to produce a simulated 3D image of the skin. Ten random replica cross sections can be obtained using stylus technology and combined to produce an average. Values of interest include Ra, which is the arithmetic mean of all roughness (height) values calculated by integrating the profile height with respect to the mean profile height; rt, the maximum vertical distance between the highest peak and the lowest valley; and Rz, which is the average peak amplitude minus the average peak height. Values are given as values in mm. The device should be normalized by scanning a metal standard of known value before each use. The Ra value can be calculated by the following formula: r_aNormalized roughness; l_mTransverse (scan) length; and y is profile position relative to mean profile heightAbsolute value of degree (x-axis).

MELANODERM^TMAnd (3) determination: in other non-limiting aspects, the skin can be treated by using a skin mimic such as melandoderm^TMTo evaluate the efficacy of each of the active ingredients, combinations of any one of the ingredients, or compositions having the combinations disclosed in the specification. Melanocytes, which are one of the cells in skin mimics, are visibly colored when exposed to the precursor of melanin, L-dihydroxyphenylalanine (L-DOPA). Skin analog melandoderm^TMThe various substrates comprising each of the active ingredients disclosed in the specification, any combination of ingredients, or compositions having the combination, or the substrate alone may be used for treatment as a control. Alternatively, an untreated sample of a skin analog can be used as a control.

Keratinocyte monolayer permeability: the permeability change of the keratinocyte monolayer due to each active ingredient, any combination of ingredients, or a composition having the combination disclosed in the present specification can be determined. The permeability of the keratinocyte monolayer is a measure of the integrity of the skin barrier. By way of non-limiting example, keratinocyte monolayer permeability in treated and untreated keratinocytes can be determined using the in vitro vascular permeability assay of Millipore (ECM 642). This assay analyzes endothelial cell adsorption, transport and permeability. Briefly, adult epidermal keratinocytes from Life Technologies (C-005-5C) can be seeded onto porous collagen-coated membranes within the collection wells. At 37 ℃ and 5% CO₂In keratinocytes derived from Life TechnologiesCulturing in growth medium (M-EP-500-CA) for 24 hours, whereinThe growth medium contained calcium and was supplemented with keratinocyte growth factor (HKGS) from Life Technologies (S-101-5). This incubation time allowed the cells to form a monolayer and close the membrane pores. However, the device is not suitable for use in a kitchenThe medium was then replaced with fresh medium with (test sample) or without (untreated control) test compound/extract and incubated at 37 ℃ and 5% CO₂In which keratinocytes are incubated for a further 48 hours. After incubation with/without test compound/extract, to determine the permeability of the keratinocyte monolayer, the medium was replaced with fresh medium containing high molecular weight Fluorescein Isothiocyanate (FITC) -dextran and incubated at 37 ℃ and 5% CO₂The keratinocytes were incubated for a further 4 hours. During the 4 hour incubation, FITC can pass through the keratinocyte monolayer and porous membrane into the collection well at a rate proportional to the permeability of the monolayer membrane. After 4 hours incubation, cell viability and FITC content in the collection wells can be determined. For FITC content, the media in the collection well was collected and the fluorescence of the media was measured at 480nm (Em) under excitation at 520 nm. The percent permeability and percent change from untreated control can be determined by the following equation: percent permeability ═ ((Ex/Em average for test samples)/Ex/Em average for untreated controls) × 100; percent change-percent permeability of the test sample-percent permeability of the untreated control.

Production of hyaluronic acid: the change in hyaluronic acid production in human dermal fibroblasts due to each of any one of the active ingredients, the combination of any one of the ingredients, or the composition having the combination disclosed in the present specification can be measured. HA is a polysaccharide involved in the stability of the matrix structure, and it is also involved in providing turgor pressure to tissues and cells. As a non-limiting example, compounds from R may be used&Hyaluronan DuoSet ELISA kit (DY3614) by D Systems measures HA production in treated and untreated adult dermal fibroblast (HDFa) cells. In this analysis, for the generation of samples, prior to treatment, at 37 ℃ and 10% CO₂Next, sub-confluent HDFa cells (C-13-5C) obtained from Cascade Biologics were incubated in starvation medium (0.15% bovine fetal serum and 1% penicillin streptomycin solution in Dulbecco's modified Eagle medium) for 72 hours. Then using a positive control (from a test compound) containing the test compoundPhorbol 12-tetradecanoic acid 13-acetate from Sigma-Aldrich (P1585) and platelets derived from growth factors from Sigma-Aldrich (P3201)) or fresh starvation medium without added controls were cultured for 24 hours. The medium was then collected and frozen at-80 ℃ until used in the ELISA assay.

Briefly, the ELISA assay employs a quantitative sandwich ELISA technique, so that capture antibodies specific for HA can be pre-coated on a microplate. Standards, media from treated and untreated cells were pipetted into the microplate so that all HA present was bound by the immobilized antibody. After washing away all unbound material, an enzyme-linked detection antibody specific for HA is added to the wells. After washing to remove all unbound antibody-enzyme reagent, a substrate solution is added to the wells, allowing the color to develop in proportion to the amount of HA bound in the initial step. Color development is terminated at a specified time and the intensity of the color can be measured at 450nm using a microplate reader.

Inhibition of hyaluronidase activity: the change in hyaluronidase activity due to each of the active ingredients, the combination of any one of the ingredients, or the composition having the combination disclosed in the present specification can be determined. Hyaluronidase is an enzyme that breaks down HA. HA is a polysaccharide involved in the stability of the matrix structure, which is involved in providing turgor pressure to tissues and cells. As a non-limiting example, hyaluronic acid activity may be determined using an in vitro protocol modified by Sigma-Aldrich protocol # EC 3.2.1.35. Briefly, hyaluronidase type 1-S from Sigma-Aldrich (H3506) was added to reaction wells of microplates containing test compounds or controls. Tannic acid can be used as a positive control inhibitor, the enzyme of the control is not added with the test compound, and wells containing the test compound or the positive control but not hyaluronidase can be used as background negative controls. Wells were incubated at 37 ℃ for 10 min before addition of substrate (HA). Substrate was added and the reaction was incubated at 37 ℃ for 45 minutes. Then, a part of each reaction solution was transferred to a solution of sodium acetate and acetic acid at pH 3.75 and gently mixed to terminate the part of the reaction (stop well). After a portion of the reaction solution is added to the stop well, both the stop well and the reaction well should contain the same volume of solution. Both reaction and stop wells were incubated at room temperature for 10 minutes. The absorbance at 600nm of the reaction well and the stop well was then measured. The inhibition rate can be calculated using the following formula: inhibitor (or control) activity ═ (absorbance at 600nm inhibitor stop wells-absorbance at 600nm inhibitor reaction wells); initial activity-control enzyme absorbance at 600 nm; the inhibition rate is [ (initial activity/inhibitor activity) × 100] -100.

Activity of peroxisome proliferator-activated receptor gamma (PPAR- γ): changes in PPAR- γ activity due to each of the active ingredients, the combination of any of the ingredients, or the composition having the combination disclosed in the present specification can be determined. PPAR-gamma is an important receptor for sebum production. By way of non-limiting example, PPAR-gamma activity is determined using a biological assay that analyzes the ability of a test compound or composition to inhibit ligand binding. Briefly, a fluorescent small molecule Pan-PPAR ligand FLUORMONE from Life Technologies was used^TMPan-PPAR Green (PV4894) to determine whether a test compound or composition is capable of inhibiting ligand binding to PPAR- γ. The sample wells contain PPAR- γ and a fluorescent ligand, and: test compounds or compositions (test); reference inhibitor rosiglitazone (positive control); or no test compound (negative control). The wells are incubated for a set period of time to allow an opportunity for ligand binding to PPAR-gamma. The fluorescence polarization of each sample well can then be measured and compared to a negative control well to determine the inhibition of the test compound or composition.

Endothelial cell tube formation: the formation of endothelial cells is associated with angiogenesis and capillary formation. Capillary formation and vascularization can lead to redness of the skin and rosacea. The ability of endothelial cells to form tubes, in the presence or absence of test extracts and compounds, can be determined in a cell culture system using a capillary disruption assay with preformed primary Human Umbilical Vein Endothelial Cells (HUVECs).

Briefly, HUVECs are cultured in vitro in an extracellular matrix, which stimulates endothelial cell attachment and tubular morphogenesis to form a capillary-like luminal structure. These in vitro formed capillaries are similar in many respects to human vascular capillaries. Capillary tests are based on this phenomenon and are used to evaluate potential vasculature targeting agents.

At 5% CO₂HUVEC cultures were grown in cell culture vessels at 37 ℃. The complete growth medium for HUVEC was endothelial tube cell basal medium (EBM) supplemented with 2% Fetal Bovine Serum (FBS), 12. mu.g/ml bovine brain extract, 1. mu.g/ml cortisol and 1. mu.g/ml GA-1000 (gentamicin-amphotericin). HUVEC cultures between passage 3 and passage 8 can be used for all assays.

HUVECs were pre-labeled with the fluorescent reagent Calcein AM and seeded into extracellular matrix-coated 96-well culture plates containing complete growth medium. Approximately four hours after the morphogenetic process, endothelial capillaries formed. Then, as a treatment condition, a set dose of the test agent was administered to the formed capillary culture in a volume of 50 μ l. Vehicle or test agent may be added to the untreated control group. Sotan (Sutent) is an FDA-approved anti-angiogenic drug, one concentration of which may be included as a control for determining performance. About six hours after treatment, capillary destruction activity under the treatment conditions can be quantified by microscopic examination of the endothelial tubule morphology in each well, imaging it. Each test condition can be performed in duplicate wells, including controls.

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All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

Reference to the literature

The following references provide exemplary procedures or other additional details to the details set forth herein to the extent they are expressly incorporated by reference.

Cosmetic ingredient dictionary, third edition, CTFA, 1982.

International cosmetic ingredient dictionary, fourth edition, CTFA, 1991

International cosmetic ingredient dictionary and handbook, tenth edition, CTFA, 2004

International cosmetic ingredient dictionary and handbook, twelfth edition, CTFA, 2008

U.S. Pat. No. 5972993

U.S. publication No. 2015/0011489

Tighe et al (2013), Terphin-4-ol is the most active ingredient of tea tree oil to kill Demodex mite, Translational Vision Science & Technology,2(7), 1-8. DOI 10.1167/tvst.2.7.2