阅读说明：本技术 Uvb诱导的光损伤：用于局部治疗的组合物和方法 (UVB-induced photodamage: compositions and methods for topical treatment ) 是由 Z·本特维奇 J·梅洛尔 D·兰 G·科恩 于 2019-08-01 设计创作，主要内容包括：本发明涉及包含没药属植物的水基提取物的组合物,及其用于预防或治疗受试者皮肤的紫外线(ultra-violate)(UV)辐射损伤的使用方法。(The present invention relates to compositions comprising water-based extracts of commiphora genus plants and methods of use thereof for preventing or treating Ultraviolet (UV) radiation damage to the skin of a subject.)

1. A composition comprising an aqueous-based extract of a bisabolo plant.

2. A composition comprising an extract obtained by aqueous-based extraction of a plant of the genus commiphora.

3. The composition of claim 1 or 2, wherein the bisabolo plant comprises:

a. leaf: 10-30% (w/w) by dry weight;

b. fruit: 5-15% (w/w) by dry weight;

c. branch: 20-60% (w/w) by dry weight;

or any combination thereof.

4. The composition of any one of claims 1 to 3, wherein the composition comprises at least 10% (w/w) juice by dry weight.

5. The composition of any one of claims 1 to 4, wherein the composition comprises 0.2 to 2% (w/w) phenolic compounds.

6. The composition of any one of claims 1 to 5, wherein the composition comprises 0.01 to 0.05% (w/w) Volatile Organic Compounds (VOCs).

7. The composition of any one of claims 1 to 6, wherein the genus Commiphora is Coumacarat (Commiphora gileadensis).

8. The composition of any one of claims 1 to 7, wherein the composition comprises a portion of the aqueous-based extract.

9. The composition of any one of claims 1 to 8, further comprising an antioxidant, a chelating agent, a cleansing agent, a skin protectant, a sunscreen agent, a skin lightening agent, an anti-wrinkle agent, an anti-inflammatory agent, an anti-aging agent, or any combination thereof.

10. A composition comprising the composition of any one of claims 1 to 9 and a cosmetically acceptable carrier.

11. A method for preventing or treating Ultraviolet (UV) radiation damage to the skin of a subject comprising topically applying to the skin of the subject a composition comprising an effective amount of an aqueous-based extract of a commiphora plant.

12. The method of claim 11, wherein the bisabolo plant comprises:

a. leaf: 10-30% (w/w) by dry weight;

b. fruit: 5-15% (w/w) by dry weight;

c. branch: 20-60% (w/w) by dry weight;

or any combination thereof.

13. The method of claim 11 or 12, wherein the composition comprises at least 10% (w/w) juice by dry weight.

14. The method of any one of claims 11 to 13, wherein the composition comprises 0.2 to 2% (w/w) phenolic compounds.

15. The method of any one of claims 11 to 14, wherein the composition comprises 0.01 to 0.05% (w/w) VOC.

16. The method of any one of claims 11 to 15, wherein the commiphora is mecargonium.

17. The method of any one of claims 11 to 16, wherein the composition comprises a portion of the aqueous-based extract.

18. The method of any one of claims 11 to 17, wherein the composition is topically applied to the skin of the subject before, during, or after exposure to UV radiation.

19. The method of any one of claims 11 to 18, wherein the UV radiation is UVB radiation.

20. The method of any one of claims 11 to 19, wherein the subject is at risk of sun exposure.

21. The method of any one of claims 11 to 20, wherein the composition further comprises a cosmetically acceptable carrier.

22. The method of any one of claims 11 to 21, wherein the composition is in the form of a cream, lotion, ointment or spray.

23. The method of any one of claims 11 to 22, wherein the composition further comprises an antioxidant, a chelating agent, a cleansing agent, a skin protectant, a sunscreen, a skin lightening agent, an anti-wrinkle agent, an anti-inflammatory agent, an anti-aging agent, or any combination thereof.

Technical Field

The present invention, in some embodiments thereof, relates to plant extracts and uses thereof, such as for preventing or treating damage caused by Ultraviolet (UV) radiation.

Background

The incidence of melanoma and non-melanoma skin cancers reported worldwide has continued to increase over the last few decades. These are mainly due to the increased exposure to solar radiation and its intense Ultraviolet (UV) radiation. The latter induce direct damage to the DNA and membranes of cells and are therefore considered to be a major risk factor and cause of skin cancer. UVB-induced carcinogenesis is associated with UV absorption through cellular DNA, which results in damage to the DNA, producing mutagenic dimeric photoproducts (i.e., cyclobutane-pyrimidine dimer (CPD) and 6-4 photoproducts (6-4 PP)).

Sunscreens which attenuate UV radiation by absorption are the primary protectants against solar exposure. They protect the skin from radiation-induced photobiological changes and typically contain a combination of various active agents that absorb or scatter incident radiation, the various active agents being dispersed in various formulations. The active ingredient is not absorbed by the epidermis, dermis or into the blood. However, serious concerns have been raised about their ecotoxicity, photo-instability, phototoxicity and photo-allergic reactions, and these are still presentIs not solved. In a study involving 2,715 patients, sunscreens (especially benzophenone-3) were the most common cause of photosensitivity. Since its initial publication, this finding has been reconfirmed by a second study of over a thousand patients. Controlled human studies have shown that the use of sunscreens does not always prevent sunburn. One reason for this day is that the individual applied sunscreen at concentrations that are not comparable to those used in the controlled Sun Protection Factor (SPF) test (i.e., 2 mg/cm)²) The same is true. Other studies have shown that most sunscreen users only apply 25% to 75% of this amount. In addition, no treatment is currently available for reversing the sun damage caused by overexposure to UV radiation.

Currently, there is an increasing demand in western society for natural and less toxic sunscreens. Natural herbal products are receiving increasing attention and are a reliable alternative to synthetic sunscreens, especially those exhibiting antioxidant, UV absorbing, and/or anti-inflammatory properties. There remains a need for safe and effective compositions, particularly cosmetic and skin care compositions, that can be used to treat or prevent damage from solar radiation and maintain the viability of skin cells, while excluding potentially harmful synthetic compounds. For this reason, such protection is desired not only for healthy individuals, but also for patients with high sensitivity to light (photodermatitis), for which even short-term exposure to sunlight may lead to harmful effects ranging from erythema of the skin to cancer formation.

Disclosure of Invention

In some embodiments, the present invention relates to compositions comprising an aqueous-based extract of a Commiphora (Commiphora) plant. In some embodiments, the present invention relates to compositions comprising an extract obtained by aqueous-based extraction of a bisabolo plant. In some embodiments, the present invention relates to methods of preventing or treating Ultraviolet (UV) radiation damage to the skin of a subject using the compositions of the present invention. In some embodiments, the present invention is based, in part, on the following findings: aqueous extracts of maccaramelia (c. gileadensis) are highly effective as UV radiation protective agents.

According to one aspect, a composition is provided comprising an aqueous-based extract of a bisabolo plant.

According to another aspect, there is provided a composition comprising an extract obtained by aqueous-based extraction of a bisabolo plant.

In some embodiments, the composition comprises: (a) leaf: 10 to 30% (w/w) by dry weight; (b) fruit: 5 to 15% (w/w) by dry weight; (c) branch: 20 to 60% (w/w) by dry weight; or any combination thereof. In some embodiments, the composition comprises at least 10% (w/w) juice (sap) by dry weight.

In some embodiments, the composition comprises 0.2 to 2% (w/w) phenolic compound. In some embodiments, the composition comprises 0.01 to 0.05% (w/w) Volatile Organic Compounds (VOCs).

In some embodiments, the genus Commiphora is meconium lipid (Commiphora gileadensis).

In some embodiments, the composition comprises a portion (fraction) of the aqueous-based extract.

In some embodiments, the composition further comprises an antioxidant, a chelating agent, a cleansing agent, a skin protectant, a sunscreen agent, a skin lightening agent, an anti-wrinkle agent, an anti-inflammatory agent, an anti-aging agent, or any combination thereof.

In some embodiments, the composition comprises a dermatologically acceptable carrier. In some embodiments, the composition comprises a cosmetically acceptable carrier. In some embodiments, the composition is in the form of a cream, lotion, ointment, or spray.

In some embodiments, the composition is topically applied to the skin of the subject before, during, or after exposure to UV radiation. In some embodiments, the UV radiation is UVB radiation. In some embodiments, the subject is at risk of sun exposure.

According to another aspect, there is provided a method of preventing or treating Ultraviolet (UV) radiation damage to the skin of a subject comprising topically applying to the skin of the subject a composition comprising an effective amount of an aqueous-based extract of a bisabolo plant.

Unless defined otherwise, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, the following description of exemplary methods and/or materials is provided. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be necessarily limiting.

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

Drawings

FIGS. 1A-1B are gas chromatographic analyses of WSE (1A) and WPE (IB) of mecamylene.

Figure 2 is a vertical bar graph showing the phenol content (grey bars) and antioxidant capacity (black bars) of whole juice emulsions (WSE) and whole plant emulsions/extracts (WPE) of macchia.

Figures 3A-3E are vertical histograms and examples demonstrating that WPE and WSE of mecamylaxanthus attenuate UVB-induced apoptosis in human skin explant organ culture (HSOC). (3A) Is a vertical bar graph showing the photoprotective activity of mecaxane over the 7 month period. WSE (0.8 mg/cm) for HSOC²) And WPE (0.9 mg/cm)²) Local pretreatment took 24 hours. After incubation, with UVB (400 mJ/cm)²) The explant is challenged. Caspase-3 activity levels were measured 24 hours after irradiation. (3B) Is a vertical bar graph showing the cytotoxicity of mecaxane over the 7 month period. HSOC was locally pretreated with WSE and WPE for 24 hours. The skin is then exposed to UVB. Viability was measured 24 hours after irradiation (MTT assay). Values are mean ± SEM; n is 4. (3C) Is from 7 months to 8 monthsNon-limiting schematic of the changes in effect caused by the maccarael WPE. It appears that the activity of the macchian WPE is positively switched between the first and the second half of 7 months. (3D) Is a vertical bar graph showing that WPE is dose-dependent on the photoprotective effect of reduced apoptosis in HSOCs. (3E) Is a vertical bar graph showing the differential active components of WPE. The stem of the plant was removed, the hard bark was separated from the pith, and apoptosis of HSOC was assessed as described above. It was shown that the main reservoir of the substances responsible for photoprotective activity and cytotoxicity is located in the bark of the plants, not in the pith.

Fig. 4A-4B are vertical bar graphs showing that maccarat retains its photoprotective ability under multiple exposures. The WSE and WPE of maccarat retained their photoprotective activity after repeated exposure of HSOC to harmful sunlight. With WSE (0.8 mg/cm)²)、WPE(0.9mg/cm²) And WSE/WPE 1: the HSOC was locally pretreated with the mixture of 1(v/v) for 24 hours. The HSOC was then exposed to UVB. The next day, HSOC was irradiated again. Apoptosis (4A) and viability (4B) were measured 24 hours after irradiation. Values are mean ± SEM; n is 4. Indicates relative to control P without pretreatment<0.05; # indicates control P relative to UVB irradiation<0.05。

FIGS. 5A-5B are vertical bar graphs showing that the macchia balsam obtained at Ein-Gedi (07/08/2016) in Israel proved to be advantageous in comparative analysis over other available ethereal myrrh oils. The WPE and WSE of native mugatran were shown to be more effective than etheric myrrh oils from other regions of the earth. HSOC was locally pretreated for 24 hours using WSE, WPE and other commercial products. Left side of the graph: control, WSE and WPE; on the right side of the graph: commercial, ethereal oil of the aegybia and vienna. The HSOC was then exposed to UVB. Apoptosis (5A) and viability (5B) were measured 24 hours after irradiation. Values are mean ± SEM; n is 4. Indicates P <0.05 relative to control without pretreatment; # indicates that P <0.05 relative to control UVB irradiation.

FIGS. 6A-6D are vertical histograms showing that the lipid obtained at Ein-Gedi (07/08/2016) in Israel proved to be advantageous compared to commercial sunscreens. The HSOC was topically pretreated with cream containing UV-blocker ZnO (zinc oxide), WSE or WPE for 24 hours. Left side of the graph: control and cream-zinc oxide; in the middle: cream-WSE; right side: cream-WPE. The HSOC was then exposed to UVB. Apoptosis (6A) and viability (6B) were measured 24 hours after irradiation. Topical WPE and WSE of mecargane are less cytotoxic to HSOC than the most popular commercial sunscreens. HSOC was pre-treated with WPE and mineral sunscreens with various sun protection factors (SPF; 15 to 100) for 24 hours. Left side of the graph: control and cream-WPE; right side: a sunscreen product. The HSOC was then exposed to UVB. Apoptosis (6C) and viability (6D) were measured 24 hours after UVB. Values are mean ± SEM; n is 4. Indicates P <0.05 relative to control without pretreatment; # indicates that P <0.05 relative to control UVB irradiation.

Figures 7A-7C are vertical bar graphs showing that the addition of WPE and WSE of native maccarat to a sunscreen emulsion results in a strong photoprotective synergy. HSOCs pretreated with mugwort WPE and WSE and sunscreen emulsions showed lower levels of UVB-induced apoptosis. Various concentrations of the maccaramels emulsion were mixed with a sunscreen emulsion (30 SPF). HSOC was topically pretreated with the sunscreen emulsion-emulsion made for 24 hours. Left side of the graph: control, WPE and original sunscreen emulsion spray (LS); in the middle: 25% mixture of LS and WPE; right side: 10% mixture of LS and WPE. The HSOC was then exposed to UVB. Apoptosis (7A) and viability (7B) were measured 24 hours after irradiation. Values are mean ± SEM; n is 4. Indicates P <0.05 relative to control without pretreatment; # indicates that P <0.05 relative to control UVB irradiation. (7C) HSOC pretreated with mujiaxiang WPE and WSE and sunscreen emulsions showed lower levels of UVB induced photodamage. Analysis of CPD formation in freshly prepared emulsions subjected to mecahumarol (16/08/2016) and subsequent exposure to UVB radiation in epidermal cells. Dimer formation was measured by ELISA. HSOC was topically pretreated with the manufactured emulsion-cream for 24 hours. Left side of the graph: control, emulsion (WSE and WPE); right side: original sunscreen emulsion spray (LS) and a mixture of LS and 10% emulsion. The HSOC was then exposed to UVB. The irradiation was followed by testing for DNA photodamage. Values are mean ± SEM; n is 4. Indicates P <0.05 relative to control without pretreatment; # indicates that P <0.05 relative to control UVB irradiation.

Figures 8A-8D show that the mogroside WPE and WSE have dual anti-inflammatory effects. (8A-8C) are vertical histograms showing that the Lipocalin WPE and WSE down-regulate pro-inflammatory cytokine production in HSOC. After topical application of the mecamylaxane WPE and WSE to the HSOC inflammatory model (i.e., HSOC that had been previously exposed to LPS (lipopolysaccharide) and EGF (epidermal growth factor)) and incubated for 48 hours, the media was harvested for ELISA assays to determine the amounts of the proinflammatory cytokines IL-6(8A), IL-8(8B) and TNF α (8C) in HSOC media. Values are mean ± SEM; n is 4. Indicates P <0.05 relative to untreated control; # indicates that P <0.05 relative to control induced by LPS plus EGF. (8D) Is a vertical bar graph of Western blot analysis of heme oxygenase-I (HO-1) and corresponding densitometry, showing that Michael balsam WPE and WSE increase HO-1 enzyme expression. Western blot analysis of HO-1 expression in HSOC after pro-inflammatory stimulation with EGF plus LPS was performed using primary anti-HO-1 antibody; anti- β -actin was used as a loading control. Histogram shows densitometric analysis of HO-1 signal normalized with β -actin. Image analysis was done by ImageJ software. The level of HO-1 in HSOC was assessed after induction of inflammation by EGF and LPS and 48 hours of culture with addition of WPE or WSE followed by analysis. Values are mean ± SEM; n is 3. Indicates P <0.05 relative to control without pretreatment; # shows that the control P induced relative to EGF + LPS is < 0.05.

Detailed Description

In some embodiments, the present invention relates to compositions comprising an aqueous-based extract of a bisabolo plant. In some embodiments, the present invention relates to compositions comprising an extract obtained by aqueous-based extraction of a bisabolo plant. In some embodiments, the present invention relates to methods of preventing or treating Ultraviolet (UV) radiation damage to the skin of a subject using the compositions of the present invention.

Extracts and compositions

In some embodiments, the present invention relates to plant extracts. In some embodiments, the plant extract of the present invention is derived from a member of the genus Commiphora (also known as "myrrh"). In some embodiments, the member of the genus commiphora is meconium.

As used herein, an extract of the present invention includes the entire extract, a portion thereof, a compound isolated therefrom, or any combination thereof.

In some embodiments, the plant extract of magadipine is a solvent-based extract obtained from any selected portion of magadipine by a solvent extraction method. In some embodiments, the solvent according to the present invention is a polar solvent. In some embodiments, the polar solvent is water or any aqueous solution, such as a water-based buffer or culture medium. Non-limiting examples of aqueous-based buffers include, but are not limited to, Phosphate Buffered Saline (PBS), Tris, 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (HEPES), 3- (N-morpholino) propanesulfonic acid (MOPS), all of which will be apparent to one of ordinary skill in the art. In some embodiments, the polar solvent is compatible with mammalian skin.

In some embodiments, the plant material used in the extraction process comprises whole plants. In some embodiments, the plant material used in the extraction process comprises one or more different tissues from the plant. In some embodiments, the plant material used in the extraction process comprises leaves, stems, fruits, roots, juices, or any combination thereof.

In one embodiment, the composition comprises a plant material comprising leaves. In one embodiment, the number of leaves in the plant material on a dry weight basis is 5-20% (w/w), 10-35% (w/w), 15-40% (w/w), 20-45% (w/w), 30-55% (w/w), 40-60% (w/w), 50-70% (w/w), or 45-75% (w/w) of the composition on a dry weight basis. Each possibility represents a separate embodiment of the invention. In one embodiment, the composition comprises plant material comprising fruit. In one embodiment, the number of fruits in the plant material is 1-10% (w/w), 5-15% (w/w), 8-20% (w/w), 12-25% (w/w), 10-35% (w/w), 17-30% (w/w), or 20-40% (w/w), on a dry weight basis. Each possibility represents a separate embodiment of the invention. In one embodiment, the composition comprises plant material comprising tree branches. In one embodiment, the number of branches in the plant material is 1-5% (w/w), 4-10% (w/w), 8-18% (w/w), 10-30% (w/w), 15-25% (w/w), 20-28% (w/w), or 25-35% (w/w), on a dry weight basis. Each possibility represents a separate embodiment of the invention. In one embodiment, the composition comprises a plant material comprising juice. In one embodiment, the amount of juice in the plant material is 1-10% (w/w), 5-20% (w/w), 8-25% (w/w), 20-40% (w/w), 30-55% (w/w), 50-70% (w/w), 65-90% (w/w), or 85-99% (w/w), on a dry weight basis. Each possibility represents a separate embodiment of the invention.

As defined herein, the term "juice" refers to a fluid transported in the xylem vessels or phloem cells of a plant. In some embodiments, the juice is collected from fresh shoots. In some embodiments, the juice collected from the fresh shoots is centrifuged and the supernatant is collected and dissolved in a water-based solvent. In some embodiments, the juice is collected from the dried resin. In some embodiments, the dried resin is collected and washed. In some embodiments, the dried resin is ground into a powder. In some embodiments, the juice is extracted from a resin grind dissolved in a water-based solvent.

In one embodiment, "water-based" is a composition comprising at least 80% w/w water. In one embodiment, "water-based" is a composition comprising at least 85% w/w water. In one embodiment, "water-based" is a composition comprising at least 90% w/w water. In one embodiment, "water-based" is a composition comprising at least 95% w/w water. In one embodiment, "water-based" is a composition comprising at least 97% w/w water. In one embodiment, "aqueous-based" is an aqueous composition.

In some embodiments, the plant material is extracted immediately after harvesting. In some embodiments, the plant material is first dried and then extracted. In some embodiments, the plant material is first stored and then extracted. In some embodiments, the plant material is first treated and then stored. As used herein, processing prior to storage includes, for example, freezing, drying, lyophilizing, or any combination thereof. In some embodiments, the storage period is days to weeks, weeks to months, months to years, or any range therebetween. In some embodiments, the plant material is kept protected from light prior to storage, after storage, or both.

In some embodiments, the plant material is further processed prior to the extraction procedure to facilitate the extraction procedure. In some embodiments, processing methods prior to extraction include, but are not limited to, crushing, slicing, or shredding (e.g., by using a grinder or other device to break the plant parts into pieces or powder). In some embodiments, the processing method prior to extraction includes, but is not limited to, washing (e.g., by water, buffer, preservative-containing buffer, or acidic solution).

In some embodiments, the volume or amount of solvent used in the extraction procedure is proportional to the volume or amount of solid plant material. In one embodiment, the ratio is weight/weight (w/w). In some embodiments, the amount of solvent used in the extraction procedure ranges from the amount of solid plant material x1 to x100 (mass/mass). In some embodiments, the range of x1 to x100 (mass/mass) is x1 to x10 (mass/mass), × 5 to x 50 (mass/mass), × 2 to x40 (mass/mass), × 3 to x80 (mass/mass), × 7 to x25 (mass/mass), × 1 to x 20 (mass/mass), × 10 to x90 (mass/mass), × 15 to x85 (mass/mass), × 4 to x30 (mass/mass), x25 to x60 (mass/mass), x40 to x90 (mass/mass), x60 to x85 (mass/mass), x80 to x100 (mass/mass), or any range therebetween. Each possibility represents a separate embodiment of the invention.

In some embodiments, the extraction procedure comprises incubating the plant material for a period in the range of 5 minutes to 24 hours. In some embodiments, within the range of 5 minutes to 24 hours includes 5 minutes to 30 minutes, 20 minutes to 60 minutes, 1 hour to 3 hours, 2 hours to 5 hours, 4 hours to 10 hours, 8 hours to 16 hours, 12 hours to 18 hours, 15 hours to 20 hours, 19 hours to 24 hours, or any range therebetween. In some embodiments, the extraction procedure comprises incubating the plant material at a temperature in the range of 4 ℃ to 90 ℃. In some embodiments, in the range of 4 ℃ to 90 ℃ includes 4 ℃ to 30 ℃,5 ℃ to 40 ℃, 10 ℃ to 70 ℃, 30 ℃ to 65 ℃,50 ℃ to 85 ℃, 55 ℃ to 75 ℃, 60 ℃ to 80 ℃, 40 ℃ to 70 ℃, 10 ℃ to 50 ℃, or any range therebetween. Each possibility represents a separate embodiment of the invention.

In some embodiments, after the extraction procedure, the extracted liquid fraction is separated from the solid (insoluble) material. Separation of the liquid and solid portions is accomplished by one or more standard separation processes known to those of ordinary skill in the art and includes, but is not limited to, various centrifugation processes, filtration processes, or any combination thereof. In some embodiments, the extracted liquid portion is further subjected to one or more purification steps. Non-limiting examples of purification methods include, but are not limited to, solid-liquid extraction, liquid-liquid extraction, solid-phase extraction (SPE), membrane filtration, ultrafiltration, dialysis, electrophoresis, solvent concentration, centrifugation, ultracentrifugation, liquid or gas chromatography with or without high pressure (including size exclusion, affinity, etc.), lyophilization, evaporation, precipitation using various "carriers" (including polyvinylpolypyrrolidone (PVPP), carbon, antibodies, and the like), supercritical fluids (such as CO), and the like₂) Or a combination thereof.

In some embodiments, the extracts of the present invention are formulated into compositions for topical administration. In some embodiments, the extract is formulated in a form selected from the group consisting of: gels, foams, creams, ointments, emulsions, powders, suspensions or sprays. Each possibility represents a separate embodiment of the invention.

In some embodiments, the compositions of the present invention further comprise an acceptable cosmetic agent, as known in the art. In some embodiments, the cosmetic agent is selected from: xanthine, retinoid, alpha-hydroxy acid, beta-hydroxy acid, an a-2 adrenergic inhibitor, beta-adrenergic agonist, aromatase reaction inhibitor, antiestrogen, hydroquinone, ascorbic acid, kojic acid, corticosteroid, mucopolysaccharide, collagen, estrogen, isoflavone, cinnamic acid, benzoyl peroxide, tropolone, catechol, mercaptoamine, nicotinamide, tocopherol, ferulic acid, azelaic acid, botulinum, urea, a derivative, a salt thereof, or any combination thereof. Each possibility represents a separate embodiment of the invention.

In some embodiments, the composition further comprises a cosmetically acceptable diluent, carrier, or excipient. In one embodiment, the carrier comprises a liposome. In one embodiment, the carrier comprises micelles. In one embodiment, the carrier comprises microcapsules. In one embodiment, the carrier comprises any combination of liposomes, micelles, or microcapsules.

In some embodiments, the compositions further comprise acceptable additives conventionally used in cosmetic and dermatological formulations known to those skilled in the art.

In some embodiments, the composition further comprises an antioxidant, a chelating agent, a cleansing agent, a skin protectant, a sunscreen agent, a skin lightening agent, an anti-wrinkle agent, an anti-inflammatory agent, an anti-aging agent, or any combination thereof.

As used herein, "anti-inflammatory agent" refers to any component that helps to inhibit or suppress inflammation on or in or near the skin in bodily tissues, and thereby reduce redness and swelling of the skin. Non-limiting examples of anti-inflammatory components include, but are not limited to, vitamin E or its derivatives, zinc, allantoin, glycyrrhetinic acid (glycyrrhetic acid), azulene, mefenamic acid (mefenamic acid), phenylbutazone (phenylbutazone), indomethacin, ibuprofen, ketoprofen, epsilon-aminocaproic acid, hydrocortisone, panthenol or its derivatives or salts, zinc oxide, and diclofenac sodium.

In some embodiments, the composition further comprises one or more antioxidants. In some embodiments, the antioxidant comprises an enzymatic or non-enzymatic antioxidant. Non-limiting examples of enzymatic antioxidants include, but are not limited to, superoxide dismutase (SOD), catalase, and glutathione peroxidase. Non-limiting examples of non-enzymatic antioxidants include, but are not limited to, vitamin E (tocopherol) or its derivatives, vitamin a (retinol), vitamin C (ascorbic acid), carotenoids or its derivatives, beta-carotene, canthaxanthin, zeaxanthin, lycopene (lycopen), lutein, crocetin, capsanthin echinacoside (capsanthin echinacoside), caffeoyl derivatives, oligomeric or proanthrolols (proanthanols), green tea polyphenols, dibutyl hydroxytoluene, butyl hydroxyanisole, tannins or their derivatives, gallic acid, ellagic acid, flavonoids, flavones, catechins, quercetin, anthocyanins, quinones, ubiquinones, vitamin K, thiamine or its salts, riboflavin and riboflavin acetate, pyridoxine hydrochloride, pyridoxine dicaprylate (pyridoxine dicantanate), nicotinic acid, nicotinamide (niacin), Benzyl nicotinate, bihirubin, mannitol, tryptophan, histidine and nordihydroguaiaretic acid (nordihydroguaiaretic acid).

In some embodiments, the composition further comprises vitamin C in the form of ascorbyl palmitate, dipalmitate L-ascorbate (dipalmitate L-ascorbate), sodium L-ascorbate-2-sulfate (sodium L-ascorbate-2-sulfate), or an ascorbate salt (such as sodium, potassium, or calcium, or a mixture thereof). In some embodiments, the composition further comprises vitamin C in an amount ranging from 0.1 to 50% (w/w). In some embodiments, the composition further comprises vitamin a in the form of vitamin a palmitate. In some embodiments, the composition further comprises vitamin a in an amount ranging from 0.5 to 15% (w/w). In some embodiments, the composition further comprises one or more carotenoids, derivatives thereof or any mixture thereof in an amount ranging from 0.1 to 5% (w/w).

In some embodiments, the compositions of the present invention further comprise compounds having sunscreen and/or sunblock properties. Non-limiting examples include, but are not limited to, titanium dioxide, zinc oxide, talc, red petrolatum (red petrolatum), cinnamates (e.g., octyl methoxycinnamate), benzones (e.g., oxybenzone or 2-hydroxy-4-methoxybenzophenone), salicylates (e.g., salicylate or octyl salicylate), benzoic acids (e.g., p-aminobenzoic acid), and benzophenones (e.g., oxybenzone). The exact amount of sunscreen used in the composition will vary depending on the degree of protection desired from the UV radiation of sunlight and can be readily determined by one of ordinary skill in the art.

In some embodiments, the present invention relates to compositions comprising an effective amount of an aqueous-based extract of a commiphora plant for treating and/or preventing damage to skin cells or skin tissue of a subject caused by UV radiation.

Non-limiting examples of skin damage include, but are not limited to, the formation of wounds on the skin, unbalanced skin microbiome, redness, wrinkles, lesions, and others.

In some embodiments, the present invention relates to compositions for reducing inflammation of UVR-damaged skin. In some embodiments, the composition is for use in wound healing. In some embodiments, the compositions are used in dermatological or cosmetic procedures. In some embodiments, the composition is used in an anti-aging procedure. In some embodiments, the composition is for use in skin microbiome balancing.

The term "dermatological," as used herein with respect to the methods, procedures, and compositions used thereof, encompasses therapeutic agents or cosmetics, or both.

Methods of treatment or prevention

In some embodiments, the present invention relates to a method for preventing or treating Ultraviolet (UV) radiation damage to the skin of a subject comprising topically applying to the skin of the subject a composition comprising an effective amount of an aqueous-based extract of a bisabolo plant.

As used herein, "UV radiation damage" refers to any damage to any of the cells in the skin layer due to exposure to the ultraviolet radiation described herein. In some embodiments, the injury to the cell comprises: damage to DNA, RNA, proteins, membranes, metabolites, or any combination thereof. In some embodiments, the damage to the cell results in apoptosis, necrosis, or death of any type of cell. In some embodiments, the damage to the cell comprises a cancerous transformation of the cell. As defined herein, "cancerous transformation" refers to the onset of cancer in damaged cells. As used herein, "cancer" encompasses diseases associated with cell proliferation.

In some embodiments, the UV radiation damage causes a skin disease. In some embodiments, the skin disease comprises cancer. In some embodiments, the cancer comprises melanoma. In some embodiments, the disease comprises an photodermatosis. In some embodiments, the disease comprises Actinic Keratosis (AK).

As used herein, the term "treatment" of a disease, disorder or condition encompasses the reduction of at least one symptom thereof, the reduction of the severity thereof, or the inhibition of the progression thereof. Treatment need not mean that the disease, disorder or condition has been completely cured. To be an effective treatment, a composition useful herein need only reduce the severity of a disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide an improvement in the quality of life of a patient or subject.

As used herein, the term "prevention" of a disease, disorder or condition encompasses the delay, prevention, suppression or inhibition of the onset of the disease, disorder or condition. As used in accordance with the presently described subject matter, the term "prevention" relates to a prophylactic process in which a subject is exposed to a presently described composition prior to induction or onset of a disease/condition process. The term "suppression" is used to describe a condition in which the disease/disorder process has begun but no apparent symptoms of the condition have occurred. Thus, the cells of an individual may have a disease/disorder, but no external signs of the disease/disorder have been clinically recognized. In either case, the term prophylaxis is applicable to encompass both prophylaxis and suppression. Conversely, the term "treatment" refers to the clinical application of an active agent to combat an already existing condition, the clinical manifestations of which have been recognized in a patient.

As used herein, the term "subject" refers to an animal, more particularly to non-human mammals and human organisms. Non-limiting examples of non-human animals include: horses, cattle, camels, goats, sheep, dogs, cats, non-human primates, mice, rats, rabbits, hamsters, guinea pigs, pigs. In one embodiment, the subject is a human. In some embodiments, the subject in need thereof is a subject having and/or at risk of having a condition associated with a skin disease. In one embodiment, the skin disease is induced by exposure to UV radiation.

As used herein, the term "condition" includes anatomical and physiological deviations from normality that constitute an impairment to the normal state of a living animal or one of its parts, the impairment disrupting or altering the performance of bodily functions.

Unless otherwise indicated, any concentration range, percentage range, or ratio range recited herein is to be understood as including the concentration, percentage, or ratio of any integer within the range and fractions thereof (e.g., tenths and hundredths of integers).

Unless otherwise indicated, any numerical range recited herein in connection with any physical characteristic (e.g., weight) is to be understood as including any integer within the recited range.

In the discussion, unless otherwise specified, adjectives (e.g., "substantially" and "about" modify a condition or relational feature of one or more features of an embodiment of the invention) are to be understood to mean that the condition or characteristic is defined within a tolerance that is acceptable for operation of the embodiment for the intended application. Unless otherwise indicated, the word "or" in the specification and claims is considered to be an inclusive "or" rather than an exclusive "or" and indicates at least one or any combination of the items it incorporates.

It is to be understood that the terms "a" and "an," as used above and elsewhere herein, refer to "one or more" of the enumerated components. It will be clear to those of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Thus, the terms "a" and "an" and "at least one" are used interchangeably herein.

For the purpose of better understanding the present teachings, and in no way limiting the scope of the present teachings, all numbers expressing quantities, percentages, or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term "about," unless otherwise indicated. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

In the description and claims of this application, each of the verbs "comprise," "include," and "have," and their conjugates, are used to indicate that the object or objects of the verb are not necessarily a complete list of components, elements, or parts of the subject or subjects of the verb.

Other terms used herein are intended to be defined by their well-known meanings in the art.

Other objects, advantages and novel features of the present invention will become apparent to one of ordinary skill in the art upon examination of the following examples, which are not intended to be limiting. Additionally, as described above and in the claims section that follows, each of the various embodiments and aspects of the present invention are supported experimentally in the following examples.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiment is inoperable without those elements.

Examples

Materials and methods

Whole plant emulsion/extract (WPE) — "whole plant extract" is an emulsion that contains the entire bark and medulla (core) of the plant and is produced when the oil-gum-resin and all other parts of the plant are mixed with water and heated. WPE was obtained using either wet-treated fresh plants (wet process) or dry plants using dry process treatment.

Dry process-all parts of the plant (branches, leaves, fruits) are cut upside down in dark, warm and ventilated space, and the resulting dried plant is then ground and pulverized to obtain a powder. Next, the dried plant material was put together in a ratio of about 50% leaves, 30% fruits, and 20% branches, and washed with water and 1% (v/v) acetic acid. The material was left at room temperature for 15 minutes and then placed in a dark and ventilated space and cut into manageable pieces. The dried plant material is then ground to obtain a powder. Water was then added to the powder as follows: 3 volumes of wet plant material-1 volume of dry material, and 1.2kg of WPE could be obtained from 1kg of wet plant. For 50gr dry plant powder, 160ml water was added; and then incubated at 60 ℃ for one hour (with frequent stirring), and then cooled. After cooling, the liquid was filtered and then centrifuged at 1,500RPM for 15 minutes. The resulting precipitated material was then subjected to compression to expel the oil, gum and extract and then resuspended in 160ml of water.

Wet process-first, the plants are washed and cut into manageable pieces, and then an amount of water corresponding to the desired extract potency is added. Next, a pulverizer (juicer) was used to obtain a ground mixture having a fine structure. The semi-liquid mixture was then incubated at 60 ℃ for 1 hour, followed by cooling. After cooling, the mixture was subjected to compression to expel the oil, gum and extract and then filtered. Since the mixture is sensitive to light, it must be kept in a dark container.

WSE-whole juice emulsion (extract) -after wounding of the bisabolo plants, a clear concentrated liquid is exuded from the wound (pre-sap or CgSE) immediately, which then becomes cloudy and thicker (emulsion-like thick liquid). If it is left on the tree (or left to drip to the ground), it becomes a hard "cream-like" resin. The hardened resin (Sap) was then washed, cleaned and ground to a powder of 100 microns, and then filtered to obtain a powder. Water with or without preservatives is added to the powder, so that 3 to 5 litres of emulsion are obtained from a solution of 1kg of Sap. The weight of water was calculated as (Sap weight. times.3). times.1.2. The emulsion was incubated at 60 ℃ for 1 hour (with frequent stirring) and then centrifuged at 1,500RPM for 15 minutes. The supernatant was then filtered and the resulting WSE was sealed with beeswax and thawed, transferred to a container covered with aluminum foil and stored at-80 ℃ in the dark.

Cream formulations

As described herein, several creams containing active ingredients can be prepared and evaluated for their effectiveness in skin photoprotection before and after exposure to UVB. Zinc oxide (ZnO) cream-75% DDW + 14% ZnO + 7% sweet almond oil + 4% sibiral (Sepigel) -305. WPE cream-73.14% DDW + 17.31% WPE + 5.7% sweet almond oil + 3.85% sibirak-305. WSE cream-66.13% DDW + 16.67% WSE + 12.5% sweet almond oil + 4.7% sibiram-305.

Human ex vivo skin explant in organ culture (HSOC)

Intact human skin explants in organ culture (HSOC) have been used as experimental models of intact human skin. Human skin was obtained from adult healthy donors (female; 20-60 years old) undergoing abdominal reduction surgery. All experiments were performed under approval by the institutional review board (ethical "helsinki" board) of the Soroka medical center, Be' er-Sheva, israel. Skin explants were used no more than 12 hours after surgery. Briefly, the samples were cut into 0.8X 0.8cm sections using a mechanical microtome (designated pressing device)²The sheet of (1). Placing the explant in a sterile 6-well plate; in a high glucose DMEM medium (Life Technologies Ltd.,having 4,500mg/L D-glucose + L-glutamine, w/o sodium pyruvate, cat # s: 41965 039), the epidermis side is exposed to air. At 37 ℃ and 5% CO₂Explants were used after overnight recovery.

Ultraviolet B (UVB) radiation challenge for HSOC

Each experiment consisted of quadruplicates for each treatment. WPE, WSE or a combination of both are topically applied to the epidermis (0.9 or 0.8mg/cm, respectively)²The outer surface of the skin patch); in addition, to determine the efficacy of WPE, dose response analyses were performed with reduced WPE concentrations. WPE was diluted in water and applied at various concentrations on HSOC. After 24 hours incubation, the growth medium was aspirated and replaced with phosphate buffered saline (PBS × 1). A patch of skin (HSOC) was administered in a dose-responsive manner (400-750 mJ/cm)²) Exposure to UVB radiation challenge. Subsequently, PBS × 1 was aspirated and replaced with fresh medium. The HSOC was then allowed to recover for 24 hours under standard conditions. To collect the epidermis, the skin was incubated in PBS × l at 56 ℃ for 1 minute, after which the epidermis was physically separated from the dermis using forceps and a scalpel, and washed again in PBS × l. The epidermis was tested using the techniques described below.

Time course analysis was performed as follows: HSOC was cut and pretreated the day and placed in incubation; irradiating after 24 hours, and returning to incubate until collective harvest day; the (non-) pretreated controls were common to all irradiation groups.

Measurement of apoptosis by caspase-3 Activity assay

UVB-induced apoptosis in epidermal cells was determined using a caspase-3 enzyme activity fluorescence assay. The epidermal sheets were placed in 96-well plates and 125. mu.L of caspase-3 specific substrate solution (500mM DTT, 10% Triton X-100, caspase-3 substrate diluted 1: 1,000 in PBS. times.l) was added to each well. The fluorescence product of the enzyme was measured 20 times at 2 minute intervals using a microplate reader according to the supplier's instructions (37 ℃ C.; 40 minutes; Ex.355nm; Em.460nm, Calbiochem, Ltd., catalog No. 235425).

To exclude the possibility of WPE, WSE, pith extract or bark extract interacting directly and inhibiting the proteolytic activity of caspase-3, human recombinant C-terminal histidine-tagged caspase-3 enzyme: (Israel) with WPE, WSE, pith extract or bark extract and then assayed in 96-well multi-well plates (50 ng/well) using caspase-3 Substrate (Substrate) II, Fluorogenic to determine the fluorescent product of the enzyme. As a control, the known irreversible pan (pan) -caspase inhibitor (CFI) was used. The assay was performed at 37 ℃ for 40 minutes in a total volume of 100. mu.L.

Determination of viability

HSOC viability assay-by using the MTT assay (Thiazolyl Blue Tetrazolium Bromide),the directory number: m5655) determination of epidermal viability on isolated epidermal tissue. The epidermal sheets were placed in 96-well plates, each containing 120. mu.L of 5mg/mL MTT solution, and incubated at 37 ℃ for 1 hour. The resulting precipitate (purple formazan)Crystals) were dissolved in 120. mu.L of 2-propanol at room temperature for 15 minutes. Then, the absorbance of the staining solution was measured at 560nm (Multiskan EX, Thermo Scientific) using a microplate reader.

Lipid peroxidation end product

UVB-induced lipid peroxidation in the epidermal layer was determined using fluorometric quantitation of Malondialdehyde (MDA) levels (Oxiselect TMTBARS Assay Kit; Cell Biolabs, Inc.; catalog number: STA-330). The skin pieces were homogenized and the thiobarbituric acid reactant (TBARS) levels of the samples were determined directly according to the supplier's instructions. The final fluorescent product was examined in 96-well 96F Nunclon Delta Black Microwell SI (Thermo Scientific; Cat. No.: 137101) using a microplate reader Tecan Infinite M200 PRO.

LPS-induced pro-inflammatory HSOC mimetics

For HSOC, the strain was cultured in medium (Life Technologies Ltd.,the directory number: 41965-; catalog No. sc-3535) and Epithelial Growth Factor (EGF), mimicking the state of inflammation ex vivo; the LPS-containing solution was added systematically to the medium. After obtaining the inflammatory state, WPE or WSE was topically administered to HSOC without reconstitution of the culture medium and co-incubated for 48 hours. Two days after topical WPE or WSE, the media was collected and analyzed as described below.

Quantification of inflammatory cytokines in HSOC following LPS-induced stimulation

The effect of WPE or WSE on the inflammatory process was determined by measuring the levels of inflammatory cytokines (i.e., IL-6, IL-8, and TNF α) in the media of HSOC previously incubated with WPE or WSE. Specifically, collected DMEM samples for determination of IL-6 and IL-8 were cultured in fresh medium at a ratio of 1: 500 dilution and transfer to plate, while the collected DMEM samples for TNF α determination were transferred to plate without dilution. Enzyme-linked immunosorbent assay (ELISA) by sandwich method (Max) according to the manufacturer's protocol^TM Set Deluxe，San diego, california, usa) to determine the amount of a particular cytokine in each sample. The resulting products were measured quantitatively by Optical Density (OD) scanning at 450nm using a Tecan Infinite M200 PRO microplate reader.

Determination of radical scavenging Activity in 2, 2-Diphenyl-1-picrylhydrazino (DPPH) assay

2, 2-diphenyl(1-picrylhydrazino (DPPH) assayThe directory number: d9132) Are used to monitor chemical reactions involving free radicals. A working solution of 100 μ M DPPH was prepared; add 20 μ L of WPE or WSE to 380 μ L of DPPH solution in a 24-well plate; incubating for 30 minutes at room temperature in the dark to generate free radicals; 100 μ L of the reaction solution was transferred to a 96-well plate, and the absorbance at 517nm of each sample was measured on a microplate reader Tecan Infinite M200 PRO. The scavenging effect of the sample solution on DPPH radical solution was measured and converted into Trolox Equivalent Antioxidant Capacity (TEAC), by which the results were shown.

Electrospray ionization time of time-of-flight mass spectrometry (ESI-TOF-MS)

The samples were mass-analyzed using positive ESI-TOF-MS at constant flow rate. Data interpretation was performed by MassHunter profiler software.

Data analysis

Data are presented as mean ± Standard Error of Mean (SEM). Statistical analysis was performed using a two-tailed t-test. P values of less than 0.05(P <0.05) were considered significantly different compared to the untreated control group.

Example 1

Comparison of VOCs in WPE and WSE

The inventors characterized VOCs in WPE and WSE using gas chromatography. Some peaks were highly prominent and shared between WPE and WSE (fig. 1A-1B). The comparison of VOCs in WPE and WSE is summarized below (table). WPE was found to contain about 2.5 times more VOCs than WSE. Specifically, VOCs referred to as unknown 3 and unknown 15 were found to be more prominent in WPE than WSE, while VOCs referred to as unknown 10 were found in comparable amounts in both WPE and WSE (fig. 1A-1B).

TABLE-comparison of VOCs in WPE and WSE (VOCs of 0.5% w/w or lower are not present)

Example 2 comparison of antioxidant Activity and Total phenol content in WPE and WSE

The inventors quantified antioxidant activity and total phenol content in WPE and WSE. It was found that both WPE and WSE comprise phenolic compounds, wherein WPE comprises approximately 2 times more than WSE (figure 2). WPE has free radical scavenging activity in DPPH assay with antioxidant capacity of 6.7mg/Trolox equivalent/g dry weight. No antioxidant capacity of WSE was detected.

Example 3

WPE and WSE attenuate UVB-induced apoptosis in HSOC

The inventors found that the chemical composition of the plant, magadipine, changed over a relatively short period of time, from 7 months to 8 months. In fact, at early 7 months, WPE and WSE had a photoprotective potential, but still far from their maximum (fig. 3A). Furthermore, during this time, WPE and WSE were found to be highly toxic to HSOC (fig. 3B). However, by 7 months, the major changes begin to occur-the ability to prevent photodamage increases, and at the same time the cytotoxicity of the material decreases. This trend continues and by the beginning of 8 months, the emulsion reaches its maximum activity to protect the HSOC from harmful UV radiation. It is important to emphasize that during this period the cytotoxicity of the emulsion, if not totally eliminated, has reached an acceptable limit.

The inventors concluded that, between the upper and lower half months of 7 months, an important process occurred in the plants that allowed for the enhancement of the photoprotective components and the reduction of toxicity (fig. 3C). Furthermore, with respect to WPE, its effect was demonstrated to be dose-dependent (fig. 3D).

The inventors then showed that the final maturation and the achievement of the highest degree of biological activity of the plants during the 7 to 8 months is most critical. In this regard, with respect to the composition of WPE, the inventors showed that the main reservoir of active substances responsible for photoprotective activity and cytotoxicity is in the bark of the plant, not in its pith (fig. 3E).

Example 4

The light protection capability of the grease of the malt and balsam can be maintained under the condition of multiple times of irradiation

Successive tests of WPE and WSE performance of mecahuma showed impressive results in protecting HSOCs from harmful UV radiation (fig. 4A-4B). The HSOC was pretreated with WSE, WPE and WSE/WPE mixture and irradiated after incubation. The next day the samples were subjected to repeated irradiation. The uniqueness of this experiment is that it consists of two groups: (i) after the first irradiation (standard treatment); (ii) after two exposures. Evaluation of the photoprotective activity of the extracts showed that both WPE and WSE had a higher efficacy in preventing photodamage incurred by repeated irradiation.

Example 5

The oils WPE and WSE proved to be advantageous over other ether-based oils available (ethereal myrrha oil)

Disclosed are WPE and WSE with available myrrh essential oil widely used in the cosmetic industry; a commercial oil; oils produced by Elsinobia; and comparative analysis of raw oil from also (FIGS. 5A-5B). The results demonstrate that the emulsified extract produced in the Ein-Gedi area (during 2016. 8 months) is clearly advantageous over other extracts. For example, diluted commercial oils are less reactive and lose their photoprotective ability rapidly (FIGS. 5A-5B).

Example 6

The Gnetum melitensis WPE and WSE demonstrated their advantages over commercial sunscreens

In addition, the inventors performed comparative analyses of creams containing zinc oxide (ZnO), widely used cosmetic procedures and sunscreens as UV absorbers, or WPE and WSE prepared at 8 months (fig. 6A-6B). It is noteworthy that zinc oxide has lower photoprotective activity and greater cytotoxicity than organic creams containing WPE or WSE. Similar results were observed in the following experiments which tested the photoprotection and toxicity of popular mineral creams with different SPFs (from 15 to 100) (fig. 6C-6D). It was confirmed that the commercial mineral cream indeed has a strong photoprotective power and at the same time has a cytotoxic effect as evidenced by an increase in mitochondrial activity. In contrast, WPE (harvested at 8 months) had the greatest photoprotective activity with reduced toxicity.

Example 7

The melagatran WPE and WSE have synergistic photoprotective effects when added to sunscreen emulsions

The inventors then wanted to test whether there was a synergistic effect by combining the mugatran WPE with a synthetic sunscreen. The combination of the lowest concentrations of these two ingredients resulted in the greatest protective effect and balanced vigor when WPE was added to the sunscreen emulsion (fig. 7A-7B). In the following experiments, the possibility of synergistic interaction in preventing pyrimidine dimer (CPD) formation was also tested. The inventors clearly demonstrated that the use of a combination of sunscreen emulsion and WPE, WSE or various mixtures of both, enhanced the protective effect under the action of uv light and prevented the occurrence of mutagenic DNA defects compared to the effect induced by each of them when they were used alone (fig. 7C).

Example 8

WPE and WSE with dual antiinflammatory effects

The expression of pro-inflammatory cytokines was examined after induction of inflammation in HSOC by LPS and EGF and exposure to WPE and WSE (fig. 8A-C). WPE and WSE resulted in only a slight decrease in IL-6 production (FIG. 8A); in turn, the expression of IL-8 was virtually unaffected (FIG. 8B). In contrast, TNF α synthesis was significantly reduced (fig. 8C). The inventors also showed that WPE and WSE induced activation of the expression of the enzyme heme oxygenase-1 (HO-1), known to have cryoprotective properties (FIG. 8D).

While certain features of the invention have been described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.