BR102015027818A2 - cosmeceutical and / or dermatological composition, use of dermatological water composition and production process - Google Patents

Abstract

cosmeceutical and / or dermatological composition, use of dermatological water composition and production process. The following invention describes dermatological water composition composed of differentiated waters of natural extracts, conditioning agents and standardized minerals for skin hydration with low potential irritant action and epidermis regeneration after dermatological procedures, stimulating the synthesis of filaggrin and its natural factors. hydration. The present invention is in the field of pharmacy, medicine and chemistry.

Description

COSMETIC AND / OR DERMATOLOGICAL COMPOSITION, USE OF DERMATOLOGICAL WATER COMPOSITION AND PRODUCTION PROCESS Field of the Invention [1] The present invention describes cosmeceutical and / or dermatological compositions comprising dermatological waters composed of standardized natural extracts, conditioning agents and minerals. The product is preferably used for skin hydration, especially those that require compositions with low potential for irritant action and which additionally promote epidermis regeneration after dermatological procedures, stimulating the synthesis of filaggrin and its natural hydration factors. The present invention is in the field of pharmacy, medicine and chemistry.

Background of the Invention [2] Human skin is a vital organ that surrounds the body delimiting its contact with the external environment, performing the functions of thermal regulation, sensory functions such as heat, cold, pressure, pain and touch, organic defense, control. blood flow and protecting the body from excessive water loss.

[3] The skin has a structure with two distinct layers, the epidermis and the dermis. The epidermis is the outermost layer, being formed by epithelial tissue and presenting five layers, stratum corneum, lucid stratum, granular stratum, spinous stratum and germinal stratum.

[4] Epidermal differentiation begins with the migration of basal layer keratinocytes and ends with stratum corneum formation, with sequential proliferation, differentiation, and programmed cell death occurring, each of which is characterized by the action of specific proteins.

[5] The stratum corneum, the outermost layer of the epidermis, is highly organized and consists of keratin-rich dead cells and is formed and continuously regenerated by keratinocytes during the keratinization process.

[6] Acting as a protective barrier to underlying tissues against injury and infection, it also has a function of retarding evaporation, contributing to the maintenance of moisture in the deeper layers of the epidermis and the smooth, supple texture of the skin.

[7] The major protein acting in the formation of the stratum corneum is filaggrin. During the differentiation process, keratinocytes acquire keratohyaline granules consisting of profilagrin, the precursor protein of filaggrin. Prophylagrin is a complex, highly phosphorylated inactive polypeptide that undergoes protease dephosphorylation and cleavage releasing copies of functional filaggrin peptides.

[8] Filaggrin is so called because it has the function of aggregating keratin filaments into compacted bundles, giving the flattened aspect of the stratum corneum. There is also the production of other insoluble proteins and lipids that fill the intercellular spaces, contributing to the formation of an intact barrier.

[9] Over time filaggrin is progressively degraded by enzymes in urocanic acid and a mixture of hydrophilic amino acids that constitute the natural hydration factor (FHN), responsible for retarding water evaporation and essential for maintaining hydration of this layer. giving the skin youthfulness.

[10] Changes in the skin's natural hydration mechanism are unavoidable, usually linked to aging, sun exposure, pollution and eating and, as a result, cause feelings of discomfort, stretching, loss of silkiness, shine and softness, presence or increase of wrinkles, peeling, loss of elasticity and sensitivity.

[11] Still during the aging process, the skin undergoes several changes related to functional decline and clinically becomes thinner and less elastic. The amount of collagen and elastin, dermal fibers that give firmness and elasticity, are reduced and sagging with contour loss is frequent, both due to gravity and internal and external factors such as sun exposure, stress and genetic predisposition. . In addition to sagging as a result of sun exposure, surface spots may become more evident. To prevent or reduce this aging skin several medical and dermatological procedures are currently employed and belong to the field of cosmiatry, branch of dermatology mainly dedicated to human beauty and responsible for aesthetic skin care.

[12] Therefore, within cosmiatry, the goal of facial rejuvenation is to look younger by helping to reverse sun damage and aged appearance by reducing wrinkles, sun spots, sagging skin, improving texture and color. of the skin.

[13] Major advances in minimally invasive procedures and product and equipment technology enable dermatologists to achieve effective results. Procedures are less aggressive and safer, with faster recovery.

[14] To improve skin texture and stimulate fibroblasts, cells responsible for the production of dermal fibers, procedures such as intense pulsed light and peeling have been adopted for many years.

[15] According to the proposed treatment, we may observe some temporary side effects, such as mild redness, swelling, and some minor discomfort as a result of burning, itching or pinching. Scarring, persistent redness or permanent pigment change are not common.

[16] In this context, topical products for post-procedure use should be targeted to reduce discomfort and induce faster and more effective repair, reducing the risk of side effects.

[17] Thermal waters, for example, have a high concentration of mineral salts and trace elements that give them therapeutic properties, such as soothing and moisturizing, and are commonly derived from underground heating. Its dermatological use is routinely indicated for use after peeling and pulsed light procedures. However, as it is a natural compound, there is little control over the types and quantities of minerals present in this type of water.

[18] Several products are available on the market and waters are preferably classified according to their composition and origin, and for each characteristic there is an indication of a skin problem.

[19] Rusty waters are used for allergies and juvenile acne, Sodium chlorides relieve irritated skin, Sulphurous waters are used for skin diseases, Sulfated as anti-inflammatory, Calcic for eczema and dermatosis, Silicated as anti-inflammatory. Inflammatory and soothing, alkaline moisturizes the skin, acidic acid regulates skin pH, and oligominerals sanitize skin.

[20] Thus, a composition that, for example, has an intermediate pH, between alkaline and acid, would have the advantage of having these two characteristics of thermal waters, bringing skin hydration and regularization, especially by simulating the same pH commonly. found on "normal" skin.

[21] However, mineral salts when used in isolation do not penetrate the skin properly, having only a superficial action and acting primarily as a moisturizer for their ability to retain water. Until now there is some difficulty in standardizing these minerals, as their quantities and types depend on the characteristics of natural sources and may change over time. Thus, the possibility arises of binding dermal penetration peptides with said minerals, so that they penetrate the skin more appropriately.

[22] It is known that some natural extracts can also be used as skin conditioning antioxidants. Lotus root extract is marketed for use in cosmetic preparations, which in addition to bringing an antioxidant potential provides a soothing activity to the skin tissue.

[23] US 2008/199489 describes aqueous topical formulations for skin treatment comprising extracts of teas, algae, herbs, flowers, fruits and vegetables. Lotus, apple and grape extracts are included among the possible components of the formulations. They are topical formulations for the treatment of cancer, precancer, acne, rosacea, eczema, psoriasis, melasma, couperose and general inflammatory conditions. Such compositions use ozone and hydrogen peroxide to maintain the effectiveness of the formulation over time, and the treatment is complex in several steps.

[24] US 2005/281900 describes cosmetic compositions containing vinyl vitis extract used for NO synthase inhibition and treatment of NO-mediated disorders such as associated inflammatory neurogenic processes (rosacea) and proliferative disorders (psoriasis).

[25] US 6,468,564 describes compositions containing lotus seed extract for wrinkle reduction and expression lines. The use of lotus root extract in this document is not mentioned.

[26] Document CN 103405370 teaches preparations in the form of fruit water comprising mainly water mixed with apple, lemon, grape, beta-glucan water and other ingredients for hydration, instant repair, cleansing and skin refreshment.

[27] Surprisingly, the present invention finds that the combination of conditioning extracts with standardized minerals and additional conditioning agents in cosmeceutical and / or dermatological compositions, in addition to conditioning the skin, promotes filaggrin synthesis, accelerating stratum corneum restoration.

[28] The above state of the art documents use some extracts in cosmeceutical compositions to condition the skin and potentially alleviate irritation symptoms caused by various causes. However, none of these documents show any influence on the mechanism of stratum corneum formation promoting the restoration of the protective barrier and the natural maintenance of skin hydration, nor do they additionally provide standardized minerals in their composition in optimal amounts to aid [29]. They are achieved accordingly by the use of a dermatological water composition that combines the benefits of minerals and natural extracts, as well as cosmetically accepted conditioning excipients. The dermatological waters of the present invention are initially demineralized and artificially minerals are added in specific amounts to aid in its function.

[30] From what is clear from the researched literature, no documents were found anticipating or suggesting the teachings of the present invention, so that the technical solution proposed here has novelty and inventive activity in relation to the state of the art.

Summary of the Invention [31] It is an object of the present invention to provide a dermatological water composition containing natural differentiated waters for preferred use in dermatological post-procedures. Additionally, it is an object of the present invention to provide a composition comprising simultaneously additional conditioning agents with low skin irritation potential and standard minerals specially selected to enhance the beneficial properties of said composition.

[32] It is therefore an object of the present invention to provide a dermatological cosmeceutical and / or water composition comprising: a. at least one cosmetic grade water enriched with plant extract; B. at least one cosmetically acceptable conditioning agent; ç. at least one standardized mineral.

[33] In a preferred embodiment the water of different cosmetic grade comprises extracts obtained from flowers, fruits or roots. Even more preferably, the water is enriched with grape, apple and / or lotus extracts, among others, as well as the combination thereof.

[34] In a preferred embodiment, the conditioning agents comprise oligosaccharides, polysaccharides, among others, as well as a combination thereof.

[35] In a preferred embodiment, the standardized minerals are selected from the group comprising magnesium (Mg), zinc (Zn), iron (Fe), copper (Cu) and silicon (Si).

[36] In a preferred embodiment, minerals are biomineral. (and are covalently linked to peptides by a biotechnological process where free ions are not present, forming a complex commercially called Biominerals 5 (Highchem).

[37] In a preferred embodiment, the present invention comprises additional ingredients.

[38] The present invention comprises the use of a dermatological water composition comprising at least one plant extract, at least one cosmetically acceptable conditioning agent and at least one standardized mineral for the production of cosmetic and / or dermatological compositions or formulations.

[39] In a preferred embodiment, the composition of the present invention is used in dermatological post-procedures.

Brief Description of the Figures Figure 1 - Effects of the Dermatological Water product on PGE2 synthesis in human keratinocyte cultures.

Figure 2 - Effects of Dermatological Water on cell viability of human keratinocytes after 48 hours of incubation.

Figure 3 - Anti-Filaggrin immunofluorescence (green) and counterstaining with DAPI (blue; cell nucleus marker). 10 μm histological sections in ex vivo human skin fragments incubated with culture medium (images A, B, 400x) and skin fragments incubated with Project Lily product (images C and D, 400x) for 48 hours.

Figure 4 - Graph showing the percentage of improvement for the items: hydration and redness after product application (TImed), after 1.2 hours of single application and after 7 days of product use.

Figure 5 - Graph showing the percentage of improvement for the items: hydration, redness, feeling of quick comfort and soothing action after application of the product (TImed), after 1, 2 hours of single application and after 7 days of product use.

Figure 6 - Graph showing the percentage of improvement for the items: hydration and redness after product application (TImed), after 1.2 hours of single application and after 7 days of product use.

Figure 7 - Graph showing the percentage of improvement for the items: hydration, redness, sensation of rapid comfort and soothing action after application of the product (TImed), after 1, 2 hours of single application and after 7 days of product use.

Detailed Description of the Invention [40] The examples shown herein are intended solely to exemplify one of the numerous ways of carrying out the invention, but without limiting the scope thereof.

Dermatological Water Composition a. According to the invention, the dermatological waters of the present invention have conditioning action on the skin, providing the necessary hydration after dermatological procedures and additionally acting without causing irritating action on the skin and contributing to skin regeneration, which may mainly seen by increased synthesis of filaggrin. The dermatological water composition comprises at least one cosmetic grade water enriched with plant extract; at least one cosmetically acceptable conditioning agent; at least one standardized mineral. Cosmetic grade water with plant extract [41] The differentiated waters suitable for the present invention are deionized waters or reverse osmosis (and therefore cosmetic) waters mixed with natural extracts. In a preferred embodiment, the extract is obtained from flowers, fruits, roots, stem, leaves. Even more preferably, it is obtained from flowers, fruits and roots. Among the natural extracts, preferably, are the grape extracts (vitis vinifera), preferably the fruit, apple extract (pyrus malus), preferably the fruit, and lotus extract (nelumbo nucifera), preferably from the root and mixtures thereof. In a preferred embodiment, grape fruit extract and / or apple fruit extract and / or lotus root extract are obtained. In a preferred embodiment, between 0.01% and 10% of the extracts are mixed with cosmetic grade water, making it different from the state of the art. In an even more preferred embodiment, between 1% and 5% of the extracts obtained from fruits and / or flowers are mixed with cosmetic grade water. In an even more preferred embodiment, between 0.01% and 3% of the root extracts are mixed with cosmetic grade water. In a preferred embodiment, the composition comprises a minimum cosmetic grade water level of 80%. Standardized Minerals [42] Standardized minerals are preferably chosen from the group comprising Fe, Zn, Si, Cu, Mg. In a preferred embodiment, the minerals are biominerals. Biinerals of the present invention comprise organo-mineral bioactive, those minerals that are covalently linked to peptides by a biotechnological process where free ions are not present. These minerals are present in the complex in the same proportions found in plasma and because they are covalently bound to peptides can be rapidly absorbed by the skin, which does not occur in inorganic form. For each mineral there is an ideal amount of complexed protein that allows it to penetrate the skin optimally. [43] In a preferred embodiment, the biomerals of the present invention form a complex commercially called Biominerals 5 ® (Highchem).

Cosmetically acceptable conditioning agent [44] The cosmetically acceptable conditioning agents of the present invention comprise oligosaccharides, polysaccharides, among others, as well as the mixture thereof. In a preferred embodiment, other additional agents may be incorporated alone or in conjunction with the present invention, including humectants, additional plant extracts and fragrances and vitamins.

[45] Preferred polysaccharides according to the invention are, for example, gums. A particularly preferred representative is gum biosaccharides 1,2, 3, 4, fucose, galactose and galacturonic acid. In the present invention, a preferred polysaccharide is gum-1 biosaccharide in a mixture further comprising fucose, galactose and galacturonic acid. Particularly preferred representative here is Solabia's Fucogel® commercial product.

[46] Preferred oligosaccharides according to the present invention are, for example, alpha-glucan oligosaccharides and beta-glucan oligosaccharides. A particularly preferred representative is the oligosaccharide obtained by the fermentation of two natural sugars (sucrose and maltose), the preferred commercial product being Solabia Bioecolia®.

[47] Humectants are selected from the group comprising polyols, such as straight and branched chain alkyl polyhydroxyl compounds. For example, propylene glycol, sorbitol and glycerin are preferred. Polymeric polyols such as polypropylene glycol and polyethylene glycol are also useful. Butylene and propylene glycol are also preferred as penetration enhancers.

[48] Still from the group of preferred humectants is selected pentylene glycol which in addition to the moisturizing and conditioning properties of the skin has the secondary role of solvent and preservative of the formulation.

[49] Plant extracts and fragrances and aromas can also be used. According to the invention, fragrances and aromas comprise aromatic oils (fragrance) as well as aroma substances (odor). These are odorants, specifically fragrances. Fragrance base substances are generally essential oils, flower oils, plant extracts and animal drugs, natural product isolated odorants, chemically modified (semi-synthetic) odorants, and purely synthetic odorants. According to the invention, fragrances also include flavorings.

[50] Extracts, fragrances and aromas may originate here from numerous plant starting materials. Examples that may be specified are: flowers, for example lavender, rose, jasmine, arnica, neroli; trunks and leaves, for example of geranium, patchouli, petit-grain, fruits such as anise, coriander, caraway, juniper; fruit peels, for example from fruit, such as bergamot, lemon, orange; seeds such as mace, angelica, celery, cardamom, roots such as angelica, costus, iris, calmus; wood such as sandalwood, guaiac wood, cedar wood, rosewood; herbs and grasses such as tarragon, lemon grass, sage, thyme; needles and branches, for example, spruce, pine, pine, dwarf pine; resins and balms, eg galbanum, elemi, benzoin, myrrh, frankincense, opopanace.

[51] Illustrative vitamins of the present invention include vitamin A (retinol), vitamin B2, vitamin B3 (niacinamide), vitamin B6, vitamin C, vitamin E and biotin. Vitamin derivatives may also be employed. For example, vitamin C derivatives include ascorbyl tetraisopalmitate, ascorbyl magnesium phosphate and ascorbyl glycoside. Vitamin E derivatives include tocopheryl acetate, tocopheryl palmitate and tocopheryl linoleate. In a preferred embodiment, panthenol and its derivatives (D-panthenol, DL-panthenol) may also be employed.

[52] According to one embodiment of the invention, cosmeceutical and / or dermatological compositions are dermatological waters which may preferably be used in sprays and / or aerosols.

Example 1 - Cosmeceutical and / or dermatological composition [53] The specific but not limiting example is outlined below further illustrating the present invention.

Table 1 - Formulation Example according to the present invention.

Table 2 - Formulation Example according to the present invention.

Table 3 - Formulation Example according to the present invention. Example 2 - Composition Production Process [54] The production process of the cosmeceutical and / or dermatological composition is exemplified by the following steps: Add: a. at least one cosmetic grade water; B. at least one with plant extract; ç. at least one cosmetically acceptable conditioning agent; d. at least one standardized mineral. in a reactor with agitator and disperser for a suitable time to allow mixing thereof.

[55] The reactor is preferably an RPM anchor reactor with a decentralized Hz stirrer and RPM disperser.

[56] The stirring process is preferably carried out between 10 and 30 minutes, even more preferably for about 20 minutes.

[57] After obtaining the product, it is subjected to an in-process control analysis to verify physicochemical specifications of the composition.

Table 4 - Project-Lily Production Control Analysis Specification [58] The final product obtained is subjected to a 100 mesh screen. In a preferred embodiment, the product is packaged in 200 kg bags for transportation.

[59] The product obtained is packaged in personal care packaging, preferably packaging in which there is no user contact with the stored product.

[60] Most preferably the final product is packaged using bag-on-valve packaging.

Example 3 - Physicochemical Evaluation [61] The formulation prepared in Example 2 was analyzed after preparation, stored at room temperature and retested after 30 days, 60 days and 90 days. Results are presented in table 5.

Table 5 - Result of physicochemical analysis on T0 (after preparation), t30 (after 30 days storage at room temperature), T60 (after 60 days storage at room temperature) and T90 (after 90 days storage at room temperature) ) [62] The samples showed no physicochemical change during the storage period at room temperature.

Example 4- In vitro Evaluation of Anti-Inflammatory Activity [63] The study consists of the evaluation of the anti-inflammatory potential of cosmetic ingredients. Inflammatory mediators are chemicals secreted by various cell types that act to trigger or ameliorate specific aspects of inflammation. These mediators may be called eicosanoids or arachidonic acid (AA) metabolites and are derived from the phospholipid components of the cytoplasmic membrane.

[64] Prostaglandin E2 is an eicosanoid derived from AA. Intradermal injection of prostaglandin E2 (PGE2) causes vasodilation with increased blood flow and its concomitant administration with leukotrienes, in this case LTB4, results in amplification of inflammatory response, probably due to the direct effect of PGE2 on blood vessels.

[65] Regulation of the immune and inflammatory response is not restricted to immunocompetent cells only. Keratinocytes, melanocytes, fibroblasts and endothelial cells, located between the epidermis, dermis and hypodermis layers, are involved in a dynamic interaction capable of detecting a variety of disturbances in the skin environment and rapidly transmitting appropriate signals that alert and recruit components of the immune system.

[66] Keratinocytes comprise approximately 95% of the epidermal cell population and are primarily responsible for its structural integrity through the production of the horny layer and maintenance of the skin barrier through lipid synthesis. Once stimulated, these cells are able to release various factors and promote expression of a variety of receptors that are significantly involved in eicosanoid immunoregulation and biosynthesis.

[67] These factors may act autocrine or paracrine by influencing the multiple resident cell types of the skin, particularly endothelial cells. Thus, active products and ingredients capable of inhibiting the synthesis of proinflammatory skin mediators have significant potential for use in cosmetic dermatology.

[68] The effects of the product described in Example 1- Dermatological Water on the synthesis of the inflammatory mediator prostaglandin 2 (PGE2) in culture of basal conditions and stimulated with bacterial lipopolysaccharide (LPS) and phorbol acetate myristate were evaluated. (PMA) in order to simulate the inflammatory response. The purpose of this evaluation was to measure a possible in vitro antiinflammatory activity of the test product.

[69] Human keratinocytes (Cascade Biologics, USA) were seeded into 75 cm 2 bottles (Corning Inc, USA), cultured and expanded in a humid greenhouse at 37 ° C in the presence of 5% CO2 using specific culture medium. At confluence, cells were seeded in 06-well plates (Nunc, USA) for subsequent incubation with the test product and evaluation of the proposed mediator.

[70] Cell cultures were incubated with 3 non-cytotoxic concentrations of the product, previously determined by MTT technique. The concentrations evaluated in this study were 0.78; 0.39 and 0.19% (w / v). The cells were kept in contact with the test product and LPS / PMA (Sigma, USA) for 48 hours for subsequent supernatant collection.

[71] PGE2 concentration was measured in keratinocyte culture supernatant by a competitive binding assay (Cayman Chemical, USA). Samples were added to the polyclonal antibody coated ELISA plate. Then monoclonal antibody was added and the plate was kept at 4 ° C overnight. The developer was added and incubated for a further 90 minutes. The plate was subjected to absorbance reading at 405 nm. Results were calculated based on the reference curve of known concentrations of the PGE2 metabolite.

[72] The effects of Dermatological Water on PGE2 synthesis in human keratinocyte cultures are shown in Figure 1.

[73] As can be seen, Dermatological Water, when applied to human keratinocyte cultures at the evaluated conditions and concentrations (0.78; 0.39 and 0.19% (w / v)), did not promote alteration over PGE2 synthesis compared to the LPS / PMA control group. Example 5 - Evaluation of Cytotoxic Potential [74] The study consists of quantitative colorimetric assessment of mitochondrial metabolism and cellular respiratory chain activity in response to direct contact with xenobiotics. Verification of cell viability by the use of the MTT vital dye (3- (4,5 dimethyl thiazole-2yl) -2,5 diphenyl tetrazolium bromide) is one of the most widely used and cited methodologies in the scientific literature to determine the cytotoxic potential of substances or pharmaceutical, chemical, nutritional compounds as well as environmental factors. This assay is based on the conversion of yellow tetrazolium bromide (MTT) to blue formazan by mitochondrial succinate dehydrogenase enzyme in viable cells.

[75] This study aimed to determine the possible in vitro cytotoxic potential of Dermatological Water by MTT (3- (4,5 dimethyl thiazole-2-yl) -2,5 diphenyl tetrazolium bromide cytotoxicity assay). ).

[76] Cryopreserved human keratinocytes (Clonetics, Cambrex / Lonza, USA) were seeded into 75 cm 2 bottles (Corning Inc, New York, NY), cultured and expanded to at least the fifth pass in a humid greenhouse at 37 ° C in presence of 5% CO2 using specific culture medium. At approximately 80-90% confluence, cells were trypsinized (Trypsin-EDTA solution), neutralized, centrifuged (220 xg) for 10 minutes, counted in a Neubauer chamber and seeded in 96-well plates (Nunc, USA) for later. incubation with the test product and assessment of cytotoxic potential.

[77] Cell viability was determined by a colorimetric method using MTT (3- (4,5-dimethyl thiazole-2yl) -2,5 diphenyl tetrazolium bromide) dye (Sigma Chemical, St. Louis, Mo.), which It converts from yellow tetrazolium bromide (MTT) to formazan blue by the mitochondrial succinate dehydrogenase enzyme in viable cells. Human keratinocytes were seeded at a density of 2x10 4 cells / well in 96-well plates (Nunc, Roskilde, DM).

[78] The Dermatological Water test product was dissolved in the culture medium and added to the plate at a serial dilution in the range of 0.0004 to 50% (w / v). The culture was incubated for a period of 48 hours and MTT was then added to the culture at a concentration of 5 mg / mL (50 µl / well) which was incubated for a further 4 hours. The absorbance of each well was determined at 570 nm in a microplate reader. Cell death rate was expressed as a percentage as follows: Formula 1: Cell death rate [79] The product classification for in vitro cytotoxic potential followed the scale described in Table 6.

Table 6 - Scale for classification of cytotoxic potential established intra-laboratory for assets and cosmetic products.

[80] The results can be seen in Figure 2.

[81] According to the results presented in Figure 2, and following the classification of Table 6, Dermatological Water product showed significant cytotoxicity in the concentration range of 50 to 12.5% (w / v), which were responsible for up to 93.13% reduction in cell viability. In the concentration range below 12.5 to 1.56% (w / v), the product has moderate cytotoxicity. From a concentration of 0.78 to a concentration of 0.0004% (w / v), the product presents unimportant or negligible cytotoxicity, maintaining cell viability close to or greater than 100% when compared to the control.

[82] The Dermatological Water test product is safe for application to cell cultures at concentrations below 0.78%, considering the concentration range tested from 0.78 to 0.0004% (w / v), which expresses negligible or minor cytotoxicity.

Example 6 - Dermatological Evaluation of Photoirritant and Photosensitizing Potential Topical [83] Any topical product to be clinically evaluated should have preclinical data that delineate its safety margin for both its indication and its use. These data should include the systemic safety and use in the target organ itself, in the case of skin or mucosa, related to the risk of irritation or allergy (sensitization) where the product will be used.

[84] Topical products, whether finished products or ingredients, require human clinical trials to ensure that consumers are as safe as possible with the lowest risk of product use.

[85] The assessment of the cosmetic product in humans is not intended to investigate the potential for risk but to confirm the safety of the finished product. Thus, from the preclinical information collected, there should be proof of safety for use by humans. Topical compatibility tests aim to prove the safety of products in human skin.

[86] The literature's topical safety-proof protocols endorsed consist of applications under maximized conditions, with application area and controlled amount applied. The expected result is the absence of adverse reactions.

[87] To complement irritation and allergy studies, phototests (photoirritation and photosensitization) are indicated according to formulation and mode of use as well as intentional sun exposure products. Since exposure to solar radiation may trigger or aggravate adverse reactions to topical products, knowing the behavior of the product on human skin stimulated with ultraviolet radiation, according to the product in question, is of fundamental importance for safety evidence. topical.

[88] This study aims to prove the absence of the photoirritant and photosensitizing potential of a Dermatological Water product applied to the skin when exposed to sunlight through Phototest Assay.

[89] The study period was from 13/04/2015 to 14/05/2015. During this period, the volunteer recruitment and test execution steps were performed.

[90] 28 volunteers of both sexes were included according to the inclusion and non-inclusion criteria for the study. Inclusion criteria were age between 18 and 70 years, skin of the whole test region, agreement to obey the trial procedures and attend the clinic on the days and times determined for medical evaluations and for application and reading of the sign, term signature. phototypes: II or III, according to the Fitzpatrick adapted scale described in table 7.

Table 7 - Adapted Scale - Fitzpatrick TB: Soleil et peau. J Med Esthet 1975; 2: 33034 .___________________________________________________________________________ [91] The criteria for non-inclusion were pregnancy or lactation, prediction of sea bathing, swimming pool or bath during the study, water sports, use of anti-inflammatory drugs 30 days or immunosuppressants up to 3 months before selection, diseases that cause immunity suppression, such as Diabetes, HIV, etc., photosensitizing drug use, history of sensitization or photosensitization to topical products, active skin conditions that may interfere with study results, history or activity of photodermatoses, history photoinduced skin cancer, presence of skin cancer precursor lesions such as melanocytic nevi and actinic keratoses, intense sun exposure in the area of the experiment, use of new drugs / cosmetics during the experiment, previous participation in a study with it pr test product, relevant clinical history or current evidence of alcohol or other drug abuse, known history or suspected intolerance to any ingredient of the products under study (test products or comparators), professionals directly involved in the conduct of this study.

[92] Participating subjects were identified according to the research center identification procedure and a study inclusion sequence number. The sponsoring company provided the researcher with samples for evaluation in packaging with standard labels. Prior to study initiation, samples were identified with standardized labels as follows: Table 8 - Investigational Product Identification.

[93] Patching on the back of the volunteer occurred after preparation and application of the investigational products and controls in the semi-occlusive dressing. This preparation and application were determined according to the characteristics of the investigational product as shown below.

[94] Non-returnable products are applied pure. Supplies are physiological solution, mineral oil, 100U insulin syringes, gauze, Micropore tape and 100% cellulose patching paper (semi-occlusive patches consisting of 1 cm diameter filter paper discs and tape). semipermeable Micropore®) [95] On the first day of the study, after reading and signing the Informed Consent Form (ICF), the study volunteers were clinically evaluated and selected according to the inclusion and non-inclusion criteria. Once the inclusion was confirmed, the volunteer's skin was cleaned with gauze pad and physiological solution where the patches were applied. The study consisted of two steps: Dermal Photoirritation and Dermal Photosensitization.

[96] In dermal photoirritation - primary phase, volunteer patches containing the study product and the following control areas were applied to the back of the volunteers: physiological solution patch, mineral oil patch, patch without patch. After 24 hours, the drapes were removed and the region was irradiated with UVA radiation emitting equipment at a dose of 5 J / cm2. After 48 hours of irradiation, the area was again evaluated and the phase completed according to the schedule described in table 9 below.

Table 9 - Schedule of the Dermal Photoirritation Study - Primary Phase.

[97] At this stage, absences were not tolerated and, if this occurred, this would lead to the exclusion of the volunteer. The readings were taken 15 to 30 minutes after the removal of the dressing, according to the reading scale recommended by the International Contact Dermatitis Research Group - ICDRG (Table 10). Table 10 - Adapted Scale - RYCROFT, R. J. G. et al. Dermatitis - cumulative effect was given continued application on the back of volunteers containing patches containing the study product and the control areas cited in the previous step. After 24 hours, the volunteers returned to remove the beds and irradiate the area with UVA at a dose of 5 J / cm2. The volunteers returned for further irradiation and application for two weeks, making a total of six irradiations, as shown in table 11.

Table 11 - Dermal Photoirritation Study Schedule - Accumulated Effect [99] After the last irradiation the volunteers were at an interval of two weeks and returned to the challenge phase. In this step two groups of dressings were applied in areas that were not previously occluded. After 24 hours the volunteers returned for UVA irradiation at a dose of 5 J / cm2 in one area, the other area was spared. In both areas, evaluations were performed 48 hours after irradiation. The assessment was performed according to the reading scale recommended by the International Contact Dermatitis ResearchGroup - ICDRG (Table 10) and followed the schedule described in Table 12.

Table 12 - Schedule of Dermal Photosensitization Study - Challenge Phase.

[100] After the last step, volunteers underwent a dermatological assessment to verify the integrity of the test area. During the study, the volunteers were instructed not to move or wet the patches, not to start using topical products during the evaluation period and not to expose themselves directly to the sun. The volunteer attended the Institute at the scheduled times for reevaluations, as well as reported the use of any medication or treatment during the study.

[101] 28 volunteers were evaluated and selected for the study. After the photoirritation study period, none of the 28 volunteers who completed this stage had a skin reaction. After the photosensitization study period, none of the 28 volunteers who completed this stage had a skin reaction.

Table 13-Results obtained in the Primary Dermal Photoirritation and Accumulated Effect evaluations.

[102] Under the conditions under which the product described above was evaluated and in the sample of volunteers studied, the data allow us to conclude that no photoirritation potential was observed and no photosensitization potential of the Dermatological Water product was observed.

Example 7 - Assessment of Primary Dermal Irritability, Accumulated Dermal Irritability and Dermal Sensitization [103] The objective of the study was to verify the absence of potential for irritation (primary and accumulated dermal irritability) and allergy (sensitization) of the investigational product.

[104] The study period was from 06/04/2015 to 14/05/2015, and during this period the volunteer recruitment and trial run steps were performed. Fifty-five volunteers of both sexes were included, according to the inclusion and non-inclusion criteria for the study.

[105] Inclusion criteria were age 18 to 70 years, phototypes: I, II, III, and IV (according to Fitzpatrick adapted scale - Table 7), skin of the full test region, agreement to follow test procedures, and attend to the clinic on the days and times determined for medical evaluations and for application and reading of the wards, understanding, agreement and signing of the free and informed consent form.

[106] Criteria for non-inclusion were pregnancy and lactation, prediction of bathing, swimming or bathing during the study, water sports, anti-inflammatory drug use 30 days and / or immunosuppressive for up to three months prior to the study. screening, diseases that cause immunity suppression, such as diabetes, HIV, etc., personal history of atopy, history of sensitization or irritation to topical products, active (local and / or disseminated) skin disorders that may interfere with study results. , use of new drugs during the experiment, skin reactivity, previous participation in Compatibility Assays up to 21 days prior to the start of this study, volunteers with known congenital or acquired immunodeficiency, relevant clinical history or current evidence of alcohol or other drug abuse, known history or suspected intolerance to any ingredient of the products under study those under test or comparators), professionals directly involved in this study.

[107] Participating subjects were identified according to the research center identification procedure and a study inclusion sequence number.

[108] The sponsoring company provided the researcher with samples for evaluation in packaging with standardized labels. Prior to study initiation, samples were identified with standardized labels as follows: Table 14: Investigational Product Identification.

[109] Patching on the back of the volunteer occurred after product preparation and application, investigational and controls, in the semi-occlusive dressing. This preparation and application were determined according to the characteristics of the investigational product, as shown below.

[110] Rinsable (rinse) products are applied diluted to 5% in physiological solution or mineral oil according to solubility. Supplies are physiological solution, mineral oil, 100U insulin syringes, gauze, Micropore tape and 100% cellulose patching paper (semi-occlusive patches consisting of 1 cm diameter filter paper discs and tape). semipermeable Micropore®) [111] On the first day of the study, after reading and signing the Informed Consent Form (ICF), the study volunteers were clinically evaluated and selected according to the inclusion and non-inclusion criteria. Once the inclusion was confirmed, the volunteer's skin was cleaned with gauze pad and physiological solution where the patches were applied. The study consisted of three steps: primary dermal irritability, accumulated dermal irritability and dermal sensitization [112] In the assessment of primary dermal irritability (PID), volunteer patches containing the test product and the following control areas were applied to the back of the volunteer: physiological, mineral oil dressing, product-free dressing and tape (area without Micropore® dressing).

[113] After two days (48 h), the beds were removed and the region was evaluated by the clinical research assistant. In cases of positive evidence, the assistant was instructed to read again after 30 minutes of rest and, in case of permanence of the indication, the volunteer would be referred to the doctor. A new evaluation was performed after 96h, completing the Primary Dermal Irritability step. In this stage, absences were not tolerated and, if they occurred, resulted in the exclusion of the volunteer. The evaluations were performed according to the reading scale recommended by the ICDRG (Table 10). Table 15- Schedule of the Primary Dermal Irritability Study.

[114] For the Accumulated Dermal Irritability (IDA) study, continuity of application on the back of volunteers after the study product and the control areas mentioned in the previous step was given. The volunteers returned for removal of the patches, site evaluation and patch application for three weeks, thus completing eight applications. At this stage, each visit had a window of ± 1 day. In case of absence, the volunteer was instructed to remove the adhesive tape from the back, discard it and return on the next scheduled date. In cases of positive evidence, the assistant was instructed to read again after 30 minutes of rest and, in case of permanence of the indication, the volunteer would be referred to the doctor.

Table 16 - Schedule of the Accumulated Dermal Irritability study.

[115] Following the last assessment described in the previous item, volunteers were on an interval for two weeks and returned to the sensitization (challenge) phase, in which patches, test product and controls were applied to areas that were not previously occluded. According to Table 17, the volunteers returned for the following evaluations (48 and 72 h). In cases of positive evidence, the assistant was instructed to read again after 30 minutes of rest and, in case of permanence of the indication, the volunteer would be referred to the doctor.

Table 17- Schedule of the Dermal Sensitization (SD) study.

[116] After the last step, the volunteers underwent a dermatological assessment to verify the integrity of the test area. During the study, the volunteers were instructed not to move or wet the patches, not to start using topical products during the evaluation period and not to expose themselves directly to the sun. The volunteer attended the Institute at the scheduled times for reevaluations, as well as reported the use of any medication or treatment during the study.

[117] 55 volunteers were selected, 50 volunteers included completed the study. Volunteers with inclusion numbers 13 and 25 gave up participating in the study for work reasons from D3. Volunteer 24 gave up participating in the study for reasons of travel from D3. Volunteers with inclusion number 29 and 35 did not return from D36, three unsuccessful telephone contacts were made.

Table 18- Results obtained in assessments of Primary Dermal Irritation (IDP) and Cumulative Dermal Irritation (IDA).

[118] After the Dermal Sensitization study period, none of the 50 volunteers who completed this stage had a skin reaction, as shown in Table 19.

Table 19-Results obtained in Dermal Sensitization (SD) assessments Table 20- Legend of sites and sites [119] Under the conditions in which the product described above was evaluated and in the sample of volunteers studied, the data allow us to conclude that no potential was observed. of Dermal Irritation, no potential of Accumulated Dermal Irritation, no potential of Dermal Sensitization was observed.

Example 8 - Ex vivo Evaluation of Skin Barrier Benefits [120] Skin barrier integrity and maintenance of water balance cannot be considered solely by the presence of specific substances, but by the complex homeostasis of their overall composition.

[121] The epithelium of the skin, the epidermis, is in a constant balance between growth and differentiation and has a remarkable ability to renew itself. This homeostasis is important for the maintenance of this cell renewal and depends on several factors, one of them being an adequate adhesion to the basement membrane. The organization of this membrane is the result of a joint action of fibroblasts and keratinocytes, which form a complex multi-molecular structure. Cell adhesion is mainly mediated by integrins, fibronectin and laminins that act by connecting the extracellular matrix to the cytoskeleton. In addition, laminin plays an important role in regulating tissue architecture, migration and cellular signaling.

[122] Another part of this complex equilibrium is managed by the epidermal cornified envelope (ECE), a lipid-protein layer that replaces the plasma membrane of corneocytes and consists of a complex mixture of covalently linked proteins associated with a lipid-characteristic layer attached to the surface. extracellular protein layer5. Many constituents of ECE have been identified, among them prophylagrin, filaggrin, claudin-1, involucrin, loricrin, several keratin subtypes, among others.

[123] In this study, the ability of the Dermatological Water product to promote skin barrier enhancement through increased filaggrin production in human skin fragments was evaluated. Fragments (ex vivo) of human tissue were purchased from elective surgeries (blepharoplasty), and the study began on 4/14/2015 and was completed on 5/12/2015.

[124] After the surgical procedure performed by the ophthalmic plastic surgery clinical staff, eyelid fragments were placed in vials containing 0.9% saline until the time of the experiment. Biological samples were taken from the saline solution, immersed in 70% ethanol for 15 seconds and washed twice in new saline solution. The fragments were transferred to a petri dish containing culture medium for up to 48 hours for treatment with Dermatological Water product. The product was applied to the skin at a rate of 2 mg / cm2.

[125] After being incubated with the test product, ex vivo skin fragments were fixed in 4% Paraformaldehyde (pH 7.4) for 24 hours and cryoprotected in 30% sucrose solution for 48 hours. Then, 10 μm serial sections were collected directly on silanized slides with the aid of Cryostat (Leica - CN1850).

[126] At the end of harvesting, they were washed with 0.1 M PB and incubated overnight with the primary Anti-Filaggrin antibody (Santa Cruz; sc-AKH1, sc-66192). They were then rewashed with 0.1 M PB and incubated for 1 hour with Goat anti-Mouse secondary antibody (Invitrogen; A11001). At the end of these steps, a fresh incubation (1 minute) was performed with DAPI (4'-6-Diamidine-2-Phenylindole; DNA marker) followed by 3 washes of 5 minutes with 0.1 M PB.

[127] The slides were then mounted in a specific mounting medium and analyzed under a Fluorescence Microscope (Leica - DM 1000) with the aid of LAS Software (Leica Application Suite). The evaluated parameter was the fluorescence intensity emitted by the specific antibody labeling.

[128] According to Figure 3, the intensity of the green-labeled Anti-Filaggrin signal in the stratum corneum of the Dermatological Water-treated skin fragment (images C and D) was significantly higher compared to the basal control group (images A and B) from 48 hours of incubation. This result demonstrates that the Dermatological Water product intensifies the production of filaggrin in the skin, which will result in the direct formation of MNFs (natural skin moisturizing factors), as well as assisting the cohesion of the stratum corneum, having effects on the function and skin barrier integrity and maintenance of skin water balance.

[129] According to the results presented, it is concluded that Dermatological Water has been shown to increase the synthesis of filaggrin in human skin fragments and has been shown to contribute to the maintenance of skin water balance, maintenance and skin barrier integrity. Example 9 - Evaluation of Effectiveness in Improving Moisturizing and Feeling of Skin Comfort after Dermatological Procedure (130) The purpose of the study was to evaluate the effectiveness in improving skin comfort hydration after dermatological procedure (peeling). The study period was from 12/05/2015 to 06/04/2015 and during this period the volunteer recruitment and test execution steps were performed.

[131] 26 female volunteers were included according to the inclusion and non-inclusion criteria for the study. Inclusion criteria were female volunteers aged 30 to 60 years, volunteers who already use sunscreen daily, clinical indication for the protocol procedures: superficial chemical exfoliation, agreement to obey the test procedures and attend the clinic on days. and times determined for applications and / or evaluations, signing the Informed Consent Form.

[132] Non-inclusion criteria were pregnancy / lactation, use of anti-inflammatory / immunosuppressive / antihistamine drugs, atopic or allergic history of topical products or components of the formulation, active (local and / or disseminated) skin disorders. in the area of evaluation, pathologies that cause immunity suppression, such as diabetes, HIV, etc., history of cold sores, intense sun exposure up to 15 days before evaluation, dermatological treatments up to 4 weeks before selection, other conditions considered by the researcher as reasonable for disqualifying study participation (should be described in observations in the medical record).

[133] Participating subjects were identified according to the research center identification procedure and a study inclusion sequence number.

[134] Prior to study initiation, samples were identified with standardized labels as follows: Table 21: Investigational Product Identification.

[135] The mode of use of the investigational product was to spray at any time of day on the skin and to be absorbed for about 2 to 3 minutes. The supplies used were Jessner's solution (14% lactic acid, 14% salicylic acid, 14% resorcinol, qsp alcohol), gauze.

[136] Volunteers were initially recruited from the criteria indicated in the SELECTION OF VOLUNTEERS. Only individuals who had the necessary requirements according to the selection criteria mentioned above, and who understood, accepted and signed the Informed Consent Form could be included in the study.

[137] Volunteers were initially evaluated by a dermatologist for inclusion and non-inclusion criteria.

[138] The procedure below was performed at the Clinical and Surgical Dermatology Service of the Medcin Evaluation Laboratory. The procedure was performed by dermatologists at a specific location for such procedures. A nurse or Clinical Research Assistant (APC) was available to assist procedures and guide volunteers.

[139] Superficial chemical exfoliation consists of applying a chemical capable of exfoliating the skin to the viable epidermis and is indicated for the treatment of mild to moderate photoaging. The exfoliating agent used was Jessner's Solution (14% lactic acid, 14% salicylic acid, 14% resorcinol, qsp alcohol), recognized in dermatological practice, applied to the entire face of the volunteer.

[140] The first application of the investigational product was carried out at the Institute by the Clinical Research Assistant after peeling. The application of the investigational product was performed in hemiface, following a previous randomization.

[141] Immediately after application of the investigational product, volunteers answered a subjective questionnaire (immediate T) and after 1 and 2 hours, the summary of which is given below for reference.

Clinical evaluation> Regarding the area not treated with the product, hydration: () There was no difference (0); () The treated area is slightly more hydrated (1); () Slightly more hydrated (2); () More hydrated (3); > Regarding the area not treated with the product, the redness: () There was no difference (0); () The treated area is slightly less red (1); () Slightly less red (2); () Less red (3).

Subjective assessment> (Hydration) Regarding the area not treated with the product, you could say that: () There was no difference (0); () The treated area is slightly more hydrated (1); () Slightly more hydrated (2); () More hydrated (3). > (Redness) Regarding the area not treated with product, you could say that: () There was no difference (0); () The treated area is slightly less red (1); () Slightly less red (2); () Less red (3). > After application, did you have the sensation of rapid comfort on your skin? () Yes () No> (Soothing Action) Did you feel a quick soothing (refreshing) sensation on the treated skin? () Made no difference (0); () Improved slightly (1); () Improved (2); () It has improved a lot (3).

[142] At the end of the initial evaluations, volunteers were given an SPF 50 sunscreen and investigational product, which were used for seven (7) days throughout the facial region. Sunscreen was provided by Medcin along with the investigational product (sunscreens were provided to Medcin by the study sponsor).

[143] As noted above, volunteers completed a subjective questionnaire (see paragraph 146) immediately after application, after 1, 2 hours of hemiface application, and after 7 days of use of the investigational product.

[144] Twenty-six volunteers were evaluated in the study. Of these 25 were included. The age range of volunteers included was 32 to 58, with a mean age of 45 years. One volunteer (inclusion number: 21) did not return for final evaluation due to personal reasons unrelated to the study.

[145] An adverse event occurred in the study, volunteer inclusion number 19 returned to the institute at the study's final visit on May 28, 2015 with the presence of a 1.0 cm hyperchromic macula in the supra labial region. According to the dermatologist, the reaction is of mild intensity, probably related to the exfoliation procedure and with a diagnostic hypothesis of post-inflammatory hyperchromy. The volunteer did not discontinue use of the product. The dermatologist has prescribed Klassis cream, which should be applied to the lesion at night.

[146] The subject was permanently discontinued from the study and was scheduled to return for follow-up on 06/11/2015. Volunteer return to the institute on 06/11/2015 to finish an adverse event. The event ceased on 10/06/2015 and the dermatologist doctor suspended the treatment. In an internal discussion with the clinical team, it was found that the stain came from a cosmiatric procedure. The event was resolved without sequelae and the participant was permanently discontinued from the study.

[147] Twenty-three volunteers completed the study and none showed reaction reported or reported at the evaluation site under dermatological control.

[148] Figure 4 graphs percentages of improvement obtained in clinical facial region assessments performed immediately after product application (T Imed), after 1, 2 hours of single application, and after 7 days of product use.

[149] Below is a table with the comparison test (Wilcoxon) of the experimental time in relation to the central scale of the parameters: hydration and redness.

Table 22: Comparison test (Wilcoxon) in relation to the central scale and response percentages for each grade in their respective times.

[150] It is noted that the investigational product promoted a statistically significant improvement (p <0.05) for hydration and redness at all experimental times.

[151] After 7 days of use of the investigational product, the volunteers underwent clinical safety assessments as per the results in table 23 below: Table 23- Response Percentages for each grade [152] It is noted that the investigational product was well tolerated and most volunteers showed no signs or feelings of discomfort.

[153] Subjective assessment is presented in Figure 5 with a graph showing the percentages of improvement obtained in subjective facial region assessments performed immediately after product application (TImed), after 1, 2 hours of single application and 7 days after application. product use.

[154] Below is a table with the comparison test (Wilcoxon) of the experimental time in relation to the central scale of the parameters: hydration, redness, sensation of rapid comfort and soothing action.

Table 24: Comparison test (Wilcoxon) in relation to the central scale and response percentages for each grade in their respective times.

[155] It is noted that the investigational product promoted an all-time improvement in the parameters hydration, redness, feeling of comfort and soothing action. A statistically significant improvement (p <0.05) was observed for the items hydration, redness and soothing action at all experimental times except at T1h for redness. Most volunteers (95.7%) positively evaluated the calming action of the product at all experimental times.

[156] Under the conditions under which the product described above was evaluated and in the sample of volunteers studied, the data allow to conclude that: [157] In the clinical evaluation the investigational product promoted a statistically significant improvement (p <0.05) of hydration and Redness of the skin compared to the control area at the times immediately after application of the product, after 1 and 2 hours and after 7 days of continued use was well tolerated and most volunteers showed no signs or feelings of discomfort.

[158] Subjective evaluation of the investigational product promoted an all-time improvement for hydration, redness, comfort, and soothing action, with a statistically significant improvement (p <0.05) for hydration and Redness after 2 hours of application of the product in relation to the Timed time and most volunteers (95.7%) positively evaluated the calming action of the product at all experimental times.

[159] The investigational product has shown a calming action after performing a superficial peeling session, promoting increased hydration, a reduced erythema and a comfortable feeling with a calming effect and may be recommended for application after superficial exfoliative procedures, reducing patient discomfort and promoting faster and more effective recovery.

Example 10 - Evaluation of Efficacy in Improving Moisturizing and Feeling of Skin Comfort after Dermatological Procedure (Intense Pulsed Light) [160] The objective of the study was to evaluate the effectiveness in improving skin hydration and feeling of comfort following a dermatological procedure.

[161] The study period was from 06/17/2015 to 07/08/2015 and during this period the volunteer recruitment and test execution steps were performed.

[162] 35 female volunteers were included according to the inclusion and non-inclusion criteria for the study. Inclusion criteria were female volunteers aged 30 to 60 years, volunteers who already use sunscreen daily, clinical indication for the protocol procedures: intense pulsed light, agreement to obey the test procedures and attend the clinic on days. and times determined for applications and / or evaluations, signing the Informed Consent Form.

[163] Non-inclusion criteria were pregnancy / lactation, use of anti-inflammatory / immunosuppressive / antihistamine drugs, atopic or allergic history of topical products or components of the formulation, active (local and / or disseminated) skin disorders in the area of evaluation, pathologies that cause immunity suppression, such as diabetes, HIV, etc., history of cold sores, intense sun exposure up to 15 days before evaluation, dermatological treatments up to 4 weeks before selection, other conditions considered by the researcher as reasonable for disqualifying study participation (should be described in observations in the medical record).

[164] Participating subjects were identified according to the research center identification procedure and a study inclusion sequence number.

[165] Prior to study initiation, samples were identified with standard labels as follows: Table 25: Investigational Product Identification.

[166] The mode of use of the investigational product was to spray on the skin at any time of day and allow it to be absorbed for about 2 to 3 minutes.

[167] Volunteers were initially recruited from the criteria indicated in the selection of volunteers. Only individuals who had the necessary requirements according to the selection criteria mentioned above, and who understood, accepted and signed the Informed Consent Form could be included in the study.

[168] Volunteers were initially evaluated by a dermatologist for inclusion and non-inclusion criteria.

[169] The procedure below was performed at the Clinical and Surgical Dermatology Service of the Medcin Evaluation Laboratory. The procedure was performed by dermatologists at a specific location for such procedures. A nurse or Clinical Research Assistant (APC) was available to assist procedures and guide volunteers.

[170] Intense pulsed light (LIP) or pulsed light (LP) is not exactly a LASER because it emits non-coherent light with a continuous wave spectrum of 500 to 1200nm, pulses ranging from 0.5 to 20msec in sequence. (single, double or triple), photons target melanin, hemoglobin and water contained in collagen. The device (Limelight) uses specific filters that separate visible light by selecting wavelength for the treatment of deeper or deeper lesions. Patients will undergo a Limelight session (one pass) with the power of 14J and pulse mode "B".

[171] The first application of the investigational product was performed at the Institute by the Clinical Research Assistant after the realization of pulsed intense light (LIP). The application of the investigational product was performed in hemiface, following a previous randomization.

[172] Immediately after application of the investigational product, volunteers answered a subjective questionnaire (Timediate) and after 1 and 2 hours (quoted above in paragraph 146).

[173] At the end of the initial evaluations, the volunteers were given an SPF 50 sunscreen and the investigational product, which should be used every day for seven (7) days throughout the facial region. Sunscreen was provided by Medcin along with the investigational product (sunscreens were provided to Medcin by the study sponsor).

[174] As noted earlier, volunteers answered a subjective questionnaire (see paragraph 146) shortly after application, after 1, 2 hours of hemiface application, and after 7 days of use of the investigational product.

[175] Immediately after application of the investigational product to hemiface (TImed) and after 1, 2 hours of product application and after 07 days, the evaluating physician evaluated the following parameters: hydration, redness and safety assessment.

[176] 35 volunteers were evaluated in the study. The age range of volunteers included was 30 to 58, with a mean age of 43 years. One volunteer (inclusion number 20) withdrew from participating in the study after signing the consent form due to personal reasons unrelated to the study.

[177] Ten volunteers (inclusion numbers: 04, 07, 09, 10, 12, 13, 25, 26, 28 and 31) were considered as selection failure due to the following reasons: [178] Volunteer inclusion numbers: 04, 07, 10, 12 and 13 - Did not meet the following inclusion criteria: "Clinical indication for protocol procedures: Intense Pulsed Light".

[179] Volunteer inclusion number: 09 - Did not meet the following inclusion criteria: "Clinical indication for protocol procedures: Intense Pulsed Light" and fulfills the non-inclusion criterion: "Active (local and / or disseminated) skin disorders in the evaluation area ”.

[180] Volunteer inclusion numbers: 25, 28 and 31 - Fulfilled the following non-inclusion criteria: "Other conditions considered by the researcher to be reasonable for disqualifying study participation (should be described in observations on the clinical form)."

[181] Volunteer inclusion number: 26 - Has met the following non-inclusion criteria: "Other conditions considered by the researcher to be unreasonable for disqualification from study participation (should be described in observations on clinical form)" and "Intense sun exposure up to 15 days before the evaluation ”.

[182] Two volunteers (inclusion numbers: 18 and 35) did not return for final evaluation due to personal reasons unrelated to the study.

[183] Twenty-two volunteers completed the study and none showed reaction reported or reported at the evaluation site under dermatological control.

[184] Figure 6 graphs percentages of improvement obtained in clinical facial region evaluations performed immediately after product application (TImed), after 1, 2 hours of single application, and after 7 days of product use.

[185] Below is a table with the comparison test (Wilcoxon) of the experimental time in relation to zero scale (0 = no difference) of the parameters: hydration and redness.

Table 26: Comparison test (Wilcoxon) in relation to zero scale (0 = no difference) and response percentages for each grade in their respective times.

[186] The investigational product promoted a statistically significant (p <0.05) improvement in skin hydration and redness compared with the control area at the time immediately after product application, after 1 and 2 hours. This improvement was also significant for the hydration item after 7 days of continued use.

[187] For the redness item after 7 days, the null scale was statistically higher than the other improvement items, because the erythema had already ceased in the initial evaluations and the clinician evaluated as no difference compared to the initial skin.

[188] After 7 days of use of the investigational product, volunteers underwent clinical safety assessments as follows from the results in the table below.

Table 27- Response percentages for each grade.

[189] The investigational product was found to be well tolerated and most volunteers showed no signs or feelings of discomfort.

[190] Figure 7 shows the graph with percentages of improvement obtained in subjective facial region assessments performed immediately after product application (TImed), after 1, 2 hours of single application, and after 7 days of product use.

[191] Below is a table with the comparison test (Wilcoxon and Z-test for sensation of rapid comfort) of the experimental time in relation to the zero scale (0 = no difference / 0 = no) of the parameters: hydration, redness, sensation of Quick comfort and soothing action.

Table 28: Comparison test (Wilcoxon and Z test) in relation to the null scale (0 = no difference / 0 = no) and response percentages for each grade in their respective times.

[192] The investigational product was observed to promote a statistically significant improvement (p <0.05) for the items hydration, redness, sensation of rapid comfort and soothing action at all experimental times.

[193] Under the conditions under which the product described above was evaluated and the sample of volunteers studied, the data allow to conclude that: [194] In the clinical evaluation the investigational product promoted a statistically significant improvement (p <0.05) of hydration and skin redness, compared to the control area, at times immediately after, after 1 and 2 hours of single application of the product. This improvement was also statistically significant (p <0.05) for the hydration item after 7 days of continued use and was well tolerated and most volunteers showed no signs or feelings of discomfort.

[195] Subjective evaluation found that the investigational product promoted a statistically significant improvement (p <0.05) for hydration, redness, a sense of rapid comfort, and soothing action at all experimental times.

[196] The investigational product demonstrated a calming action after performing a Limelight session - intense pulsed light (one stride, 14J power and pulse mode B), promoting increased hydration, reduced erythema, and a sensation of comfort, with a calming effect and may be recommended for application after intense pulsed light procedures, reducing patient discomfort and promoting faster and more effective recovery [197] Based on the results obtained from the studies presented, it can be concluded that the Product Lily besides promoting the desired hydration after dermatological procedure (peeling and pulsed light) relieving discomfort also stimulates the synthesis of filaggrin accelerating the regeneration of the epidermis.

[198] Those skilled in the art will enhance the knowledge presented herein and may reproduce the invention in the embodiments disclosed and in other embodiments within the scope of the appended claims.

Claims (26)

1. COSMETIC AND / OR DERMATOLOGICAL COMPOSITION of dermatological water characterized by comprising: a. at least one cosmetic grade water enriched with plant extract; B. at least one cosmetically acceptable conditioning agent; ç. at least one standardized mineral. Composition according to Claim 1, characterized in that it comprises enriched cosmetic grade water comprising extracts obtained from flowers, fruits and / or roots. Composition according to either of Claims 1 and 2, characterized in that the extracts of the enriched water are selected from the group comprising grape, apple and / or lotus extracts. Composition according to any one of Claims 1 to 3, characterized in that the cosmetic grade water is enriched with 0.01% to 10% of plant extract. Composition according to Claim 1, characterized in that the conditioning agents comprise oligosaccharides and / or polysaccharides. Composition according to Claim 5, characterized in that the polysaccharides comprise biosaccharides. Composition according to Claim 6, characterized in that the biosaccharides comprise biosaccharide gum-1, fucose, galactose, galacturonic acid or a mixture thereof. Composition according to Claim 5, characterized in that the oligosaccharides comprise alpha-glucan oligosaccharide and beta-glucan oligosaccharide. Composition according to Claim 1, characterized in that the standardized minerals are selected from the group comprising magnesium (Mg), zinc (Zn), iron (Fe), copper (Cu), silicon (Si) and the mixture. of the same. Composition according to either of Claims 1 and 9, characterized in that the minerals are biominerals. Composition according to any one of Claims 1, 9 or 10, characterized in that the biinerals are covalently linked to peptides by a biotechnological process where free ions are not present. Composition according to Claim 1, characterized in that it comprises additional ingredients. Composition according to Claim 12, characterized in that it further comprises humectants, vitamins, vitamin derivatives, extracts, fragrances, aromas as well as a combination thereof. Composition according to Claim 13, characterized in that the humectants are selected from the group comprising polyols, straight and branched chain alkyl polyhydroxyls and polymeric polyols. Composition according to Claim 14, characterized in that the straight and branched chain alkyl polyhydroxyl polyols are selected from the group comprising propylene glycol, sorbitol, glycerine, butylene glycol, pentylene glycol and a mixture thereof. Composition according to Claim 13, characterized in that the vitamins are selected from the group comprising vitamin A (retinol), vitamin B2, vitamin B3 (niacinamide), vitamin B6, vitamin C, vitamin E , biotin as well as the combination thereof. Composition according to Claim 13, characterized in that the vitamin derivatives are selected from the group comprising ascorbyl tetraisopalmitate, ascorbyl magnesium phosphate, ascorbyl glycoside, tocopheryl acetate, tocopheryl palmitate, tocopheryl linoleate, DL-panthenol and its derivatives and mixtures thereof. Composition according to Claim 13, characterized in that the extracts, fragrances and aromas are selected from the group comprising lavender flowers, rose flowers, jasmine flowers, arnica flowers, neroli flowers, trunks and geranium leaves. , patchouli trunks and leaves, petit-grain trunks and leaves, fruits such as anise, coriander, caraway, juniper, bergamot peel, lemon peel, orange peel, mace seed, angelica seed, celery seed, cardamom seeds, angelica root, costus root, iris root, calmus root, sandalwood, guaiac wood, cedar wood, rosewood, herbs and grasses such as tarragon, lemon grass, sage, thyme , spruce needles and branches, pine, pine, dwarf pine, resins and balms of galbanum, elemi, benzoin, myrrh, frankincense, opopanace. Composition according to Claim 1, characterized in that it contains at least 80% of cosmetic grade water. Use of a dermal water composition according to Claim 1, characterized in that it is for the production of cosmetic and / or dermatological compositions or formulations. Use according to claim 20, characterized in that it is used in dermatological post-procedures. Use according to claim 20, characterized in that it is used in minimally invasive dermatological post-procedures. 23. Production process of a cosmeceutical and / or dermatological composition comprising the step of adding: a. at least one cosmetic grade water; B. at least one with plant extract; ç. at least one cosmetically acceptable conditioning agent; d. at least one standardized mineral. in a reactor with agitator and disperser for a suitable time to allow mixing thereof. A process according to claim 23, characterized in that the reactor is an RPM anchor reactor and a decentralized Hz stirrer and RPM disperser. Process according to Claim 23, characterized in that it is carried out for about 20 minutes. Process according to Claim 23, characterized in that the final product is subjected to a 100 mesh screen.