Antioxidant cosmetic complex, cosmetic composition containing same, use of the antioxidant cosmetic and method for cosmetic treatment

Abstract

The present invention relates to an antioxidant cosmetic complex for acting against damage caused by oxidative stress and combating free radicals (ROS) caused by UV radiation and pollution in the hair strands and scalp skin, comprising active ingredients from Euterpe oleracea (açaí), Theobroma grandiflorum (cupuaçu), Theobrama cacao (cocoa) and recombinant silk protein, as well as a cosmetic composition containing same, use thereof and method.

Description

[0001] “ANTIOXIDANT COSMETIC COMPLEX, COSMETIC COMPOSITION CONTAINING THE SAME, USE OF THE ANTIOXIDANT COSMETIC AND METHOD FOR COSMETIC TREATMENT” FIELD OF THE INVENTION

[0002] [1] The present invention relates to an antioxidant cosmetic complex, to act against damage caused by oxidative stress and combat free radicals (ROS) caused by UV radiation and pollution, in hair strands and scalp skin, comprising active ingredients from Euterpe oleracea (açaí), Theobroma grandiflorum (cupuaçu), Theobrama cacao (cocoa) and recombinant silk protein, as well as a cosmetic composition containing the same, its use and method.

[0003] BACKGROUND OF THE INVENTION

[0004] [2] The body's cells produce free radicals during normal metabolic processes. However, cells also produce antioxidants that neutralize these free radicals. In general, the body is able to maintain a balance between antioxidants and free radicals.

[0005] [3] An imbalance of free radicals and antioxidants in the body results in a condition called oxidative stress, which can cause damage to cells and tissues and plays a role in the aging process, including in hair and scalp skin.

[0006] [4] Damage to the hair strand, caused by excess free radicals in the hair's proteins, lipids, and melanin, can include cuticle wear, brittleness, roughness, frizz, dryness, dullness (loss of shine), or color fading (melanin damage), and even protein and lipid loss. On the scalp, premature aging occurs, such as damage to hair density, increased porosity, damage to follicle anchoring, and worsening of hair loss and premature graying.

[0007] [5] Several factors can contribute to oxidative stress, especially environmental factors such as pollution, smoke inhalation, and radiation exposure, especially in large urban centers. [6] Uncontrolled oxidative stress in the hair region can accelerate the aging process and contribute to the development of a number of undesirable conditions, such as loss of pigmentation, hair loss, among others.

[0008] [7] Several plant-based active or bioactive ingredients have been studied, as they can also exert antioxidant protective effects in addition to their direct antioxidant effect. Thus, the search for new ingredients or ingredient complexes capable of effectively combating damage caused by oxidative stress and combating free radicals (ROS) caused by UV radiation and pollution in hair and scalp skin continues in the state of the art.

[0009] BRIEF DESCRIPTION OF THE FIGURES

[0010] [8] Figure 1 shows the analytical calibration curve for the HTA method that aims to quantify ROS generated by UV radiation.

[0011] [9] Figure 2 shows the increase in free ROS in different hair types: natural brown, natural blonde, and bleached blonde. The increase in fluorescence indicates an increase in the generation of OH radicals with an increase in the radiation dose.

[0012]

[0010] Figure 3 shows the generation of free OH- radicals (ROS) in the hair strand by UVA radiation, where the effect of antioxidant actives can be observed (natural brown hair).

[0013]

[0011] Figure 4 shows the analytical calibration curve for the lipid peroxidation method.

[0014]

[0012] Figure 5 shows the influence of treatment with antioxidant actives on the oxidation of hair lipids caused by UVA radiation.

[0015] DESCRIPTION OF THE INVENTION

[0016]

[0013] The present invention relates to an antioxidant cosmetic complex, comprising active ingredients from Euterpe oleracea (açaí), Theobroma grandiflorum (cupuaçu), Theobroma cacau (cocoa) and recombinant silk protein.

[0017]

[0014] The antioxidant cosmetic complex according to the present invention prevents damage and combats oxidative stress caused by free radicals, as well as combats and prevents damage caused by sun exposure and pollution to the hair and scalp skin.

[0018]

[0015] The bioactive ingredients according to the present invention can be obtained by processes known in the prior art, from specific parts of plant species. Without imposing any limitation, for example, they can be obtained from aerial parts of plants, such as stems, leaves and flowers, or only leaves, as well as from agro-industrial residues such as pits, mesocarp fiber or mucilage.

[0019]

[0016] According to a preferred embodiment of the present invention, the recombinant silk protein is the recombinant ADF-4 protein which is commercially known by the name Silkgel®.

[0020]

[0017] The active ingredient of Theobroma grandiflorum is an ester, particularly cupuacuate of myristyl instillate or seed of Theobroma grandiflorum.

[0021]

[0018] The ingredient Theobroma cacao is particularly an extract.

[0022]

[0019] In a particular embodiment, the present invention relates to an antioxidant cosmetic complex, to act against damage caused by oxidative stress and combat free radicals (ROS) caused by UV radiation and pollution, in the hair strands and skin of the scalp, in the skin and / or scalp, which comprises:

[0023] (a) 0.001% to 5% Euterpe oleracea, particularly 0.1% to 0.9%;

[0024] (b) 0.001% to 5% of Theobroma grandiflorum, particularly 0.1% to 5%;

[0025] (c) 0.001% to 5% recombinant silk protein, particularly 0.1% to 0.5%;

[0026] (d) 0.001% to 5% of Theobroma cacao, particularly 0.1% to 0.9%.

[0027]

[0020] The percentages indicated above, as well as those indicated in the following examples, are related to the total weight of the final cosmetic composition in which the complex will be conveyed.

[0028]

[0021] In another embodiment, the present invention also comprehensively contemplates cosmetic compositions comprising the antioxidant cosmetic complex according to the present invention together with cosmetically acceptable excipients. The compositions may be of the leave-in or rinse-off type.

[0029]

[0022] The cosmetically acceptable excipients according to the present invention are those known to the person skilled in the art for the composition of cosmetic bases in various forms, for example, emulsions, creams, gels, serums, and others known to the person skilled in the art. For example, without any limitation, the cosmetically acceptable excipients can be selected from those cited in the “International Cosmetic Ingredient Dictionary & Handbook”, 16th Edition.

[0030]

[0023] In another embodiment, the present invention also deals with the use of the antioxidant cosmetic complex for manufacturing a cosmetic composition to act against damage caused by oxidative stress and combat free radicals (ROS) caused by UV radiation and pollution, in hair strands and scalp skin.

[0031]

[0024] Still, in another embodiment, the present invention contemplates a method for topical cosmetic treatment of damage caused by oxidative stress and free radicals (ROS) caused by UV radiation and pollution, which comprises applying to the hair strands and scalp skin, whether or not in need, an effective amount of the complex or cosmetic composition according to the present invention. Thus, the method treated herein may be for cosmetic prevention or treatment.

[0032]

[0025] The following examples, without imposing any limitation, are particular embodiments of the present invention, as well as demonstrating the effectiveness of the complex according to the present invention in treating damage caused by oxidative stress and free radicals.

[0033] EXAMPLES

[0034] Example 1 - Prevention of the damaging effects of free radicals generated by UV radiation on hair using the HTA method

[0035]

[0026] The method consists of the reaction of free radicals (OH-) formed in situ during UV radiation react with terephthalate acid (TA), producing the fluorescent compound: 2-hydroxyterephthalate (HTA).

[0027] The concentration of 2-hydroxyterephthalate (HTA) generated in situ is measured by fluorescence spectroscopy. To calculate the ROS generated in situ, we raised the following calibration curve with different concentrations of terephthalic acid and used the equation of the line as shown in figure 1: y = 1 E + 16x + 103624 / R 2 = 0.9965.

[0036]

[0028] From the calibration curve, we evaluated hair of different colors and degree of damage in the face of increasing doses of UVA radiation (5, 10 and 20 J / cm2), as shown in figure 2 below, and we noticed that the generation of ROS in the strands depends mainly on the dose of UVA radiation to which the strands are exposed and little on the color or degree of damage in the hair.

[0037]

[0029] Straight brown hair irradiated with UVA (20 J / cm2) and treated with different active ingredients in leave-on (without rinsing) and rinse-off (rinsing) applications were evaluated for the ability of the active ingredients to prevent ROS from causing damage to the hair.

[0038]

[0030] It was observed that a 70% reduction in the generation of ROS in situ by the antioxidant action of the active ingredients açaí and cocoa versus placebo and polysilicone 15 (Lubrizol®) as a reference, which is known in the market as sun protection for hair. The result obtained from the combination of active ingredients (acaí + cocoa) in a leave-on and rinse-off application regime stands out, as shown in Figure 3, where the reduction of n.mols of OH generated by UVA is observed: (a) conditioner base with 0.5% cocoa: -62% ROS; (b) conditioner base with 0.5% açaí: -69% ROS and (c) conditioner base cl 0.5% açaí + 0.5% cocoa: -70% ROS.

[0039]

[0031] The data show statistically significant relevance with the differentiation of groups. Statistical evaluation of non-parametric data by the Kruscall-Wallis test with pairwise comparisons by the Dunn procedure (p<0.05).

[0040] Example 2 - Lipid Peroxidation - TBA (thiobarbituric acid)

[0041]

[0032] The prevention of the effects of ROS on hair strands was demonstrated by quantifying the degradation of lipids caused by UVA radiation (20J / cm2), an effect known as lipoperoxidation. For this evaluation, the TBARS method also quantifies (n moles) lipids oxidized by the reaction with thiobarbituric acid (TBA) using fluorescence spectroscopy, thus having the ability to measure total oxidized lipids (lipoperoxidation) in hair strands.

[0042]

[0033] The method consists of measuring lipid peroxidation is determined by the reaction of malondialdehyde (MDA) with thiobarbituric acid (TBA) to form a fluorimetric product MDA+TBA (lex=532 nm / lem=553 nm) as shown in the following reaction:

[0043] MDA TBA MDA-TBA Adduct

[0044]

[0034] The calibration curve shown in figure 4 was constructed with malonaldehyde (MDA) at different concentrations and the calibration equation used to calculate the number of moles of oxidized lipids and evaluate lipoperoxidation in hair.

[0045]

[0035] The evaluation of the active ingredients and mixtures (blends) for the protection or prevention of the effects of ROS generated by UVA radiation (12 J / cm2) and consequently the prevention of lipoperoxidation can be seen in figure 5. Again, the mixture (blend) of active ingredients (acai + cocoa) in leave-on and rinse-off applications protected the hair with 86% reduction in lipoperoxidation in leave-on application with improved effects when compared to the results of the separate active ingredients and the placebo. The results in number of moles of LPO oxidized (reduction) were: (a) base with cocoa: 41%, (b) base with acai: 61% and (c) base with acai + cocoa: 86%.

[0046] Example 3 - degradation of hair proteins due to pollution

[0047]

[0036] Another known source of free radicals (ROS) is environmental pollution, which, through particle deposition or contact in gaseous form with substrates, ends up promoting the degradation of proteins, among other components, in the case of hair strands.

[0037] To evaluate the degradation of hair proteins by pollution, we used the Bicinchoninic Acid (BCA) protein kit assay method from Sigma Aldrich's, which quantifies the degradation of degraded proteins (ug / ml) in hair by UVA / IS spectroscopy, using albumin at different concentrations for the analytical calibration curve. The statistical evaluation of non-parametric data was performed by the Kruscall-Wallis test with paired comparisons by the Dunn procedure (p<0.05).

[0048]

[0038] The results showed that pollution causes considerable protein degradation in the hair strand, increasing protein loss by 3x. The placebo base (without active ingredients) offers little protection to the hair strands. The active ingredients act to protect the hair and, especially when combined, have an enhanced effect for Silkgel + cupuaçu, both in leave-on and rinse-off applications, protecting the hair strand by preventing protein degradation to values ​​lower than the basal protein loss (i.e., hair strand as it is without pollution).

[0049]

[0039] In summary, the results show that ROS (OH.) are generated by the exposure of hair strands to UVA radiation (20J / cm2) and that free radicals promote the oxidation of hair lipids, known as lipoperoxidation, which is known to promote mass loss, damaging the structure of the strands, leaving them stiffer and drier.

[0050]

[0040] Pollution, when deposited on the hair, also generates ROS that degrade the hair proteins, leading to protein mass loss, and the combined active ingredients (Silkgel + cupuaçu) have an effect on protecting the hair from degradation and protein loss.

[0051]

[0041] Thus, the combination of active ingredients (açaí + cocoa + cupuaçu + Silkgel) is powerful in protecting hair strands against free radicals generated by environmental conditions to which hair is exposed daily, such as pollution and UVA radiation.

[0052]

[0042] In addition to the hair strands, the scalp skin is also exposed to these environmental conditions and needs protection because overexposure of the scalp skin generates enough ROS to cause damage to the scalp skin that can lead to hair loss.

[0053] Example 4 - Scalp Skin Assessment

[0054]

[0043] Just as ROS (free radicals) generated by environmental conditions damage hair strands, they also damage the scalp. Known skin effects that extend to the scalp include the loss or reduction of connective tissue, where supporting proteins such as collagen and elastin are located, and dermal cells, especially fibroblasts. Excess ROS due to external or internal factors causes an imbalance in the activity of enzymes called proteases. These proteases include metalloproteinases, such as MMP-1, among others, which act in the degradation of collagen and elastin. Their activity is influenced by the oxidative cascade and inflammatory mediators. Among the important mediators for containing the inflammatory response and tissue damage are IL-10 and transforming growth factor beta (TGF-β).IL-10 acts to balance the inflammatory response by reducing the production of reactive oxygen intermediates, nitric oxide and adhesion proteins.

[0055]

[0044] To evaluate the assets, we chose to focus on two mechanisms: the degradation of skin support proteins, for example, collagen, evaluating MMP-1, and the balance of the inflammatory response of dermal tissues with IL-10, both evaluated against external stimuli such as UVA radiation and environmental pollution.

[0056]

[0045] Normal human dermal fibroblasts (Gibco, Thermo Scientific) and human keratinocytes - HaCaT (Cell Bank of Rio de Janeiro - BCRJ) were seeded in 75 cm bottles 2, grown and expanded in a humidified incubator at 37°C in the presence of 5% CO2, using a specific culture medium. Upon reaching 70-80% confluence, the cells were trypsinized and seeded separately in 6-well plates for subsequent incubation with the test product and evaluation of MMP-1 and IL-10.

[0046] The cells were pre-incubated for 24 hours with three non-cytotoxic concentrations of each product, separately. The test product was used at concentrations of 0.1 mg / mL; 0.05 mg / mL; 0.01 mg / mL. After the pre-incubation period, the cells were subjected to an adverse condition corresponding to their study group, i.e., exposure to cigarette smoke or exposure to 3J of UVA radiation. In parallel, the untreated control of each group was also subjected to stress, and a basal group remained under normal culture conditions. After stress, cells were maintained in a cell incubator for an additional 48 hours.

[0057]

[0047] Metalloproteinase-1 (MMP-1) was quantified using a commercially available sandwich immunoenzymatic assay kit (ELISA) (R&D Systems, USA). Readings were performed in a microplate reader at 450 nm. Metalloproteinase-1 levels were expressed in pg / mL and calculated from reference values ​​obtained with a standard curve constructed with known concentrations of recombinant human MMP-1 standard.

[0058]

[0048] The cytokine IL-10 was quantified using a commercially available enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, USA). The reading was performed in a microplate reader at 450 nm. The cytokine levels were expressed in pg / mL and calculated from the reference values ​​obtained with a standard curve, constructed with known concentrations of the recombinant cytokine.

[0059]

[0049] The evaluation was carried out in monolayer cell culture of keratinocytes and fibroblasts for the active açaí in different concentrations and pureoxin.

[0060]

[0050] From the results for fibroblasts, it was possible to observe that the açaí samples (0.1 mg / mL, 0.05 mg / mL and 0.01 mg / mL) were able to promote a significant reduction (P<0.01) in the synthesis of metalloproteinase-1 (MMP-1) of 16, 12, 11%, respectively, in culture of human fibroblasts subjected to environmental pollution stress, when compared with the environmental pollution control group.

[0051] The tested samples were able to promote a significant reduction (p<0.01) in the synthesis of metalloproteinase-1 of 16, 12, 11, 17 and 12%, respectively, in culture of human fibroblasts subjected to environmental pollution stress, when compared with the environmental pollution control group.

[0061]

[0052] The tested samples were able to promote a significant increase (p<0.01) in the synthesis of metalloproteinase-1 of 16, 14, 19 and 32% in cultures of human keratinocytes subjected to environmental pollution stress when compared with the environmental pollution control group.

[0062]

[0053] The evaluation of the effects of UV radiation on the scalp skin was carried out in monolayer cell culture of human keratinocytes and fibroblasts for the active ingredient açaí in different concentrations and in comparison with the active ingredient Pureoxin benchmark (INCI name: oligopeptide-4), indication for use of 1 to 2%, from Lipotrue, the antioxidant protection action against UV for facial skin.

[0063]

[0054] The person skilled in the art will readily know how to evaluate, through the teachings contained in the text and in the examples presented, the advantages of the invention and propose variations and equivalent alternatives for implementation, without departing from the scope of the invention, as defined in the attached claims.

Claims

CLAIMS 1. ANTIOXIDANT COSMETIC COMPLEX, characterized by comprising active ingredients from Euterpe oleracea (açaí), Theobroma grandiflorum (cupuaçu), Theobroma cacau (cocoa) and recombinant silk protein.

2. COMPLEX, according to claim 1, characterized by comprising: (a) 0.001% to 5% Euterpe oleracea', (b) 0.001% to 5% of Theobroma grandiflorum', (c) 0.001% to 5% recombinant silk protein; (d) 0.001% to 5% Theobroma cocoa.

3. COMPLEX, according to claim 2, characterized by comprising: (a) 0.1% to 0.9% of Euterpe oleracea', (b) 0.1% to 5% of Theobroma grandiflorum', (c) 0.1% to 0.5% recombinant silk protein; (d) 0.1% to 0.9% Theobroma cocoa.

4. COMPLEX according to any one of claims 1 to 3, characterized in that the recombinant silk protein is the recombinant protein ADF-4.

5. COMPLEX, according to any one of claims 1 to 3, characterized in that the active ingredient of Theobroma grandiflorum is an ester.

6. COMPLEX, according to claim 5, characterized in that the active ingredient of Theobroma grandiflorum is myristyl cupuacuate.

7. COSMETIC COMPOSITION, characterized by comprising the antioxidant cosmetic complex, as defined in any one of claims 1 to 6, together with cosmetically acceptable excipients.

8. USE OF THE ANTIOXIDANT COSMETIC COMPLEX, as defined in any one of claims 1 to 6, characterized in that it is in the manufacture of a cosmetic composition to act against damage caused by oxidative stress or combat free radicals (ROS) caused by UV radiation and pollution, in hair strands and / or scalp skin.

9. METHOD FOR COSMETIC TREATMENT, characterized by consisting of applying to the hair strands and / or scalp skin a complex as defined in any one of the claims

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