RO134809A2 - Cosmeceutical composition with photo-protective and regenerative effect - Google Patents

Abstract

The invention relates to a dermatological composition with photo-protective and regenerative effect. According to the invention, the composition consists of 1...5% cationic liposomes associated with fish protein hydrolysate having a content of minimum 25 mg/ml peptides and oily carrot extract with a content of minimum 1.5 mg/ml beta-carotene, 2...10% lanolin and zinc oxide, respectively, 0.3...1% concentrated extract of grapefruit seeds and 75...95% usual natural cream base, the percentage being expressed by mass.

Description

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COSMECEUTIC COMPOSITION WITH PHOTOPROTECTIVE AND REGENERATION EFFECT

Authors: Oana Craciunescu, Daniela Ilie, Alexandra Gaspar-Pintiliescu, Andreea losageanu, Anca Oancea

The present invention relates to a cosmeceutical composition for topical use, with photoprotective and regenerative effect based on nanoformulated fish protein hydrolyzate, biologically active compounds from oily extract of carrot, lanolin and zinc oxide, intended for the protection of healthy skin and reduction of induced skin erythema. of solar radiation, specifically UV type A and B radiation.

It is known that the skin acts as a barrier against external harmful factors of a chemical nature (chemicals, cosmetic ingredients, etc.), of a physical nature (solar radiation, UV radiation, ionizing or gamma radiation, laser light, etc.), mechanical damage, exposure to pathogenic microbial agents, etc. Also, the skin prevents the loss of essential biological fluids, plays an important role in the body's thermoregulation and in the immune system defense. The destruction of this barrier at the level of the epidermis or at most up to the superficial layer of the dermis requires temporary coverage, and the stem, epithelial cells and keratinocytes are able to regenerate the epidermis.

It has been reported that, after prolonged exposure to UV radiation, oxidative stress is generated which is involved in the destruction of skin cells, causing changes in DNA, proteins and lipids (Mohania D. et al., Adv. Exp. Med. Biol ., 996, 71-87, 2017). Reactive oxygen species (ROS) are chemically reactive molecules, derived from molecular oxygen, that are found naturally in its metabolism. Examples of ROS are hydrogen peroxide, superoxide anion and hydroxyl radical. They have an extremely unstable configuration and, therefore, react quickly with other molecules that they turn into free radicals, triggering a chain of reactions that results in DNA damage and lipid peroxidation. The resulting lipid peroxides decompose to form more complex and reactive compounds, such as malondialdehyde and 4-hydroxynonenal.

Currently, there is an alarming case worldwide related to prolonged exposure to the sun or UV radiation, which can induce skin damage through burns (erythema), premature skin aging, wrinkles, photosensitization, decreased immunity or even skin cancers (Pândei R. et al., ISRN Dermatol., 2013, 930164, 2013). Therefore, dermatologists recommend the use of sunscreen measures to prevent harmful effects and the inclusion of compounds with the effect of absorption or reflection of UV radiation in all cosmetics designed to protect skin cells. It is proposed to make cosmetics, as a new type of product, representing a hybrid between a pharmaceutical product for skin conditions and a cosmetic product, which is applied externally and aim at both

improving health as well as beautifying the skin (Martins A. et al., Mar. Drugs, 12, 1066-1101, Ό14).

In the case of defects that reach the deep level of the dermis, a more complex treatment is needed. Thus, aging skin is thinner due to the decrease in collagen content in the dermis. Studies to date have shown that co-treatment with collagen peptides and vitamin C stimulated the synthesis of type I collagen in vivo, reduced the secretion of superoxide oismutase 1 and stimulated the migration and proliferation of fibroblasts (Shibuya S. et al ., Biosci. Biotechnol. Liochem., 78, 1212-1220, 2014). All of these effects have resulted in an increase in skin thickness; some.

Another major problem in skin cells is the balance of ROS secretion. Many Terites have shown that collagen (gelatin) polypeptides from marine sources have the ability to protect olagen and lipids in skin exposed to UV radiation (Chen T. et al., J. Ocean Univ. China, 15, 711718, 2016). One of these antioxidant peptides from fish gelatin, with a molecular weight of -92 Da was identified, sequenced and showed a high capacity to neutralize hydroxyl radicals, with an IC50 inhibitory concentration value of 22.47 pg / ml ( Sun L. et al., J. Funct. Foods, 5, 154-162, 2013). In addition, oral administration of gelatin hydrolyzate helped suppress the act of UV radiation on the skin by inhibiting the activity of endogenous antioxidant enzymes, suppressing NF-kB expression and NF-kB-mediated pro-inflammatory cytokine expression (Chen?. & Hou H. , J. Photochem. Photobiol. B Biol., 162, 633-640, 2016). However, the safety and: clinical instability of oral administration of marine collagen peptides may be impaired due to their inferred origin, which may trigger an immune response by activating the NADPH-oxidase-mediated Toll-like TLR-4 receptor and, implicitly, the overproduction of ROS.

To increase the effectiveness of sunscreen formulas, extracts derived from medicinal plants, rich in compounds with strong antioxidant activity, such as jcoferols, flavonoids, phenolic acids, nitrogen-based compounds and monoterpenes (Korac RR & ihambholjal KM, Pharm. Rev ., 5, 164-173, 2011). Unlike synthetic antioxidants, these extracts have little or no toxicity, thus having a minimal influence on human health and the environment. Natural dyes obtained from fruit and vegetable waste, such as anthocyanins, chlorophylls, carotenoids, lycopene, iboflavin and xanthophylls, also play an important role (Upadhyay RK, Int. J. Green Pharm., 12, S93-S107, 2018). A ¹ -line study showed that the combined administration of marine collagen peptides and antioxidants remarkably improved skin elasticity and sebum production and decreased oxidative destruction 'by Luca C. et al., Oxide. Med. Cell. Longev., 2016, 4389410,2016).

Currently, there are approaches in the literature that propose research and development of liposome formulas to improve the performance of cosmetics. Liposomes • enhance the local release of active ingredients and reduce side effects by reducing the required dose. They can be selected depending on the size of the diameter, the number of bilayers (unilamellar, 'multilamellar), lipid composition and surface charge (neutral, positive or negative) (Bozzuto G.; «Molinari A., Int. J. Nanomedicine, 10 , 975-999, 2015). These carrier-type systems are very useful in the case of parenteral administration of macromolecules with a high content of hydrophobic components, such as peptides and proteins of therapeutic interest (Martins S. et al., Int. J. Nanomedicine, 2,595-607, 2007).

It has been reported to obtain cosmetic / pharmaceutical products based on collagen from fish> i.arin or its hydrolyzate and to improve their mechanical properties by crosslinking with physical agents (US 6,660,280 Bl, 9 Dec. 2003). However, no interaction tests were performed with skin cells, and studies evaluating cosmetic formulas containing fish collagen confirmed the improvement of skin hydration and firmness in a short time for serum products and after repeated applications for those of cream type (Xhauflaire-Uhoda E. et al., Int. J. Cosmet. Sci., 30, 131-138,2008). Sun I. et al. describe cosmetic recipes that contain peptides from marine fish, with or without hyaluronic acid, and report improving the appearance of the skin in mice, especially by reducing the size of the Hours. In human volunteers, a progressive decrease in pore size was demonstrated over 4 weeks of use (CN102178636 (A), 2011-09-14). Another invention describes a lotion for solar irotection that contains, in addition to chemical absorption agents UVA and UVB and oceanic biological substances, such as active peptides and polysaccharides in algae (CN102302417 (A), 2012-01 4). The composition tested on guinea pigs irradiated with a UV lamp for 6 hours showed a role of moisturizing the skin and an activity of reducing inflammation and repairing damaged skin. On human volunteers, a sun protection factor (SPF) was calculated between 22 and 51.

In recent years, new biotechnologies and genetic engineering techniques have been developed for the development of bioactive peptides or the hydrolysis of proteins of interest for therapeutic applications. "Shavandi A. et al., Adv. Food Nutr. Res., 87, 187-254, 2019). However, they have a high non-molecular weight, short half-life, high rate of elimination by degradation into isymatic fluids, low ability to cross the cell membrane and low availability by intestinal administration. the encapsulation of peptides and proteins of therapeutic interest and their transport through the gastrointestinal tract can cause problems due to the high content of hydrophobic components and the tendency to adsorb on glass or plastic surfaces with the help of which their nanipulation is done.

The present invention aims to solve the technical problem of photoprotection and skin regeneration affected by prolonged exposure to UV radiation or aging processes, in a more efficient way, by creating a new natural cosmetic composition, antioxidants and with a dedicated radiation absorption capacity. UV, obtained by combining a fish protein hydrolyzate with regenerating effect with an extract with supercritical carrot fluids with antioxidant and photoprotective effect,> anoformulated, emollient lanolin and zinc oxide as an efficient filter of UV radiation.

The cosmeceutical composition, according to the invention, is a U / A type emulsion, with neutral pH,. consisting of 1 ... 5% cationic liposomes associated with fish protein hydrolyzate having a content of at least 25 mg / ml peptides and oily carrot extract with a content of at least 1.5 mg / ml beta aroten, 2 ... 10 % lanolin, 2 ... 10% zinc oxide and 0.3 ... 1% concentrated extract of irapefruit seeds, included in 75 ... 95% natural cream base, consisting of flower seed oil. Oral, polysorbate 20, cetearyl alcohol, glyceryl stearate, phenoxyethanol, carbomer, potassium sorbate, sodium hydroxide and distilled water, the parts being expressed by weight. Fish protein hydrolyzate can be chosen from a product obtained from collagen from fish skin or a product obtained from others; fish essays (bones, scales, etc.), with a continuous of at least 85% oligopeptides with molecular weight: nedie of 2000 Yes.

The cosmeceutical composition, obtained in accordance with the present invention, has the following * advantages:

• has natural components whose action is efficient in the process of neutralizing the reactive oxygen species formed at the cellular level;

• it is useful for people who are repeatedly and prolonged exposure to solar and UV radiation, but also for those with healthy skin;

• has a good absorption on the skin, is emollient, moisturizing and contains a UV protective filter;

• is biocompatible and has the ability to regenerate dermal tissue;

• the process of obtaining the cosmeceutical composition is simple and feasible.

Protein hydrolyzate from fish has a double role, regenerating and photoprotective, and egret extracts with high carotenoid content have antioxidant and radioprotective action. These nanoformulated compounds in liposomes and included in creams are distributed on the injured area to form a topical, protective film and are gradually released to penetrate the skin.

To date, numerous marine fish sources have been used, but studies on by-products from freshwater fish processing are insufficient (Ngo D.H. et al., Int. J. Biol.

<acromol., 51, 378-383, 2012). The acid treatment of fish waste has a lower extraction efficiency compared to the enzymatic treatment and leads to the destruction of some essential amino acids, such as Trp, Met, Cys, and Asn and Gin are converted into the corresponding acids, Vsp and GIu (Venkatesan J. et al., Marine Drugs, 15,143-161, 2017). Because they have poor functional properties, the resulting protein hydrolysates are used in the production of low-cost fertilizers or substrate for the production of lactic acid (Gao M.T. et al., Bioresource Technol., 97,. '. 414-2420, 2006). The major disadvantage of alkaline hydrolysis is the production of hydrolysates with a low content of amino acids such as Cys, Lys, Arg, Ser, Thr, Ile, as well as the formation of residues such as mtionin (a cysteine dimer) and lysinoalanine, but Trp remains intact ( Pasupuleti V. & Braun S., n: Protein hydrolyzates in biotechnology, Netherlands, Springer, pp. 11-32, 2010; He S. et al., 'Ood Res. Int., 50, 289-297, 2013; Tavane OL, J. Molec. Catalysis B: Enzym., 90,1-11, 2013). This method is frequently used in industry, but not in biotechnology. Preferred is the enzymatic ratification of fish collagen with neutral enzymes because it is hydrolyzed with better and more defined nutritional, functional and bioactive properties, compared to acid / alkaline hydrolysis, but has higher production costs compared to chemical hydrolysis and not ost raised the ladder (Ngo DH et al., Food Chem., 143, 246-255, 2014; Benjakul S. et al., In:

Antioxidants and functional components in aquatic foods, Chichester, UK, John Wiley & Sons Jd., Pp. 237-281,2014). Acidic enzymes (e.g., pepsin) inhibit bacterial growth, but lead to oroduses with low nutritional values because they destroy Trp (Villamil O. et al., Food Chem., 224,160,71,2017). Bioactive peptides isolated by enzymatic hydrolysis from fish collagen are generally oligopeptides with 3-20 amino acid residues. Those from sources such as tuna,. homon and eel contain a significant amount of His dipeptides, such as camosine (β-alanyl-histidine) and anserine (β-alanyl-1-methylhistidine) (Wu HC et al., Food Res. Int., 36, 949- 957, 003).

The bioactive peptides used were extracted from fish bone waste, resulting from: industrial processing of freshwater fish, by enzymatic hydrolysis with papain and conditioning in the form of white-yellow powder with a content of at least 85% molecular weight oligopeptides average of 2000 Yes. They contain sequences similar to those in human skin proteins, have a high solubility, are biocompatible and have biological activity of protecting the hermetic tissue against UV-B irradiation.

* To improve their absorption in the skin, a mixture of lyophilized lipids was combined with ό aqueous solution of bioactive peptides with a content of at least 25 mg / ml peptides by orthexation and sonication, resulting in cationic liposomes of transferosome type, with a diameter under 500 nm. Thus, liposomes are useful as transport systems for these hydrophilic active ingredients in their central aqueous part. Liposomes of small size and with a structure similar to the membrane of skin cells ^ play the role of efficient transporters through the comos layer of the skin for a greater amount of active principles compared to conventional forms (Ahmadi Ashtiani et al., J. Skin & Stern eiis, 3 , e65815, 2016). At the same time, they allow the increase of the surface / volume ratio, the decrease of the quantity of 4 necessary active principles and the improvement of the efficiency of the cost / quality ratio. Nanoformulation of bioactive eptides is an important advantage in easily crossing the comos layer of; delii, their transport in the cell cytoplasm and their action to prevent cell destruction by UV radiation in a short time.

The concentrated natural carrot extract was obtained by extraction with supercritical carbon dioxide and is presented as an oily liquid, orange-reddish in color, with a characteristic odor. The biologically active compounds in the carrot extract are represented by carotenes, lycopene and flavones. -the content rich in beta-carotene or provitamin A provides the extract with antioxidant properties ,. ptyrid and anti-aging, as well as increased protection of the skin against solar radiation, mainly irin the ability to neutralize ROS (Mizgier P. et al., J. Funct. Food, 21, 133-146, 2016). The diphenols present in the carrot extract contribute significantly to cell renewal, decrease skin citation and prolong the bronze due to its antioxidant, anti-inflammatory, itoprotective and antiangiogenic capacity (Smeriglio A. et al., Phytotherapy, 124, 49-57, 2018). However, carrot extract does not have good stability in the presence of air, beta-carotene reacts rapidly with oxygen and an oxidation process takes place. To improve carotene stability, a mixture of lyophilized pides was combined with the oily solution of carrot extract with a continuous content of at least 1.5 ig / ml beta-carpten by vortexing and sonication, resulting in transferosome-type cationic liposomes. si b 500 nm. Thus, liposomes are also useful as transport systems for hypophilic substances in the external liquid layer, t

Lanolin is a natural ingredient, secreted by the sebaceous glands in the skin of sheep and present on woolly irises. After extraction and filtration, lanolin is in the form of a waxy paste, fine, lemisolid, golden-yellow in color and with a characteristic odor, which provides the creams with soft, protective and moisturizing properties. Due to its rich content in ceramides and fatty acids and natural physical characteristics, lanolin is easily absorbed into the skin, reduces water loss through the skin; », at the same time, promotes the penetration into the skin of active ingredients. Anolin-based products are used to treat the skin in cases of atopic dermatitis and various allergies to cosmetics, as well as to restore the skin's natural barrier (Fransen M. et al., Contact) ermatitis, 78, 70-75,2018) .

Zinc oxide is a fine, high-purity, odorless white powder. It is used in cosmetics both as a white pigment and as an antiseptic, purifying and stimulating agent for skin induction. It is an excellent protective filter against UV radiation A and B and improves the coverage of cosmetics (Kim K.B. et al., J. Toxicol. & Environ. Health, 20,. 55-182, 2019). Zinc oxide is very useful in the care of irritated or acne-prone skin, due to its "low osteoporosis, efficacy and lack of side effects (Cervantes J. et al., Dermatol. Ther.,> 1, 2018, two: 10.1111 / dth.l2576 ). It is known for forming a protective barrier against delirium, preventing the multiplication of bacteria on the skin, being antibacterial, antifungal and antiseptic.

The following are two embodiments of the invention:

Example 1.

To obtain 100 g of cosmeceutical product, combine 0.25 mg of lyophilized lipid powder, 0.2; fish protein hydrolyzate, 1 g concentrated carrot extract, 2 g zinc oxide, 4 g lanolin, 0.6 g concentrated grapefruit kernel extract and natural cream base up to 100 g.

The process of obtaining the cosmeceutical composition consists in keeping the lanolin in the water bath at 40 ° C until it melts, then it is melted over the natural base of the cream in the form of an emulsion. U-type gel, consisting of fatty substances and water, previously homogenized with zinc oxide and mixed continuously at room temperature. Separately, the lyophilized powder of lipids containing cholesterol, stearylamine and phosphatidylcholine, in a weight ratio of 1: 2: 7 is mixed with 1 ml of distilled water in which the protein hydrolyzate of the fish was dissolved and, respectively, with the oily carrot extract; vortexing at 2000 rpm, for 30 seconds and then at 500 rpm, for 30 minutes, at room temperature, and sonication at 37 ° C, in 3 repeated series of 3x10 min with a break of 1 min, after which it is left at rest, at 37 ° C, for 60 minutes, for homogenization. The suspension of ationic liposomes associated with bioactive peptides and carrot extract is added over the emulsion, stirring continuously for homogenization, and, finally, the concentrated glycerin extract of grapefruit seeds, as a preservative, is added dropwise. again.

The cream obtained is yellowish in color, has a light consistency, with a characteristic smell and pH Teutru. It is packed in 50 ml boxes and stored at temperatures between 4-25 ° C, for 6 units. The cosmeceutical composition has a moisturizing, photoprotective, regenerating and emollient effect, can be applied on healthy or damaged skin due to prolonged exposure to UV radiation and prevents skin aging by anti-radical action.

Example 2.

To obtain 100 g of cosmeceutical product, combine 0.25 mg of lyophilized lipid powder, 0.1; fish protein hydrolyzate, 0.6 g concentrated carrot extract, 3 g zinc oxide, 6 g lanolin, 0.6 g concentrated i-xtract of grapefruit seeds and natural cream base up to 100 g.

The process for obtaining it is as described in Example 1.

The main ingredients with photoprotective and regenerating role, namely, peptides from fish protein hydrolyzate and biologically active compounds from carrot extract, were tested in terms of antiradical activity, as well as the ability to absorb UV radiation A, B and C. The nano-formulated ingredients were tested for biocompatibility with skin cells, wound healing capacity on an experimental model made by scratching a monolayer of fibroblast cells from the stabilized line NCTC clone L929 and cell radiation protection is a model of fibroblasts exposed to UV-B irradiation. The obtained results showed that both the protein lidrolysate and the carrot extract show antioxidant activity, by neutralizing reactive oxygen species, such as hydroxyl and superoxide radicals, but also free radicals of ABTS and DPPH. Determination of the UV radiation protection factor following the tracing of the spectra of; bsorption in the case of carrot extract, but also of the obtained cream showed a projection action. idicata, both in the UV-B and UV-A domain. Tests on fibroblast cultures cultured in the presence of nanoformulated ingredients indicated a lack of cytotoxicity and stimulation of Ibroblast proliferation and migration, which significantly accelerated the recovery of the cell monolayer in the model. quarries after only 24 hours. In addition, cells treated with nanoformulations and then irradiated showed higher viability compared to untreated ones. Thus, it has been shown that bioactive peptides can penetrate faster through the cell membrane into the cytoplasm to intervene in cellular metabolism, and bioactive plant compounds can eliminate excess ROS due to cell exposure to UV-B ventilation.

Claims (1)

A cosmeceutical composition with photoprotective and regenerating effect, based on nanoformulated bioactive plant peptides and plant compounds, characterized in that it consists of 1 ... 5% cationic hyposomes associated with fish protein hydrolyzate having a continuum of at least 25 mg / ml peptides and oily carrot extract with a continuous of at least 1.5 mg / ml beta-carotene, 2 ... 10% janolin, 2 ... 10% zinc oxide and 0.3 ... 1% concentrated extract of grapefruit seeds, included in ' ⁷ 5 ... 95% natural cream base, consisting of sunflower seed oil, polysorbate 20,; Cetearyl icool, glyceryl stearate, phenoxyethanol, carbomer, potassium sorbate, sodium hydroxide and distilled water, the parts being expressed by weight.