BR102016018124A2 - LOADED NANOPHESES RED PROPOLIS EXTRACT, NANOPHESES PROCESSING, DERMOCOSMETIC COMPOSITIONS CONTAINING THE SAME AND USES - Google Patents

Abstract

The present invention provides nanobeads loaded with red propolis extract (npv), composed of standardized hydroalcoholic extract of red propolis from fresh red propolis, and nanocapsulant, dispersing, stabilizing polymeric matrices, preventing self-oxidation of creams. Suspension or solid state npv are considered intermediate products for the production of dermocosmetic compositions. The invention also deals with the characterization processes of nanospheres loaded with nanometer sized red propolis extract, as well as the identification of isoflavonoids and antioxidant activity and leishmanicidal action. Additionally, the present embodiment provides processes for obtaining the npv-containing dermocosmetic compositions, preferably in the form of nonionic creams, and may also be in the form of cationic creams. Finally, uses of dermocosmetic compositions containing nanopheres loaded with red propolis extract in the treatment of leishmaniasis, healing and antioxidant are presented.

Description

(54) Title: NANO SPHERES LOADED RED PROPOLIS EXTRACT, PROCESS OF OBTAINING NANO SPHERES, DERMOCOSMETIC COMPOSITIONS CONTAINING THE SAME AND USES (51) Int. Cl .: A61K 35/644; A61K 8/98; A61P 17/18; A61P 33/02; A61P 29/00; (...) (73) Holder (s): FEDERAL UNIVERSITY OF ALAGOAS, APÍCULA FERNÃO VELHO (72) Inventor (s): TICIANO GOMES DO NASCIMENTO; MÁRIO CALHEIROS DE LIMA; LAIS FARIAS AZEVEDO; PRISCILLA DA FONSECA SILVA; IRINALDO DINIZ BASÍLIO JÚNIOR; MARIA CRISTINA DELGADO DA SILVA; JOSEALDO TONHOLO; CAMILA BRAGA DORNELAS; EDUARDO JORGE DA SILVA FONSECA; CÍRIA VIEIRA BARBOSA; ANA RÚBIA BATISTA RIBEIRO; CANTÍDIO FRANCISCO DE LIMA FILHO; ISABEL CRISTINA CELERINO DE MORAES PORTO; MARIANNA PORFÍRIO BRANDÃO; LOUISIANNY GUERRA DA ROCHA (57) Abstract: The present invention features nanospheres loaded with red propolis extract (NPV), composed of standardized hydroaloolic extract of red propolis from fresh red propolis, and polymeric matrices with a nanoencapsulating, dispersing, stabilizing function. , avoid self-oxidation of creams. Suspended or solid NPVs are considered intermediate products for the production of dermocosmetic compositions. The invention also deals with the characterization processes of nanospheres loaded with nano-sized red propolis extract, as well as the identification of isoflavonoids and antioxidant activity and leishmanicidal action. Additionally, the present modality of invention presents processes for obtaining dermocosmetic compositions containing NPVs, preferably in the form of non-ionic creams, and may also be in the form of cationic creams. Finally, uses of dermocosmetic compositions containing nanospheres loaded with red propolis extract are presented in the treatment of leishmaniasis, healing and antioxidant.

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Descriptive report of the invention patent for “Nanospheres LOADED WITH RED PROPOLIS EXTRACT, PROCESS OF OBTAINING Nanospheres, DERMOCOSMETIC COMPOSITIONS CONTAINING THE SAME AND USES”.

[001] The present invention belongs to the field of natural products useful in public health in aesthetics and beauty, more specifically those of nanoparticle systems loaded with active principles of red propolis extract with biological activities, intended for administration in the dermal and mucous membranes. Nanospheres loaded with red propolis extract (NPV), processes for obtaining NPV, and dermocosmetic compositions containing NPV and uses are proposed. The production of NPV is linked to the use of a standardized hydroalcoholic extract of red propolis, an active principle of NPV, and a system of matrices of organic pharmaceutical polymers with an encapsulating, dispersing and stabilizing function of these suspended nanospheres, which will be nanoprecipitated and lyophilized to obtain nanospheres. polymeric compounds containing red propolis extract in solid state that will be used for the production of dermocosmetic compositions. Finally, the dermocosmetic compositions containing NPEPV have application in dermocosmetic and cosmeceutical sectors, in aesthetics, beauty treatment, in reparative surgeries, due to their antioxidant, anti-inflammatory, anticancer and healing activities, as well as for their leishmanicidal action.

BACKGROUND OF THE INVENTION [002] The constituents of propolis have been identified worldwide (Pietta, P .; Gardana, C .;

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Scaglianti, M .; Simonetti, P., Journal of Pharmaceutical and Biomedical Analysis, V. 45, p. 390-399, 2007). Flavonoids, phenolic acids and terpenes are the main substances found and used to track quality and, in some cases, to demonstrate the authenticity of propolis from some geographic regions (Volpi, N .; Bergonzini, G., Journal of Pharmaceutical and Biomedical Analysis, V. 42, pp. 354-361, 2006). Among the flavonoids and phenolic acids most commonly used as markers, the following can be mentioned: quercetin, canferol, naringenin, chrysin, pinocembrine, galangin (flavonoids); gallic acid, caffeic acid, p-coumaric acid, ferulic acid, cinnamic acid and derivatives (phenolic acid), clerodanes (diterpenoids) (Rosalen, PL; Castro, ML; Cury, JA, Quimica Nova, V. 30, N. 07 , 1512-1517, 2007); (Yao, L .; Jiang, Y .; Singanusong, R., Datta, N .; Raymont, K., Food Chemistry, V. 86, p.169-177, 2004); (Weston, R. J., Mitchell, K. R., Allen, K. L., Food Research International, V. 37, p. 166-174, 2004).

[003] The increasingly popular use of propolis and its derivatives with antimicrobial action (Sforcin, JM; Fernandes Jr., A .; Lopes, CAM; Bankova, V. And Funari, SRC, J. Ethnopharmacology, V. 73, p. 243-249, 1998), anti-inflammatory (Khayyal, MT; el-Ghazaly, MA; el-Khatib, AS, Drug Experimental and Clinical Research, V. 19, p. 197-203, 1993), antiviral (Vynograd, N .; Vynograd, I .; Sosnowisky, Z., Phytomedicine, V. 7, p. 1-6, 2000), anticarcinogenic (Bazo, AP; Rodrigues, MAM; Sforcin, JM; Camargo, JLV, Ribeiro, JL; Salvadori, DMF, Teratogenis, Carcinogenis and Mutagenis, V. 22, p. 183-194, 2002) and immunomodulatory (Sforcin, JM, Kanemo, R. Funari,

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S. R. C., Journal of Venemous Animal and Toxins, V. 8, p. 19-29, 2002); (Sá-Nunes, A .; Faccioli, L.H., Sforcin, J. M., Journal of Ethnopharmacology, V.83, p. 93-97, 2003) has been demonstrating the great therapeutic power of propolis to replace conventional synthetic drugs.

[004] The characterization of all these biological activities associated with the tendency to use natural products has resulted in an increased demand for propolis and products containing propolis, such as extracts, creams, ointments, tablets, capsules, or powders (H. Menezes, M Bacci Jr., S.

D. Oliveira, F. C. Pagnocca, Antibacterial properties of propolis and products containingpropolis from Brazil, Apidologie, V. 28, p. 71-76, 1997).

[005] In the case of red propolis, this was classified as the 13th subtype of Brazilian propolis found in the mangroves, lagoons, rivers and beaches of northeastern Brazil between the states of Sergipe, Alagoas, Pernambuco and Paraíba by the researcher (Andreas Daugsch, Cleber S. Moraes, Patricia Fort and Yong K. Park, Brazilian Red Propolis — Chemical Composition and Botanical Origin, eCAM 2008; 5 (4) 435-441 doi: 10.1093 / ecam / nem057). The main botanical origin of red propolis is due to the Dalbergia ecastophyllum plant, locally known as Rabo-deBugio, which has a red resinous exudate released from its sap.

[006] National and international researchers with expertise in propolis have been identifying the constituents of red propolis. 11 isoflavonoids and 1 chalcone of Cuban red propolis were identified and among these substances we can mention: Formononetina (Biochanina B), Bichanina A, vestitol, ortho-methyl vestitol, medicarpine,

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4/57 homopterocarpine, liquiritigenin and [007] region other pterocarpan derivatives, isoliquiritigenin (chalcone) (Anna Piccinelli et al. J. Agric. Food Chem. 2005, 53, 9010-9016). Other subclasses of flavonoids have also been identified, including new types of flavonoids from the red propolis of the Northeast region, which can be mentioned: rutin, quercetin, luteolin, pinocembrine and biochanin A, formononetine, daidzein, liquiritigenin, pinobanksina, dalbergine (neoflavoneides) (Andreas) , Cleber S. Moraes, Patricia Fort and Yong K. Park, Brazilian Red Propolis— Chemical Composition and Botanical Origin, eCAM, Evidencebased complementary and alternative medicine, 2008; 5 (4) 435441 doi: 10.1093 / ecam / nem057).

Researchers studying the red propolis from the mangroves of Marechal Deodoro-Alagoas, identified the flavonoid chrysin and quercetin, new isoflavonoids in the red propolis of Alagoas, the vestitol, 2'4'dhydroxymethyl flavana, isoprenylated benzophenones, as well as phenolic acids and phenolic esters and phenolic compounds. , methyleneugenol, guiacol, dimethyl esters of butanenedioic acid, hexadecanoic acid methyl esters (Bruno B. Silva, Pedro L. Rosalen, Jaime A. Cury, Masaharu Ikegaki, Vinícius C. Souza, Alessandro Esteves and Severino M. Alencar Chemical Composition and Botanical Origin of Red Propolis, a New Type of Brazilian Propolis, eCAM, evidence based complementary and alternative medicine 2008; 5 (3) 313316, doi: 10.1093 / ecam / nem059).

[008] Additionally, researchers identified 14 substances in the red propolis of the mangroves of Alagoas, among them triterpenes (cycloartenol, lupeol, elemicin, αamirine and β-amirine), phenolic acids (anethole, eugenol and

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5/57 methylphenol), isoflavonoids (isosativan, medicarpine) (Boryana Trusheva, Milena Popova, Vassya Bankova, Svetlana Simova, Maria Cristina Marcucci, Patrícia Laguna Miorin, Flavia da Rocha Pasin and Iva Tsvetkova, Bioactive Constituents of Brazilian Red Propolis, eCAM, evidence-based complementary and alternative medicine 2006; 3 (2) 249-254, doi: 10.1093 / ecam / nel006).

[009] With a view to discovering molecules and their therapeutic targets for curing diseases, red propolis has been subjected to several detailed studies on its biological activities. Research on the identification and structural elucidation of new substances continues to be carried out in the identification of new substances present in the red propolis extract. A new chalcone, 3, 4, 2, 3'tetrahydroxchalcona and a new C-glycosylated flavonoid, narigenin-8-C-hexoside, were identified, in addition to (3S) -vestitol and (3S) -7O-methylvestitol (Adne A Righi, Thiago R Alves, Giuseppina Negri, Lucas M Marques, Henrique Breyer and Antonio Salatino, Brazilian red propolis: unreported substances, antioxidant and antimicrobial activities, J Sei Food Agríc 2011; 91: 2363-2370, DOI 10.1002 / jsfa.4468) . Biological research with the ethanol extract of red propolis and isolated substances, vestitol and neovestitol, demonstrated anti-inflammatory activity in the concentration of 10mg / kg and antimicrobial activity in concentrations between 6 to ΙΟΟμρ / πΛ (Bruno Bueno-Silva, Severino M. Alencar, Hyun Koo, Masaharu Ikegaki, Gil VJ Silva, Marcelo H. Napimoga, and Pedro L. Rosalen, Anti-inflammatory and Antimicrobial Evaluation of Neovestitol and Vestitol Isolated from Brazilian Red

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Propolis, J. Agric. Food Chem. 2013, V. 61, p. 4546-4550, dx.doi.org/10.1021/jf305468f).

[010] Red propolis extracts enriched with xanthochimol and formononetine have been showing in vitro antitumor activities against different cell lines such as melanoma using cell line B16F10, ATCC CRL6475 (Estela Maria Novak, Martha Silveira and Costa Silva, Maria Cristina Marcucci, Alexandra Christine Helena Frankland Sawaya, Begona Giménez-Cassina López, Maria Angela Henriques Zanella Fortes, Ricardo Rodrigues Giorgi, Kamila Tamie Marumo, Rosangela Felipe Rodrigues, Durvanei Augusto Maria. Antitumoural activity of Brazilian red propolis fraction enriched with xanthochymol and formononetin: An in vitro and in vivo study, Journal of Functional Foods, 11, (2014), 91102).

[011] Ethanol extract of red propolis has been showing activity against cancer cells of the glioblastoma (SF-295), ovary (Ovcar-8) and colon (HCT-116) lines (Izabel Cristina Gomes de Mendonça, Isabel Cristina Celerino de Moraes Porto, Ticiano Gomes do Nascimento, Naiana Soares de Souza, José Marcos dos Santos Oliveira, Rodolfo Elleson dos Santos Arruda, Kristiana Cerqueira Mousinho, Aldenir Feitosa dos Santos, Irinaldo Diniz Basílio-Júnior, Abhishek Parolia, Francisco Stefânio Barreto, Brazilian red propolis : phytochemical screening, antioxidant activity and effect against cancer cells, BMC Complementary and Alternative Medicine (2015) 15: 357.DOI 10.1186 / s12906-015-0888-9).

[012] Proteomic identification studies of laryngeal cancer cell markers have also demonstrated activity of red propolis extract against cell lines

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7/57 epidermoid laryngeal against ATCC Hep-2 strains (Caroline Olivieri da Silva Frozza, Tanara da Silva Ribeiro, Gabriela Gambato, Caroline Menti, Sidnei Moura, Paulo Marcos Pinto, Charley Christian Staats, Francine Ferreira Padilha, Karine Rech Begnini, Priscila Marques Moura de Leon, Sibele Borsuk, Lucielli Savegnago, Odir Dellagostin, Tiago Collares, Fabiana Kommling Seixas, João Antonio Pêgas Henriques, Mariana Roesch-Ely, Proteomic analysis identifies differentially expressed proteins after red propolis treatment in Hep-2 cells, Food and Chemical Toxicology 63 (2014) 195-204).

[013] Research around the world exists to cure negligible diseases such as leishmaniasis and extracts of propolis has also been showing activity against these parasites. Duran et al. (2011) (Duran, N., Muz, M., Culha, G., Duran, G., Ozer, B. 2011. GC-MS analysis and antileishmanial activities of two Turkish propolis types. Parasitology research, 108, 95- 105,) showed leishmanicidal activity for 2 types of propolis extracts from Turkey (Hatay propolis and Bursa propolis) and demonstrated an IC50 of 250pg / mL and 500pg / mL, respectively. Another relevant study by Duran et al. (2008) (Duran, G., Duran, N., Culha, G., Ozcan, B., Oztas, H., Ozer, B. 2008. In vitro antileishmanial activity of Adana propolis samples on Leishmania tropica: a preliminary study Parasitology Research, 102, 1217-1225) showed good results for Adana Propolis against leishmania at concentrations of 250pg / mL. Another type of Turkish propolis extract (Kayseri propolis) studied by Ozbilge et al. (2010) (Ozbilge, H., Kaya, E.G., Albayrak, S., Silici, S. 2010. Anti-leishmanial activities of ethanolic extract of Kayseri propolis. African Journal of

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Microbiology Research. 4, 556-560.) Demonstrated leishmanicidal activity against Leishmania tropica with an IC50 of 32pg / mL. Comparative study of Brazilian green propolis extracts and Bulgarian propolis extracts carried out by Machado et al. (2007) showed activity against leishmanicides 4 different species of Leishmania chagasi (amazonensis braziliensis and Major). Brazilian green propolis extract showed an IC50 close to 49pg / mL against braziliensis chagasi and Major species, while the Bulgarian propolis extract showed leishmanicidal activity for amazonensis, chagasi and Major species with an IC50 between 2.8 pg / mL and 41.3pg / ml. Brazilian red propolis extract was evaluated in macrophages infected with Leishmania amazonensis, and demonstrated that the concentration of 25 ug / mL is able to increase the reducing activity of MTT being active against intracellular parasites present in macrophages (Ayres, DC, Marcucci, MC, Giorgio, S. 2007. Effects of Brazilian propolis on Leishmania amazonensis. 39 Memórias Instituto Oswaldo Cruz, 102, 215-220.).

[014] The presence of various flavonoids and phenolic acids in propolis extracts and tinctures shows that these substances act synergistically in cytostatic / cytotoxic, anti-inflammatory, healing, antimicrobial and antioxidant action. In this way, the cocktail of combined substances, even in low concentrations, will promote a potent action against pathogens. The development and production of herbal and herbal medicines has been regulated in the country with the resolutions that establish quality control methodologies for these products. Standardization studies of phytotherapy and opotherapies recommend tests in order to guarantee the quality of these

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collection, phytochemical screening, qualitative and quantitative determination of phytochemical markers, authenticity testing, processing, production, stability study, validation of analytical methodologies, packaging and product quality control.

[015] Due to its different characteristics in terms of chemical composition and multiple biological activities, as well as for presenting characteristics of production and standardized management in the mangrove areas of Alagoas-Brazil, respecting a policy of environmental preservation and conservation of the areas of Atlantic forest and mangroves, the production of fresh red propolis and hydroalcoholic extract is part of a productive chain of opotherapeutic products that promotes the sustainable development of the mangrove and lagoon region and is awarded the geographical indication seal with the designation of origin.

[016] Development of new drug delivery systems or active substances on certain therapeutic targets is a necessity to achieve therapeutic success or to demonstrate physiological health benefits in preventing diseases or curing them or even with aesthetic and beauty action , and it is as important as the discovery of molecules with a specific therapeutic action. Thus, in the present invention different dermocosmetic compositions were planned in the semi-solid form for topical use or in the mucous membranes, preferably cutaneous topical and modified release.

[017] In the development of new systems for the release of active extracts containing various active substances with

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10/57 polimultifunctional activity in dermocosmetics it is necessary to have the knowledge of the skin as an organ with a barrier function against chemical and physical agents, the permeation pathways and penetration factors in the skin, as well as the main cosmetic excipients responsible for the permeation of these substances active in the skin and attached organs. The skin functions as a large organ that covers a surface greater than 20,000 cm ² with varied properties and functions, as in addition to protecting the body from the invasion of microorganisms, it protects against UV rays, produces melanin and immunoglobulins, in addition to controlling body temperature, blood pressure, as well as having sensory function. The anatomical knowledge of the skin is necessary when choosing the active extract encapsulation systems, as well as cosmetic excipients that will compose a dermocosmetic composition. The skin has several histological layers, being divided into stratum corneum, epidermis, dermis and subcutaneous adipose tissue. The stratum corneum and hair follicles and glands (sweat and sebaceous) are considered the first barriers that limit the speed of entry of the active substances that make up a cosmetic cream (Leon Lachman, Herbert A. Lieberman, Joseph L Kanig, Theory and Practice in Industry Pharmaceuticals, Vol. II, chapter 18, pages 907-953, 2001).

[018] In dermocosmetics, the choice of encapsulating agents of the active extract and excipients used in cosmetology with multifunctional properties is extremely important, that is, excipients with various functions such as: skin permeation facilitators, photoprotective action, specific cosmetic action, skin stabilizers lipophilic / hydrophilic system, viscosity control and

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11/57 sensory, in addition to being biocompatible and biodegradable. Cosmetic excipients are essential in a dermocosmetic composition, as they have characteristics that ensure stability to the composition, facilitate administration, promote the release of the active substances in the dermal matrix, thus promoting the bioavailability of the active substances in the different layers of the skin and consequently the appropriate pharmacological action without toxicity, as well as consumer acceptability from a sensory and aesthetic point of view. For the development of semi-solid systems with application in cosmetology it is necessary to have a set of excipients that will properly release the drug or active substances from a dermocosmetic matrix and among them are the excipients: emulgents, emollients, humectants, water absorbent bases, hydrophilic bases , waxes, hydrocarbons, oilseeds, alcohols, fatty acids, esters, polyols, insoluble powder, essences, preservatives, antiseptics. Some excipients used in cosmetic compositions can alter the internal structure of the stratum corneum by changing the barrier function and increasing the rate of permeation of active substances by the stratum corneum, among which we can mention: water, propylene glycol, oleic acid, urea, DMSO, laurocapranos, pyrrolidones, and other surfactants (Leon Lachman, Herbert A. Lieberman, Joseph L Kanig, Theory and Practice in the Pharmaceutical Industry, Vol. II, chapter 18, pages 907-953, 2001).

[019] From the point of view of the process and the costs of production, pharmacotechnical incompatibilities, whether chemical or physical, between active substances and cosmetic excipients must be avoided. Some

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12/57 cosmetic excipients can generate negative influences on the dermocosmetic composition. Cosmetic compositions due to the presence of lipid excipients, or the presence of lipid active substances, are subject to auto-oxidation reactions with further decomposition, being an inconvenience for these formulations, resulting in unpleasant aroma and appearance and rancification reactions. Therefore, the choice of excipients must be well defined to avoid problems of drug-excipient or excipient-excipient incompatibility, reducing the stability of the active substance (s), in the composition, as well as toxicity problems. Some active substances can be highly reactive in a dermocosmetic composition and produce the most diverse chemical reactions, reducing the stability of these semi-solid compositions. Among the chemical reactions commonly predicted are the hydrolysis reactions of esters, amides, imides, lactone; oxidation reactions due to the presence of a high concentration of oxygen in the composition and auto-oxidation; isomerization reactions; photodegradation reactions initiated by the action of light causing a decrease in the potency of the active substances (Katsunori Yoshida, Tomoko Sekine, Fumiaki Matsuzaki, Toshio Yanaki *, and Michihiro Yamaguchi. Stability of Vitamin A in Oil-InWater-In-Oil-Type Multiple Emulsions JAOCS, Vol. 76, No. 2 (1999) PP.6); degradation reactions with the formation of toxic substances such as formaldehyde in the cosmetic composition and generating possible toxic effects such as atopic dermatitis in consumers of these products (Margareta Bergh, Kerstin Magnusson, J. Lars G. Nilsson and Ann-Therese Karlberg. Formation of formaldehyde and peroxides by air oxidation of

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13/57 high purity polyoxyethylene surfactants. Contact Dermatitis,

1998, 39, 14-20).

[020] In general, propolis extracts present in their composition several active substances of different phytochemical classes, namely: flavonoids, phenolic acids, terpenes, clerodans, xanthones, benzophenones, among others, that can in contact with different dermocosmetic excipients to promote chemical reactions of more diverse, reducing power, stability, in addition to resulting in unpleasant looking products and still resulting in toxic effects. In this sense, it is necessary to develop techniques for micro or nanoencapsulation of active substances of natural origin.

[021] Micro and nanoencapsulation techniques are used by the pharmaceutical, food and cosmetic industries in order to solve technological bottlenecks, including: protecting the active substance (s) against extrinsic agents (moisture, light, oxygen) that can cause reactions of hydrolysis, photodegradation, oxidation; prevent the loss of volatile substances; increase the shelf life of the product; promote controlled release of active substances; as well as improving specific delivery characteristics of active substances in target tissues.

[022] Nanoparticles, in general, are obtained by various preparation techniques, among which we can mention the techniques of nanoemulsion formation, and among them we can mention: double emulsification method, emulsification-coacervation method, polymer coating method , layer-by-layer coating method, emulsification-diffusion method and nanoprecipitation method (Mora14 / 59

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Huertas, C. E. Fessi, H. Elaissari, A. Polymer-based nanocapsules for drug delivery. International Journal of

Pharmaceutics 385 (2010) 113-142).

[023] However, these methods of obtaining suspended nanoparticles, if not properly prepared, can present disadvantages such as the risk of microbiological contamination due to being in an aqueous medium, degradation of the polymer by hydrolysis, physicochemical instability due to the aggregation of the particles or decrease biological activity of the incorporated active substances. Some techniques for preparing nanoparticles prevent microbial contamination, polymer degradation problems and possible instabilities; so the use of nanoparticle drying processes is one of the resources to guarantee substance stability for long periods.

[024] There are nanoparticle drying techniques, such as evaporator evaporation techniques, spray-dryer drying, others. A widely used method for microencapsulated and nanoencapsulated is the freeze drying method. Lyophilization is widely used in the drying of thermolabile substances, such as material containing protein substances, microorganisms such as lactic bacteria used in the production of yeasts, edible fungi, among others. It is also used as a method of encapsulating active materials sensitive to temperature and high pressure. The drying of nanoparticles has the advantage of increasing the shelf life of active substances, avoiding degradation of substances in aqueous and suspended media, facilitating the redispersion and reincorporation process in semi-solid systems.

solvent by freeze-drying and obtaining

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15/57 [025] To date, there are some state-of-the-art documents with potential for innovation for propolis microencapsulation and few documents for propolis nanoencapsulation, but so far no state of the art documents with red propolis nanoencapsulation have been identified. for dermocosmetic purposes. The following documents will cite them.

[026] Some documents in the state of the art demonstrate the use of lyophilization to obtain microencapsules. KR1020030063053 produces granules from a mixture of 10 to 30% propolis, 40 to 70% lactose, 0.1 to 1.0% vitamin and 10 to 30% dehydrated honey by freeze-drying. Chinese document CN1711992 obtains propolis microcapsules using a high-cost excipient, beta-cyclodextrin, followed by a lyophilization process. JP2006028174, on the other hand, produces granules of propolis using a mixture of 50% amino acids (propolis extract: mixture of amino acids). Microencapsules of propolis containing a modified starch (Octenyl-starch succinate) and gum arabic were obtained by the Spray-Dryer technique (Silva FC, Thomazini M., Alencar SM and Favaro-Trindade CS, XVIIth International Conference on Bioencapsulation, Groningen, Netherlands ; September 24-26, 2009). The obtaining of propolis microencapsules for various purposes is described in the prior art. For example, CN102920652 prepares microencapsules of propolis with other biopolymers: chitosan, hydroxypropyl beta-cyclodextrin and sodium glycerophosphate for the treatment of periodontal pathologies. CN102772596 uses multi-blend of plant extract, propolis and 5%

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16/57 chitin to obtain preparations using the spraydryer technique with application in the treatment of obesity. CN102641256 obtains propolis microencapsules with urea, gelatin, paraffin, and toxic solvent (hexane) for preparing microencapsules with a large particle size between 70 to 150pm. CN102429141 uses a gelatin / pectin and propolis system to prepare slow-release microcapsules using lyophilization.

[027] Process of obtaining propolis components in the dry state using gelatin containing propolis solution from the city of Maringá (Paraná) were prepared by spray-drying. The optimization of spray drying conditions and the proportions of gelatin and mannitol were investigated (M.L. Bruschi, M.L.C. Cardoso, M.B. Lucchesi, M.P.D. Gremião, International Journal of Pharmaceutics (2003) 264, 45-55). It is essential to highlight that the use of different types of excipients allows different mechanisms for the release of the active substances in the pharmaceutical matrix (microencapsulated), in addition to different physiological actions. The Brazilian patent application PI05001757 uses only two excipients (gelatin and mannitol) and demonstrates only the process of preparing an intermediate matrix containing extracts of propolis, without disclosing the origin of the propolis used. Patent applications WO 2014/186851 and BR 10 2012 013590-6 made by our research and innovation group with opotherapeutic drugs, in particular with red propolis, make use of gelatin, pregelatinized starch and colloidal silicon dioxide for the preparation of microencapsulated Brazilian red propolis with gastroresistant release. French patent application FR 725623 is aimed at a system

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17/57 multi-microparticulate galenic prolonged release of different active ingredients. These microcapsules are formed by an active ingredient core, coated with a covering based on, for example, ethyl cellulose, polyvinylpyrrolidone, Castor oil and magnesium stearate. European patent EP 548356 claims a multiparticulate fast-breaking tablet comprising an active substance in the form of microcrystals or microgranules provided with a cover. WO 2014/166994 A1 develops a mixed active ingredient release system and comprises a particulate nanomicro system that presents active ingredients inside the nanoparticle, as well as inside the microparticle with application in compositions for oral use. [028] Few nanotechnological products and processes for preparing nanoparticles using propolis have been developed worldwide, by different research groups, in different ways and for different purposes (nutraceutical, therapeutical as well as cosmeceutical). A polymeric nanoparticle formulation of propolis (nano-food propolis) was developed using micellar aggregates of cross-linked and random copolymers of N-isopropylacrylamide (NIPAAm) with N-vinyl-2-pyrrolidone (VP) and poly (ethylene glycol) monoacrylate (PEG) (Kim, DongMyung, Lee, Gee-Dong, Aum, Seung-Hyun, Kim, Ho-Jun, Biological & Pharmaceutical Bulletin, (2008) 31, 1704-1710). American patent US 20130295181 and patent EP2633862 of the same patent family develops propolis nanocapsules based on polyvinyl alcohol (PVA), submicrometic particle size (<1000nm) between 220nm to 438nm with different applications and

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18/57 different areas: agronomic, pharmaceutical and cosmetic, etc. Patent family WO / 2011/001181 and CA2765920 uses propolis extract to obtain a solid carrier matrix of active substances, specifically develops a solid film containing propolis.

[029] Some examples show the specific applicability in the field of cosmetics using coated nanoparticles containing hydrophilic and / or oleophilic substances in dermocosmetic compositions, as well as in oral compositions. WO2015 / 022471 A1 uses coating nanocapsules for active oleophilic substances present in UVA / UVB sunscreens. KR20100078349 discloses preparations of propolis nano emulsions using excipient β-cyclodextrin. Patent document WO 2015/003155 A1 shows the applicability of biocompatible nanofibers containing chitosan, honey, acetic acid, gelatin, collagen, alginates, BSA and other forming agents of reticulated nanometric networks and natural or synthetic active substances against various pathologies hypertension, healing in

Patent document PCT / US2013 / 075714 deals with compositions containing anhydrous powder containing liquid submicrometric particulate system (<1000nm) used for the preparation of emulsions, suspensions, solutions and powders for cosmetic or pharmaceutical purposes. This system consists of a hydrophilic core containing a hydrophobic coating that will protect all active ingredients in its core. Chinese patent document CN103520069 uses honey and propolis in compositions and beauty products for nails including compositions like gel, creams and adhesives. Soft gelatin capsules were developed in the patent document as bacteria, wounds, cancer.

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19/57 to maintain beauty and youth, as well as regulate immunity, cholesterol and senescence. Chinese patent document CN1031160399 develops soap with natural extracts, propolis and base ingredients for the manufacture of soap for the multifunctional treatment of cleaning, removing bacteria, reducing allergies and antiseptic. Chinese patent CN101461846 develops a gel containing plant extract and propolis against herpes zoster. Japanese patent document JP2008247830 uses propolis extract and amaranth extract in cosmetic composition to inhibit skin blemishes, pigmentations, age spots. CN101181198 patent document uses kojic acid, fibroin, propolis and other ingredients in the preparation of a nourishing beauty cream for removing skin blemishes. Invention patent uses honey, pearl powder, propolis, glycerin to produce a multifunctional massage and beauty cream reducing inflammatory skin processes such as acne. JP2006016373 invention patent document with multifunctional property of weight reduction, hair reduction and beauty treatment uses propolis, royal jelly, plant extracts, and collagen. KR1020030093890 uses propolis extract and green tea extract and other plant extracts to produce a soap with antibacterial, antiseptic, polymultifunctional action, atopic dermatitis and odorizing action. Japanese patent JP2003055138 uses honey and Brazilian rosemary propolis extract to produce a type of plaster with antibacterial and antifungal action, as well as for beauty treatment. Chinese invention patent CN1244386 uses propolis and perilla extract to produce particles between

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0.01 to 10gm, in compositions used in the treatment and prevention of caries, periodontal inflammation, antibacterial agent.

[030] In the process of nanoencapsulation of drugs and active substances, some polymers with nanoencapsulating properties have been used in compositions called nanospheres containing a solid core or nanocapsules containing an oily or liquid core. A combination of nanoencapsulating polymers has also been used to form so-called solid polymeric matrices (nanospheres) or liquid core nanocapsules coated by a solid wall. The term nanoencapsulated can also be used more broadly without defining the type of polymeric nanoparticle (nanosphere or nanocapsule). The use of polymeric systems or also called polymeric matrices has been found in the scientific literature, being able to perform the combination of one or more of the following polymers: Span, polyvinyl alcohol, polyethylene glycol, polyoxypropylene, Tween, pluronic, polyoxyethylene, polycaprolactone, ceramide, acid polylactic / polyglycolic, alpha-tocopherol polyethylene glycol succinate, trimethyl chitosan chloride, among others (M. Wulff-Perez, A. Torcello-Gomez, MJ Galvez-Ruiz, A. MartinRodriguez, Stability of emulsions for parenteral feeding: Preparation and characterization of o / w nanoemulsions with natural oils and Pluronic F68 as surfactant, Food Hydrocolloids 23 (2009) 1096-1102 .; Hearan Suh, Byeongmoon

Jeong, Ramesh Rathi, Sung Wan Kim, Regulation of smooth muscle cell proliferation using paclitaxel-loaded poly (ethylene oxide) -poly (lactide / glycolide) nanospheres, J Biomed Mater Res. 1998 Nov; 42 (2): 331-8. ; Régis Coco,

Laurence Plapied, Vincent Pourcelle, Christine Jérôme, David

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J. Brayden, Yves-Jacques Schneider, Véronique Préat, Drug delivery to inflamed colon by nanoparticles: Comparison of different strategies, International Journal of Pharmaceutics 440 (2013) 3-12).

[031] Use of polymeric coating matrices guarantees advantages to the formulator, among them, stability to the nanoparticulate matrix system, prevents problems of degradation or reactions of hydrolysis, photodegradation, among others, and also guarantees a standardized process for the release of active substances. So far some research shows the use of polymeric matrices in the process of micro or nanoencapsulation of drugs with pharmacological activity against cancer (tamoxifen, paclitaxel), using plant extract, propolis extract, as well as phytopharmaceuticals (rutin) (Juliana S. Almeida , Fernanda Lima, Simoni Da Ros, Luis OS Bulhões, Leandro M. de Carvalho, Ruy CR Beck, Nanostructured Systems Containing Rutin: In Vitro Antioxidant Activity and Photostability Studies, Nanoscale Res Lett (2010) 5: 1603-1610 .; Jugminder S Chawla, Mansoor M. Amiji, Biodegradable poly (o-caprolactone) nanoparticles for tumor targeted delivery of tamoxifen, International Journal of Pharmaceutics 249 (2002) 127-138; Ezequiel Bernabeu, Gustavo Helguera, Maria J. Legaspi, Lorena Gonzalez, Christian Hocht, Carlos Taira, Diego A. Chiappetta, Paclitaxel-loaded PCL-TPGS nanoparticles: In vitro and in vivo performance compared with Abraxane®, Colloids and Surfaces B: Biointerfaces 113 (2014) 43- 50 .; Alexandru Luca, Betul Cilek, Vasif Hasirci, Serpil Sahin, Gulum Sumnu, Storage and Baking Stability of Encapsulated Sour Cherry Phenolic Compounds Prepared from Micro- and NanoSuspensions, Food Bioprocess Technol (2014) 7: 204-211).

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22/57 [032] So far, a polymeric system has been detected containing polymeric matrix polycaprolactone and polyvinyl alcohol (PCL-PVA) and a particular type of propolis from southeastern Brazil (Mogi das Cruzes-SP), with a preparation process by emulsification technique, followed by evaporation. However, the particles obtained were presented as microparticles with particle size between 5 to 10pm (N. Duran, P. D. Marcato, C. M. S. Buffo, M. M. M. De Azevedo,

E. Esposito, Poly (e-caprolactone) / propolis extract: microencapsulation and antibacterial activity evaluation, Pharmazie 62 (2007) 4, 287-290). Patent family US20130295181 and EP2633862 develops propolis nanocapsules based on polyvinyl alcohol (PVA) with sizes ranging from 220nm to 438nm with different applications including agronomic, pharmaceutical and cosmetic areas.

[033] None of the described patent documents reveals the obtaining of polymeric nanoparticles containing red propolis extract (NPEPV) using the PCLpluronic polymeric system (F-108) through a nanoprecipitation method followed by centrifugation and freeze drying to obtain solid polymeric nanoparticles with application in the preparation of dermocosmetic creams. Thus, the modalities of the invention described in this document have considerable advantages in view of the state of the art.

[034] The invention proposed in this document seeks to fill a technological gap in terms of the application of red propolis as an active element in modified-release dermocosmetic compositions for the treatment and prevention of diseases, proposing a form of modified release of nanoparticles polymeric containing

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23/57 red propolis extract (NPEPV). In this way, polymeric nanoparticles containing modified propolis red propolis extract, processes for obtaining NPEPV, dermocosmetic compositions with NPEPV, processes for preparing dermocosmetic compositions with NPEPV and uses of these dermocosmetic compositions with antioxidant, anti-inflammatory, cytotoxic / cytotoxic action are proposed. , wound healing in the postoperative period, decubitus wounds, aesthetic and beauty treatments, hygiene and cleaning.

[035] It should be noted that the use of red propolis is a determining characteristic in innovation, in terms of new health technologies with the use of propolis and opoterapics, since, red propolis has a characteristic composition, which will have a primary function in the activity of NPEPV and other dermocosmetic compositions, for the prevention and treatment of diseases, aesthetics and beauty. [036] The development of an analytical method for the identification and quantification of isoflavonoids in the extract and in the polymeric nanoparticles containing red propolis extract by high performance liquid chromatography coupled to the diode array detector also appears as an innovative element of this document.

[037] Still, as an essential differential in view of the state of the art, polymeric nanoparticles containing red propolis and dermocosmetic compositions proposed in the present invention, use encapsulating, stabilizing and dispersing compounds in the intermediate matrix that will promote controlled release, as well as avoid oxidative processes between phenolic compounds present in red propolis with other cosmetic excipients present in the external matrix, in addition to

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24/57 a poly-multifunctional oil / aqueous external matrix with accessory activities that contain biocompatible, biodegradable, non-toxic and stable cosmetic excipients with rheological characteristics for modified-release cosmetic / cosmeceutical semisolids. In this way, the composition disclosed here can be used to solve semi-industrial and industrial technical problems in the dermocosmetic areas, as well as some applications for therapeutic use.

SUMMARY OF THE INVENTION [038] The present invention features nanospheres loaded with red propolis extract (NPV), composed of standardized red propolis extract from the raw material red propolis, and three nanopolymer matrix systems composed of coating and dispersant / stabilizer polymers . These nanoencapsulating systems consist of a binary or ternary system, with biocompatible, biodegradable properties and promoting a modified release of the active principle, protecting the dermocosmetic compositions from oxidative attack of the phenolic substances present in the active extract of red propolis, in addition to presenting leishmanicidal activity. The invention also deals with the process of obtaining suspensions of polymeric nanoparticles that are nanoencapsulated in a modified release system, by means of the nanoprecipitation technique followed by centrifugation and lyophilization to produce solid nanospheres loaded with red propolis extract. In addition, this modality of invention is presented in terms of semi-solid compositions containing NPV, preferably in the form of

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25/57 non-ionic cream, non-ionic gel-cream, non-ionic gel-cream, non-ionic lotions, and can also prepare cationic creams. The disclosed composition comprises: i) a core containing red propolis active substance (s) combined with a polymeric nano-coating matrix that can modify or delay the release of the active substance (s) contained in the red propolis extract ; (ii) an intermediate dispersing / stabilizing layer of the core; (iii) an external oil / aqueous layer responsible for the dilution, dispersion and stabilization of the intermediate layer and core in a dermocosmetic composition, as well as for promoting the dissolution and permeation of the active components through the skin barriers. In addition, the invention deals with the process of preparing the dermocosmetic composition, from the preparation of suspensions of polymeric nanoparticles containing red propolis, as well as from polymeric nanoparticles in the solid state containing red propolis extract. Finally, uses of NPV and dermocosmetic compositions containing NPV with antioxidant and leishmanicidal action are presented.

BRIEF DESCRIPTION OF THE FIGURES [039] The modality of the invention, together with additional advantages of the same, can be better explained and understood by reference to the attached figures and the following description:

[040] The attached Figure 1 shows Electronic Scanning Photomicroscopy of NPV - NCEU20% (A and B), NCEU 12% (C and D) and NCPCL30% (E and F) using colloidal silicon dioxide cryoprotectants (A, B and E) and sodium starch glycolate (C, D and F). Photomicrographs with 3000 times magnification

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26/57 (5pm scale) and magnification up to 12000 times (1pm scale) showing particles in nanometer scale.

[041] The attached Figure 2 shows DSC thermograms of the NPV. NEU compositions (A), NPCL compositions (B) and NEUPCL compositions (C).

[042] The attached Figure 3 shows FTIR-ATR spectra of NPV. Red propolis extract, placebo composition and NEU 30% composition (A and B). Red propolis extract, placebo composition and 30% NPCL composition (C and D). Red propolis extract, placebo composition and NEUPCL composition 30% (E and F).

[043] The attached Figure 4 shows the chromatograms (UPLC-DAD) of the red propolis extract (A), Placebo NEU (B), composition NEU30% (C), composition NPCL 30% (D) and composition NEUPCLC 30% ( E) that identifies flavonoids / isoflavonoids present in the red propolis extract and compositions and absence of flavonoids in the placebo. [044] The attached Figure 5 presents a normalized IC50 determination graph in an in vitro leishmanicidal assay against Leishmania (V.) braziliensis (A). Schematic illustration of some hypothetical biochemical mechanisms of cellular inhibition of Leishmania (V.) braziliensis by the flavonoids present in the red propolis extract of Alagoas as well as in the nanospheres loaded with red propolis extract (B).

[045] The attached Figure 6 shows photos of nonionic creams containing NPCL30% loaded with red propolis extract to obtain a final concentration in the cosmetic cream of 2.0% (A), 1.5% (B), 1.0% ( C) and 0.5% (D) of the red propolis extract. The solid-state NPV nanospheres were

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27/57 directly incorporated into the non-ionic cream or using suspensions of polymeric nanoparticles (B).

DETAILED DESCRIPTION OF THE INVENTION [046] The processes and compositions described in the present invention can be better detailed and understood by reference to the figures present in this document and the following description:

- Nanospheres loaded with Red Propolis Extract (NPV) [047] The nanoparticles (solid or suspended) presented in this document have different concentrations of flavonoids, isoflavonoids, chalcones and phenolic acids due to the specificity of red propolis. Additionally, the nanoparticles in question have an important characteristic of protecting the semisolid matrix system (cosmetic cream) from the pro-oxidizing action of flavonoids and phenolic compounds present in the red propolis extract, since cosmetic creams prepared directly with the red propolis extract undergo darkening (brown color) in up to 10 days, due to oxidizing reactions of phenolic compounds against the cosmetic excipients present in this dermal release matrix system. It is also worth mentioning that the proposed polymeric coating has a second important functionality, of modified release of the active compounds, delaying the release of flavonoids / isoflavonoids, for a longer duration of action, in addition to maintaining the active principles for longer with biological activity in the layers. of the skin, being

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28/57 important in treatments against skin inflammation, cancer, leishmanicidal action, wound healing in leishmaniasis, in addition to promoting anti-aging action due to its antioxidant properties.

[048] The invention modalities of nanospheres loaded with red propolis extract presented in this invention can also be applied for the production of red propolis capsules.

[049] The NPVs, in addition to the extract with the active principles of interest, obtained from red propolis, present pharmaceutical excipients with different functionalities in their composition, namely: active compound (red propolis extract), polymeric coating matrix with nanoencapsulant function , emulsifier, dispersant, stabilizer and antioxidant.

[050] One embodiment of the present invention, a NEU polymeric nanoparticle in different percentages of red propolis extract, presents itself as a composition of: standardized extract of red propolis from the raw material propolis red in a proportion between 5 to 80%, preferably between 10 and 55%; encapsulating agent in a proportion between 50 to 70% and dispersing / stabilizing agent in a proportion between 15 and 30%.

[051] Another modality of the proposed invention, a polymeric NPCL nanoparticle in different percentages of red propolis extract, is presented with the following composition: standardized hydroalcoholic extract of red propolis from the raw material propolis red in a proportion between 10 to 80% , preferably between 15 and 55%; encapsulating agent in a proportion between 40 to 90% and a dispersing / stabilizing agent in a proportion between 1 and 20%.

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29/57 [052] Another modality of the proposed invention, a NEUPCL polymeric nanoparticle in different percentages of red propolis extract, is presented with the following composition: standardized hydroalcoholic extract of red propolis from the raw material red propolis in a ratio of 10 95%, preferably between 15 and 50%; encapsulating agents in a proportion between 30 to 80% and a dispersing / stabilizing agent in a proportion between 1 to 20%.

[053] In the modalities of the invention composed of NEU, all NEU20%, NEU25%, NEU30%, NEU40% and NEU50%, the polymeric matrix system is characterized by being a binary system that promotes nanoencapsulation of the nanoparticle, in addition to stabilization and dispersion after nanoprecipitation in aqueous medium, composed of Eudragit® E100, also known as poly (butyl-co (2-dimethylaminoethyl) methacrylate) (methyl methacrylate) (PM: 47,000) and pluronic F-108 triblock copolymer (PM: In the modalities of invention in NEU20%, NEU25%, NEU30%, NEU40% and NEU50%, the dispersant / stabilizer system can also be characterized by being composed of Eudragit®E100 (PM: 47,000) and pluronic F-68 copolymer triblock (PM: 14.000). [054] Polymeric nanospheres loaded with red propolis extract (NEU20%, NEU25%, NEU30%, NEU40% and NEU50%), as they consist of biocompatible and biodegradable excipients, in addition to providing modified release to the nanoencapsulated, it also reduces the attack oxidative of the phenolic compounds present in the red propolis extract to lipid excipients, which make up the composition of semi-solid forms such as: cationic creams and cream ointment for dermocosmetic purposes, as well as in solid and injectable pharmaceutical forms.

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30/57 [055] In the modalities of the invention composed of NEUPCL, all NEUPCL20%, NEUPCL25%, NEUPCL30% and NEUPCL40%, the polymeric matrix system is characterized by being a ternary system that promotes nanoparticle nanoencapsulation, in addition to stabilization and dispersion after nanoprecipitation in aqueous medium, composed of poly-εcaprolactone or PCL, Eudragit® E100, also known as the poly- (butyl-co methacrylate (2-dimethylaminoethyl) (methyl methacrylate) (PM: 47.000) and pluronic F- 108 triblock copolymer (PM: 14000) In this type of invention in NEUPCL20%, NEUPCL25%, NEUPCL30% and NEUPCL40%, the dispersant / stabilizer system can also be characterized by being composed of poly-s-caprolactone (PM: 10,000), Eudragit®E100 (PM: 47,000) and pluronic F68 triblock copolymer (PM: 14,000).

[056] Still, it is important to highlight that, the polymeric nanospheres of red propolis containing PCL and Eudragit®E100 proposed in this document are innovative because they present manometric particle sizes in an inventive modality and are also presented here and in relation to to other microencapsulated propolis systems, previously mentioned, in addition to being used to solve the problem of cationic creams and cream ointments containing red propolis extract, as well as in solid and injectable dosage forms.

[057] In the modalities of the invention composed of NPCL, all NPCL20%, NPCL25%, NPCL30%, NPCL40% and NPCL50%, the polymeric matrix system is characterized by being a binary system that promotes nanoencapsulation of the nanoparticle, in addition to stabilization and dispersion after

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31/57 nanoprecipitation in aqueous medium, composed of poly-εcaprolactone or PCL, (P.M .: 10,000) and pluronic F-108 triblock copolymer (P.M .: 14,000). In this modality of invention in NPCL20%, NPCL25%, NPCL30%, NPCL40% and NPCL50%, the dispersant / stabilizer system can also be characterized by being composed of poly-caprolactone (PM: 10,000) and pluronic F-68 triblock copolymer (PM: 14,000). The compositions presented here are used to solve the problem of non-ionic creams loaded with red propolis extract, as well as being able to be used in solid and injectable dosage forms.

- Process of obtaining NPV [058] The preparations of polymeric nanoparticles containing red propolis extract are obtained through the superconcentrated and standardized hydroalcoholic extract of red propolis with residual solvent percentage between 0.01 to 10%, preferably between 0.01 to 3% using the maceration process and ethyl alcohol between 75 to 85 ° GL for a period of time between 48 to 72 hours of extraction and concentration of the extract in a rotary evaporator using a temperature of 45 ° C and a pressure of 650 mmHg.

[059] A dark red solid mass, with a yield between 50 to 70%, will be obtained with a percentage of solvent between 0.01 to 35%, being called red propolis extract. The extract should preferably have a percentage of solvent between 0.01 to 3%.

- Preparation of the nanospheres system loaded with red propolis extract (NPV)

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32/57 [060] The steps for obtaining the NPV include: 1) Weighing the components of the organic phase including the standardized extract of red propolis, poly-s-caprolactone (PM 10,000), Eudragit®E100 (PM: 47.000) ) and organic solvent with sufficient volume for solubilization; 2) Weighing of the components of the aqueous phase including the pluronic F-108 triblock copolymer (P.M. 14,000) and ultrapure water of the milli-Q type sufficient to solubilize the pluronic F-108, in a separate container; 3) The components of the organic phase are solubilized in acetone, or chloroform or dichloromethane, preferably in acetone and transferred to an ultrasonic bath with specific power and frequency for complete solubilization for a period of 15 minutes; 4) The components of the aqueous phase are also transferred to an ultrasonic bath under the same conditions as the organic phase for complete solubilization; 5) The components of the aqueous phase are diluted to a volume between 14 to 99 times with milliQ water; 6) The components of the organic phase are transferred to a falcon tube and dispersed with the aqueous phase with a dilution factor between 15 to 100 times, for example, (1 part of the volume of the organic phase: and 99 parts of the volume of the aqueous phase, v: v) to obtain a suspension of polymeric nanoparticles containing red propolis extract; 7) the concentration of solid mass of the red propolis extract, of the nano-coated polymer and of the pluronic F-108 with dispersant function comprises an amount of solid mass between 0.0110 to 0.706% that are dispersed in the ultrapure water type milli- Q (amount of solid mass of the components of the organic and aqueous phases dispersed in milli-Q water); 8) At this moment, the phenomenon of

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33/57 nanoprecipitation with a suspension of polymeric nanoparticles containing red propolis extract; 9) The process continues with the centrifugation of the suspension of polymeric nanoparticles containing red propolis extract using centrifugation speed between 4000 to 6000rpm for a period of 15 minutes and removal of the supernatant; 10) followed by the milliQ water washing step (optional) of the nanoprecipitated polymeric nanoparticles; 11) followed by a new centrifugation process, with disposal of the supernatant; 12) After centrifugation and supernatant disposal, the polymeric nanoparticles nanoprecipitated by the centrifugation process are collected and concentrated in volume between 1 to 10 ml, preferably between 3 to 6 ml to start the physical-chemical characterization process and the freeze-drying process.

- Physico-chemical characterization of polymeric nanoparticle suspensions containing red propolis extract (NPEPV) [061] Nanoparticle suspensions loaded with red propolis extract (NEU20%, NEU30%, NEU40% and NEU50%), in addition to the compositions (NPCL20 %, NPCL25%, NPCL30%, NPCL40%, NPCL50%), as well as the compositions (NEUPCL20%, NEUPCL25%, NEUPCL30%, NEUPCL40%) were characterized using pH techniques, average particle diameter distribution, polydispersion index and zeta potential. To determine the pH, suspensions of polymeric nanoparticles were placed directly in contact with the pH electrode. To determine the other physical-chemical parameters mentioned here, the solution was diluted 4 times with Milli-Q water and analyzed using Malvern's Zetasizer, Nano ZS equipment. The

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34/57 results were expressed as the average of 2 cycles of 20 scans (Table 1).

- Drying of polymeric nanoparticle suspensions containing red propolis extract [062] Nanoparticle suspensions loaded with red propolis extract (NEU20%, NEU30%, NEU40% and NEU50%), in addition to the compositions (NPCL20%, NPCL25%, NPCL30 %, NPCL40%, NPCL50%), as well as the compositions (NEUPCL20%, NEUPCL25%, NEUPCL30%, NEUPCL40%) were subjected to freeze drying on Terroni® equipment, model LD1500 consisting of: 1) vacuum drying chamber containing 3 shelves, 2) a condenser for lowering the temperature to -43 ± 5 ° C and, 3) low vacuum pump. The system pressure was controlled to maintain 300pHg as an indication of complete drying of solid polymeric nanoparticles containing propolis extract.

[063] It is important to note that, before the freeze drying process, the polymeric nanoparticle suspensions were subjected to two main freezing processes which were: A) Slow Freezing, in which the polymeric nanoparticle suspensions were placed in a -20 freezer ° C for a period between 48 to 120 hours and immediately transferred to a freeze dryer to perform the drying process for a period between 24 and 48 hours. B) Quick freezing with the aid of liquid nitrogen at 196 ° C for a period of 10 to 20 minutes. In this process the suspensions of polymeric nanoparticles were placed in antibiotic type bottles with a capacity of 50 mL, which were packed in a stainless steel type box.

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35/57 before receiving the freezing liquid. It is also worth mentioning the use of cryoprotective agents in this particular freezing process, namely: colloidal silicon dioxide between 0.1 to 10%, as well as sodium starch glycolate solution between 0.1 to 30%. In these conditions of sample preparation prior to the drying process, the drying period was reduced between 33 to 50%.

- Physico-chemical characterization of solid polymeric nanoparticles containing red propolis extract [064] The solid nanospheres loaded with red propolis extract (NPV), after drying by freeze drying, were characterized by different analytical techniques to demonstrate that the extract of red propolis was completely coated by the proposed polymeric matrix, and that the nanospheres are in nanometric size even after drying by freeze drying and that the phenolic compounds present in the red propolis extract were encapsulated in percentages above 30%. Among the analytical techniques, we mention: A) Infrared spectroscopy (FTIR-ATR). The matrix system of solid nanospheres loaded with red propolis extract was submitted to FTIR-ATR analysis in the scanning range between 4000 to 400 cm-1 with a number of scans between 64 to 128. The equipment used was Thermo Scientific with software Ommic acquisition. Samples of solid polymeric nanoparticles without propolis extract were also analyzed. B) Thermal Analysis (DSC). The solid nanospheres loaded with red propolis extract (NPV) were subjected to calorimetric analysis in a Shimadzu calorimeter, model DSC-60 under

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36/57 nitrogen with a flow of 50mL / min., At a heating rate of 10 ° C / min. and in the temperature range between 30-400 ° C. A quantity of 2.0mg ± 10% was used and placed in an aluminum crucible which was hermetically sealed. The equipment was calibrated with standards of indium and zinc. Samples of placebo nanospheres (without propolis extract) were also analyzed. C) Analysis of NPEPV Particles Morphology and Mean Diameter using Scanning Electron Microscopy (SEM). The solid nanospheres loaded with red propolis extract (NPV) were subjected to 2 drying methods: Drying by slow freezing, followed by lyophilization technique (method A); quick freeze drying, followed by freeze drying technique (method B), the latter performed with the use of cryoprotective agents. The NPVs in solid state, after drying, were prepared in stubs with double carbon tape or directly in the coverslips, submitted to the metallization system with gold film under 10 mA current for 7 minutes in a Sanyu Electron metallizer, model Quick Coater SC -701 and then analyzed using a Shimadzu Scanning Electron Microscope, model SSX-550 Superscan. D) Identification, Dosage and (%) Degree of Encapsulation of flavonoids present in the red propolis extract and in the NPV. The determination (identification and determination) of the flavonoids present in the red propolis extract and in the NPV was carried out in a UPLC-DAD equipment from Shimadzu, which consists of the following modules: a high pressure pump (Model, LC-20ADXR), degasser (model , DGU-20A3R), autoinjector (model, SIL-20AXR), chromatographic column oven, photodiode array detectors (model, SPDM-20A) and fluorescence detector (model RF37 / 59

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20A), a controller (model, CBM-20A) and Labsolution software from shimadzu. The separation of the flavonoids occurred in a stationary phase (C18, 150 x 4.6 mm; 5pm), a mobile phase that consisted of solvent A (ultrapure water) and solvent B (acetonitrile), pumped in a flow of 0.3 mL / min. in a gradient elution condition, with 70% water and 30% acetonitrile in the beginning with a 40% change in the percentage of organic to 100% acetonitrile in 40 minutes, followed by an isocratic condition with 100% acetonitrile for up to 53 minutes, returning to the condition in 54 minutes, followed by an isocratic condition with 30% acetonitrile for up to 60 minutes. [065] Analytical standards acquired from sigma-aldrich (formononetine, isoliquiritigenin, daidzein, Biochanine A, pinobanksine) and Extrasyntesis (Liquiritigenina) were exactly weighed 2.0 mg and transferred to 10 mL volumetric flasks to obtain a concentration of 200pg / mL. Working solutions were prepared by dilution technique to obtain concentrations of 7.50; 5.00; 2.50; 1.00; 0.50 and 0.15pg / mL and obtain a calibration curve, which was used to determine the content (dosage) of flavonoids in the red propolis extract and in the NPV. The identification of the flavonoids was carried out using retention times comparative to the analytical standards using the same analytical conditions as the working day. The degree of encapsulation study was also carried out under these same analytical conditions and with the 5 markers (Liquiritigenina, pinobanksina, formononetina, isoliquiritigenina, Biochanina A) present in the red propolis extract, which were determined in the NPV and in the red propolis extract following these separation conditions. The red propolis extract (100 mg) in the solid state (<3% solvent) was

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38/57 solubilized in absolute ethanol with the aid of an ultrasonic bath (5 minutes) and transferred to a 10 mL volumetric flask to obtain a concentration of 10 mg / mL. A new dilution step was performed to obtain a 1mg / mL working solution and to make dilutions to 400 and 250qg / mL. These solutions were filtered in 0.45pm filter units and injected (2qL) in the UPLC-DAD system. The NPV containing red propolis extract or NPV without red propolis extract (Placebo Composition) were weighed exactly and corresponding to 1mg / mL of the theoretical concentration of red propolis extract, solubilized with a solvent system (acetone: ethanol, 6: 4, v: v), then the dilution to 310qg / mL was performed with the same solvent system, in order to verify the maximum amount of flavonoid encapsulated in the NPV. The NPVs containing red propolis extract or without propolis extract (placebo) were also weighed exactly and corresponding to 1mg / mL of the theoretical concentration of red propolis extract, solubilized with a solvent system (water: ethanol, 7: 3, v : v), then dilutions to 310qg / mL were performed with the same solvent system (water: ethanol, 7: 3, v: v), in order to verify the amount of flavonoid not encapsulated in the NPV, but present in the outer portion of polymeric nanoparticles (NPV). The degree of encapsulation was determined using the following formula: Degree of Encapsulation (%) = (marker concentration in NPEPV / marker concentration in EBPV) * 100.

- Characterization of the Antioxidant Activity of NPV and Red Propolis Extract using DPPH

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39/57 [066] Solid nanospheres loaded with red propolis extract, solid placebo nanospheres (without red propolis extract) and red propolis extract were evaluated for their antioxidant activities. Stock solutions of 1 mg / mL of the NPV compositions were prepared containing red propolis extract, which were exactly weighed to obtain the corresponding 1 mg / mL of red propolis extract in these compositions and solubilized in an acetone: ethanol solvent system (6: 4 , v: v). The same procedure was performed with red propolis extract. Aliquots of 400pL, 50pL, 25pL and 13pL of the NPV stock solution were transferred to 5 mL volumetric flasks and 2 mL of a 3mM DPPH ethanolic solution added, being left to stand for 30 minutes in the dark for the reaction to occur, and then, the volumetric flask was checked with absolute ethanol to obtain concentrations of 80.0pg / mL, 10.0pg / mL, 5.0pg / mL and 2.5pg / mL. The same procedure was performed with the red propolis extract and with the placebo nanospheres (without red propolis extract).

[067] The samples of red propolis extract were subjected to UV-vis spectrophotometer reading in photometric mode adjusted to a wavelength of 520 nm at the end of the 30-minute period. The NPV and placebo nanospheres showed turbidity at the end of the reaction period, being subjected to a centrifugation with a rotation speed of 5000 rpm for a period of 5 minutes. The supernatant was read on the UV-vis spectrophotometer under the same conditions. Readings were also performed on the 3 mM DPPH reference solution in ethanol, absolute ethanol solution to zero the equipment and the white solutions (ethanol

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40/57 absolute + 3 mM DPPH without the addition of red propolis extract or NPV) to calculate% of antioxidant activity. The calculation of% of antioxidant activity was given by the following formula: (%) Antioxidant activity = 100- (((The sample - A blank) x10 0) / A control). Where: The sample = Absorbances of the NPV samples containing DPPH solution, A blank = Absorbances of the NPV samples without the addition of the DPPH solution, A control = Absorbance of the DPPH reference solution in ethanol.

- Process for preparing dermocosmetic compositions containing NPV [068] The steps for obtaining dermocosmetic compositions containing NPV can be identified by the following main Processes: 1) Process A: Hot preparation - Obtaining a suspension of nanoparticles loaded with red propolis extract ( NEU30% or NPCL30% or NEUPCL30%) in aqueous phase; Heating of the aqueous phase containing the suspension of polymeric nanoparticles containing propolis extract and the oil phase containing self-emulsifying cosmetic excipients of a non-ionic cream in separate containers at a temperature between 75 to 85 ° C, preferably the aqueous phase at 80 ° C and phase oily at 80 ° C; Incorporate the aqueous phase over the oil at the same temperature under agitation between 200 to 400rpm, preferably between 270 to 300rpm, maintaining temperature and agitation constant for 10 minutes; Cool with moderate agitation to 40 ° C; Add a complementary phase (other dermatological actives, sensory modifiers and preservatives) under agitation and allow to cool to room temperature; Check pH and make corrections if necessary for 5.5 to

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6.5. 2) Process B: Cold preparation - Obtaining solid nanospheres loaded with red propolis extract obtained by freeze drying using slow freezing or fast freezing; Obtaining nonionic base creams, gel-cream, non-ionic gel-cream, non-ionic lotions when prepared with NPCL30% and obtaining cationic creams or cream-ointment when obtained with NEU30% and NEUPCL30%; cooling and conditioning of non-ionic cream bases at room temperature; Incorporation of solid polymeric nanospheres (NPCL30%) loaded with red propolis extract in “non-ionic cold base creams by means of geometric dilution technique under mechanical or manual agitation and without the use of organic or lipid solvents for the solubilization of solid nanospheres loaded with red propolis extract in these base creams. Or, Incorporation of solid polymeric nanospheres (NEU30% or NEUPCL30%) loaded with red propolis extract in “cold cationic base creams by means of geometric dilution technique under mechanical or manual agitation and without the use of organic or lipid solvents for solubilization of the solid nanospheres loaded with red propolis extract in these “cationic creams or“ cream-ointment.

[069] The processes A and B described were compared with the preparation in cationic creams containing suspensions of polymeric nanoparticles, as well as with the preparation of “cationic creams containing red propolis extract soluble in organic solvent and without the technological process of nanoencapsulation, which showed physical and chemical incompatibilities after 10 days of preparation with browning of the composition to brown. However,

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42/57 two processes for obtaining dermocosmetic creams containing NPV, the compositions remained compatible and stable for a period longer than 180 days.

[070] The obtained NPV can be used in several dermocosmetic compositions, preferably as semi-solid formulations, such as: non-ionic creams, gel, gel-cream, lotions, non-ionic lotions, as well as powders, makeup powders, skin whitening , facial masks, among others.

- Dermocosmetic Compositions containing NPV [071] The obtained NPV can be used in several dermocosmetic compositions, preferably as semi-solid formulations, such as: non-ionic cream, gel-cream, gel-cream, lotions, non-ionic lotions, makeup powders, skin lightening, facial masks, among others. NPV can also be used in cationic cream formulations and also cream ointment.

[072] The pharmaceutical compositions proposed in this document consist of compositions preferably in the form of non-ionic cream, non-ionic gel-cream, non-ionic gel-cream, non-ionic lotions and also cationic creams, which feature: i) a core containing active red propolis substance (s) combined with polymeric nano-coating matrix that can modify or delay the release of active substance (s) contained in the red propolis extract, as well as protect other excipients cosmetics from the oxidative attack of phenolic compounds present in the propolis extract; ii) a dispersing / stabilizing intermediate layer of the core; iii) an external oil / water layer responsible for

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43/57 processes of dilution, dispersion, and stabilization of the intermediate layer and core in a dermocosmetic composition, as well as for promoting the dissolution and permeation of the active components through the skin barriers. This external one may also have other active adjuvant substances with different actions on the skin.

[073] The inner layer containing core coating polymer consists of a synthetic lipophilic polymer or combination of more than one synthetic lipophilic polymer (s).

[074] In a different embodiment of the invention, the intermediate layer can also be prepared by conjugating one or more natural hydrophilic polymer (s) and one or more synthetic hydrophilic polymer (s) ). With auxiliary surfactant action, such as agents that promote the stabilization of the nucleus and at the same time promoting the dispersion of the nucleus in the outer layer, they should appear in the intermediate layer with percentages ranging from 0.05% to 95% according to their emulsifying functionality to disperse lipophilic agents (core) within the aqueous solution (outer layer). In terms of the proportion of the intermediate layer in the final dermocosmetic composition, the intermediate coating layer of the core must present percentages between 0.05% and 95%, and preferably between 0.05 and 35% in relation to the total weight of the composition.

[075] The composition of the outer layer, on the other hand, consists of excipients, in addition to other dermatological actives with several functionalities, among which we mention: emollients, consistency agents, softeners, hydrants, humectants, spreadability, refreshing, film-forming, nourishing, rejuvenating, cell renewal, exfoliating, promoters

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44/57 elasticity, anti-wrinkles, antioxidants, sensory modifiers, preservatives, preservatives, dyes, essences. The outer layer can be composed of 1 or more of these excipients and dermatological assets.

[076] In terms of percentages of pharmaceutical excipients in the outer layer, these can represent between 0.05% and 100% of the total weight of the outer layer, according to their functionality in the dermocosmetic matrix. In terms of the proportion of the outer layer in the final dermocosmetic composition, the outer layer of the composition must present percentages between 0.01% and 100%, preferably between 1 and 100% in relation to the total weight of the composition.

[077] The dermocosmetic composition presented here must be packed in packaging for substances sensitive to light, with a matte aspect and shades between amber in the dark and with internal photoprotection to light (optional).

[078] Still, other dermatological assets used in the composition can be of different types and in different formulations, with variable compositions between the different types of dermatological assets, there are hydrophilic dermatological assets, as well as lipophilic dermatological assets, which can be natural or synthetic. such as hyaluronic acid®, glycolic extract of arnica®, mango butter®, kelpadelie®, lotus rutina®, DMAE bitartarate®, Algisium C®, hydroxyprolisilane C®, Ginkgo biloba® liposomes, biorusol®, Ceramide IIIA®, VC- PMG®, lipoic acid®, oily vitamin E, PML AE and AC® liposomes, elastocell®, PML coenzyme Q10® liposomes, sensiline®, structurine®, Adenin®, nutriskin®, raffermine®, helioxine®, titanium dioxide micronized®, Neo heliopan E 1000®, Neo Helliopan OS®, Neo heliopan BB®, Neo heliopan MA®, macadamia oil,

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45/57 pentacare®, remoduline®, dermatan sulfate®, sunflower oil®, fucogel®, collagen liposomes®, glycolic peach extract®, glycolic olive extract®, glycolic apricot extract®, maturine®, alistin®, algisium C®, salicylic acid nanospheres®, DSBC®, glycolic extract of passion fruit®, pentaglycan®, glucan E-20®, sunflower oil®, glycolic extract of melissa®, glycolic extract of melissa®, fucogel®, glycolic extract hypericum®, Iris Iso®, seaweed extract®, GPS vectorized trehalose®, hydrating PABC®, retinoic acid®, hydroquinone®, VC-IP®, skin whitening complex®, water-soluble vitamin F, cafeisilane C®, antilipemic Bioex ®, water-soluble UVA / UVB filter®, benzophen-3®, ethylhexylate methoxycinnamate, octocrylene®, isoamyl® methoxycinnamate, octyla® salicylate, methyl® anthranilate, retinol® micro-sponges, prosaja, densiskin®, soybean oil®, Coup Déclat®, biorusol II SCA®, cherry essence®, Trealose®, phytospingosine®, matrixil®, argireline®, tensine® , elastinol®, ascobosilane C®, whitespheres®, DMAE fluid®, allantoin®, topical genistein®, nutripeptides®, liftline®, kinetin L®, cerealmilk Premium®, biomimetric LRF complex®, silica shells®, sebonormine®, Net FS ®, physiogenyl®, fomblin HC / R®, copaíba oil®, cupuaçu butter®, Sk influx®, hydroviton 24®, palmoconcrete fitoconcentral®, clerilys W®, pelemol IN 2®, lactil®, Clariskin II®, methyl paraben, propylparaben, propylene glycol, BHT, ultraviolet nanovitamin A®, phenogard MP®, cerearet 20®, cetearyl alcohol, mineral oil, lanolin and vaseline alcohol, imidazolidinylurea, self-emulsifying and emollient base for O / A creams and lotions.

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- Uses of NPV in dermocosmetic compositions containing them [079] Faced with biological activities of red propolis extract, nanospheres loaded with red propolis extract can be used in a wide range of industrial sectors in the cosmetic, cosmeceutical, hygiene and cleaning sectors .

[080] The use of propolis as an active ingredient in the prevention and treatment of various diseases is already widely described. However, red propolis has a different composition from other propolis already widely described in the state of the art, which indicates varied biological activities.

[081] The dermocosmetic compositions presented here demonstrate leishmanicidal activity against Leíshmanía (V.) brazílíensís carried out through an assay of leishmanicidal activity whose IC50 concentration inhibits 50% of the parasite population in vitro testing. Such activity shows that the red propolis extract and the nanospheres loaded with red propolis extract can be used against leishmaniasis.

[082] Due to their proven antioxidant biological characteristic, nanospheres loaded with red propolis extract and dermocosmetic compositions containing NPV obtained by the proposed processes can be used for: treatment of the skin and attached organs; treatment and rejuvenation of the skin, treatment of inflammatory processes of seborrheic origin; treatment of wounds in the postoperative period, treatment of decubitus wounds in patients with diabetic foot; treatment of

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47/57 inflammatory processes of the skin in general. Preferably, for the preparation of non-ionic cream containing NPV, suspensions of polymeric nanoparticles loaded with red propolis extract, as well as the solid NPV obtained by lyophilization will be used for the preparation of antioxidant regenerating cream, anti-edema creams, eye area cream , rejuvenating cream, depigmenting cream, skin whitening cream, cream for sagging eye area, anti-cellulite cream, massage cream, Oil-Free anti-signs cream, cream for dry skin, cream for pre-surgery, Post-Surgery cream, anti-aging cream, including for sensitive skin, tired skin, mature skin, cream for eye bags; Photoprotective lotion SPF between 5 to 60, accelerating tanning lotion SPF 10, Moisturizing PostSol lotion, moisturizing body lotion, Post Peeling lotion, depigmenting lotion, Anti-wrinkle lotion with retinol, After-Sun Cream with formononetine, Gel- After-sun cream with isoflavone concentrate, Anti-aging gel with Botox-like action, Nourishing cream-gel, After-bath lotion, Gel for oily skin, Anti-acne gel and oil, Regenerating hand cream, Barrier hand cream , Hand lotion with sunscreen. Such uses for the product proposed here, in terms of the application of polymeric nanoparticles containing red propolis extract, present themselves as a differential compared to what has already been exposed by the state of the art.

[083] Thus, the modalities of the invention described in this document are presented as an advance in the state of the art since they allow the production of NPV and dermocosmetic compositions containing them economically

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48/57 feasible. In this way, the NPV obtained through the proposed process can be used, alone or in composition with other products, in several industrial sectors, such as cosmetics, hygiene and cleaning and cosmeceuticals in the treatment of leishmaniasis.

EXAMPLES [084] The NPV evaluations obtained using the proposed process demonstrated the obtainment of a nanoparticulate product, with nanometric particle size, adequate encapsulation degree and preserved antioxidant activity. The results obtained are shown in the indicated tables and figures.

[085] Table 1 presents the results of mean particle diameter, Polydispersion Index, Zeta Potential and pH of the Nanoparticle Suspensions containing red propolis extract.

Table 1

Compositions Average particle diameter (nm) Polydispersity Index (PDI) Potential Zeta (mV) pH NEUPlacebo 177.6 ± 0.7 0.13 + 54.6 ± 0.80 6.0 ± 0.05 NEU12% 107.1 ± 0.7 0.18 + 37.3 ± 1.30 5.98 ± 0.1 NEU20% 117.4 ± 0.9 0.15 + 40.4 ± 0.60 5.88 ± 0.2 NEU25% 140.4 ± 1.7 0.14 + 44.8 ± 0.50 5.36 ± 0.2 NEU30% 151.9 ± 2.3 0.17 + 37.9 ± 0.90 5.9 ± 0.07 NEU50% - - - 5.6 ± 0.10 NEU70% - - - 5.2 ± 0.10 NPCLPlacebo 279, 6 ± 1.4 0.17 -33.5 ± 6.10 5.05 ± 0.1 NPCL25% 262.2 ± 7.1 0.13 -18.7 ± 0.51 4.53 ± 0.1

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NPCL30% 280, 2 ± 6, 7 0, 17 -26, 8 ± 4, 60 4, 64 ± 0, 1 NPCL50% 208, 5 ± 4, 8 0, 12 -12, 7 ± 5, 30 4, 54 ± 0, 2 NPCL70% 246, 7 ± 7, 0 0, 11 1— ¹ vo 2 ± 4, 60 4, 16 ± 0, 1 NEUPCLPlacebo 221, 1 ± 1, 2 0, 10 + 32, 4 ± 0, 80 5, 78 ± 0, 1 NEUPCL20% 200, 8 ± 1, 6 0, 12 +25, 0 ± 0, 60 6, 58 ± 0, 1 NEUPCL30% 191, 0 ± 1, 8 0, 09 +29, 6 ± 0, 60 6, 30 ± 0, 1 NEUPCL40% 195, 8 ± 3, 4 0, 11 +18, 0 ± 1, 50 5, 65 ± 0, 1

* Values referring to the average of two readings for mean particle diameter and PDI; Values referring to the average of the determination of three values of the zeta potential ± standard deviation; Values referring to the average of the determination of three pH values ± standard deviation.

[086] Particle size values varied between 107.1-177.6nm with polydispersity index between 0.13 to 0.18 for NEU compositions, between 246.7-280.2nm with polydispersion index between 0.11 at 0.17 for NPCL compositions, between 191.0-221.1nm with polydispersity index between 0.09 and 0.12 for NEUPCL compositions and all compositions showed a unimodal behavior. The zeta potential was shown to be between +54.6 to +37.3 mV, for the NEU composition, between -33.5 to -12.7 mV for the NPCL composition and between +32.4 to +18.0 mV for NEUPCL compositions. The compositions proved to be stable, dispersed and with no tendency to aggregate the nanoparticles. The pH values of nanoparticle suspensions containing red propolis extract were similar to the suspension of nanoparticles without red propolis extract (placebo) with slightly acid values between 5.20 to 6.00 for NEU compositions, between 5.05 to 4.16 for NPCL compositions, and between 5.65 to 6.58 for NEUPCL compositions.

[087] The attached Figure 1 shows Electronic Scanning Photomicroscopy of NEU12% (1A, 1B), NEU20% (1C and 1D)

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50/57 and NPCL30% (1E and 1F) without cryoprotectant (1A, 1B and 1D) and with cryoprotectant (1C, 1D and 1F). Photomicrographs with magnification between 3000 times (scale 5pm) and magnification of up to 12000 times (scale 1pm) showing nanometric particles.

[088] The attached Figure 2 shows DSC thermograms of the red propolis extract, the polymeric coating matrix and NPV compositions.

[089] The DSC thermogram of the red propolis extract showed 4 endothermic events at the temperatures of 81.7 ° C, 92.0 ° C, 107 ° C and 135 ° C referring to wax fusion processes and other phenolic components present in red propolis extract, while polymeric coatings containing Eudragit®E-100 (NEU), Poly-εcaprolactone (NPCL) and Eudragit®E-100 (NEU) and Poly-εcaprolactone (NEUPCL) presented an endothermic melting event near temperature 60 ° C and that the NPV compositions were similar in the temperature range between 48 to 59 ° C with a peak in 52 ° C and without additional endothermic events in the temperature range of the red propolis extract, which shows by the thermal properties that the red propolis extract was encapsulated.

[090] The attached Figure 3 shows FTIR-ATR spectra of the red propolis extract, the polymeric coating matrix and NPV compositions.

[091] The FTIR-ATR Spectrum of the red propolis extract showed stretches of phenolic compounds in 3336cm-1 stretches in 1617cm-1, 1496cm-1 and 1450cm-1 (axial strain C = C of aromatic rings in phenolic compounds) and stretches in 1045 cm-1 referring to axial deformation of CO from aromatic ether and 877cm-1 stretches referring to characteristics of (phenolic hydroxyl),

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51/57 angular deformation outside the C-H plane of aromatics. The FTIR-ATR spectra of polymeric coating matrix and NPCL compositions showed similar stretching in 2863cm-1, 1400-1340cm-1 regarding axial and angular deformations, respectively of CH2 and axial and angular deformation of C- (CO) -C in 1171cm-1, in addition to axial deformation of aliphatic ketone carbonyl (C = O) in 1725cm-1, and the disappearance of the characteristic stretches of the red propolis extract proving by encapsulation of the red propolis extract by infrared spectroscopic technique during the process of obtaining of the NPCL.

[092] For compositions featuring Eudragit®E-100 (NEU and NEUPCL) characteristic stretches of Eudragit-E-100 were observed, among them a symmetrical axial strain CH of CH2 and CH3 between 2970-2850 cm-1, an intense stretch ( 1740 cm-1 (1750-1735 cm-1) due to the axial deformation of the C = O ester group, two low intensity absorption bands at 1464 cm-1 and 1378 cm-1 referring to the CH angular deformation of CH2 and CH3 and two bands in 1210 - 1190 cm-1 characteristic of medium-strong axial deformation of COC and C- (C = O) -CO- C ester. Weak-weak axial strain at 1035 cm-1, referring to the low intensity tertiary amine C-N bond. In the NEU and NEUPCL compositions loaded with red propolis extract, suppression of the absorption bands or stretches characteristic of phenolic hydroxyls were observed in the region from 3400 to 3250 cm-1 and the characteristic stretch of aromatic ring in the region from 1600 to 1450 cm-1 in compositions containing these polymeric matrices.

[093] The attached Figure 4 shows the chromatograms (UPLC-DAD) of the red propolis extract (4A), from the matrix

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52/57 coating (4B) (Placebo), NEU30% composition (4C), NPCL30% composition (4D) and NEUPCL30% composition (4E) showing the identification of flavonoids / isoflavonoids present in the propolis extract and NPV compositions and absence of flavonoids in the polymeric coating matrix (placebo).

[094] Figures 4A, 4C, 4D and 4E show chromatograms of analytical runs obtained using a chromatographic method developed to detect the presence of flavonoids. Figures 4A, 4C, 4D and 4E show the presence of flavonoids (1) Liquiritigenina (12.53min.), (2) pinobanksina (15.68min.), (3) isoliquiritigenin (17.26min.), (4) formononetine (18,13min.), (5) pinocembrine (23,12min.) and (6) Biochanin A (23,81min.) in the retention times corresponding and similar to the analytical standards used for the development of the chromatographic method. [095] Table 2 presents the results of Degree of

Encapsulation of major major flavonoids

gifts UPLC-DAD. in the extract of red propolis and in NPV using Table 2 (%) Degree encapsulation of flavonoids UPLC-DAD * majority using Composition A B Ç D AND NEU12% 30.29 ± 2.69 32.12 ± 1.13 42.70 ± 0.14 33.21 ± 0.06 60.16 ± 0.16 NEU20% 20.51 ± 0.57 25.71 ± 0.35 35.70 ± 0.42 27.85 ± 0.74 59.39 ± 0.32 NEU25% 15.52 ± 0.12 20.98 ± 0.13 28.31 ± 0.28 23.12 ± 0.29 52.57 ± 5.48 NEU30% 37.54 ± 1.70 47.55 ± 1.17 62.98 ± 0.03 51.07 ± 0.02 77.58 ± 1.33 NEU50% 24.32 ± 0.09 28.05 ± 0.61 37.39 ± 0.11 32.54 ± 0.05 64.78 ± 0.37 NEU70% 26.34 ± 0.87 29.97 ± 0.03 48.74 ± 0.08 40.97 ± 0.01 72.04 ± 0.80

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NPCL25% 18.41 ± 0.72 31.19 ± 1.11 81.87 ± 1.34 53, 39 ± 0, 85 74, 54 ± 0, 26 NPCL30% 30.71 ± 1.20 39.78 ± 0.21 111.86 ± 0.31 88, 48 ± 0, 02 112 , 79 ± 0 , 7 NPCL40% 27.16 ± 0.36 41.35 ± 0.67 94.01 ± 0.47 78, 67 ± 0, 18 92, 53 ± 0, 86 NPCL70% 27.36 ± 4.45 30.93 ± 0.83 69.94 ± 1.23 60, 58 ± 1, 08 74, 48 ± 0, 10 NEUPCL20 O O 27.22 ± 1.12 43.25 ± 0.35 102.96 ± 1.5 77, 26 ± 0, 64 92, 52 ± 1, 08 NEUPCL30 O O 29.88 ± 0.02 57.63 ± 5.97 106.71 ± 1.2 88, 67 ± 1, 63 94, 87 ± 2, 04 NEUPCL40 O O 29.80 ± 1.01 37.31 ± 0.56 97.92 ± 0.44 78, 02 ± 1, 73 93, 54 ± 0, 77

A: Daidzein; B: Liquiritigenin; C: Isoliquiritigenin; D: Formononetine; E: Biochanina A. * Standard deviation values determined in two chromatographic runs.

[096] Among the studied compositions NEU that presented Eudragit®E-100 was the one that presented the greatest losses during the washing, centrifugation and drying stage using lyophilization technique with encapsulation degree below 60%. For the compositions NPCL and NEUPCL, higher degree of encapsulation was observed in the range between 50% to 77%. It was observed that liquiritigenin and daidzein were the flavonoids that presented the highest losses during the process of obtaining solid NPV. Among the studied compositions, NEU, NPCL and NEUPCL with 30% of red propolis extract presented higher values of content and% of encapsulation (Table 3).

[097] Table 3 presents the results of percentages

of (%) Degree in Encapsulation and (o) Non-encapsulated representing the main major flavonoids present in extract of propolis red and NPV using UPLC-DAD. Table 3 (%) Degree of encapsulation using UPLC-DAD Composition % GE Average of flavonoids NEU12% 39.69 NEU20% 33.83

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NEU25% 28.04 NEU30% 55.34 NEU50% 37.42 NEU70% 43.61 NPCL25% 51.88 NPCL30% 76.72 NPCL40% 66.74 NPCL70% 52.66 NEUPCL20% 68.64 NEUPCL30% 75.43 NEUPCL40% 67.32 GE: Degree of encapsulation; [098] The NPV presented good antioxidant activity and it was concentration dependent. In the concentrations of

red propolis of 80 and 10pg / mL, NPV's showed antioxidant activity between 71.14 to 98.41% and in concentrations of 5 and 2.5pg / mL, NPV compositions showed percentage of antioxidant activity between 58.87 and 97 , 33% and, therefore, much higher than the red propolis extract between 25.97 and 40.73%. These results are satisfactory to demonstrate that the NPV compositions will act as agents that inhibit oxidative processes and functioning as antioxidants at the biological level, such as skin and attached organs, and may function as an anti-aging agent on the skin (Table 4).

[099] Table 4 shows the results of antioxidant activity (%) using DPPH method of red propolis extract and NPV.

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Table 4

Compo % Antioxidant Activity sition 80qg / mL 10qg / mL 5qg / mL 2, 5qg / mL EPV 98.06 ± 0.18 79.00 ± 0.13 40.73 ± 0.03 25, 97 ± 0.04 NCEU 12% 97.30 ± 0.17 71.14 ± 0.06 65.89 ± 0.06 58, 87 ± 0.11 NCEU 20% 97.30 ± 0.42 78.34 ± 0.06 72.17 ± 0.11 67, 07 ± 0.19 NCEU 25% 95, 90 ± 0.17 76.68 ± 0.22 69.77 ± 0.23 66, 70 ± 0.46 NCEU 30% 95.87 ± 0.11 96.12 ± 0.22 77.38 ± 0.22 69, 70 ± 0.06 NCEU 50% 98.37 ± 0.06 97.33 ± 0.25 75.42 ± 0.32 66, 11 ± 0.26 NCEU 70% 100.01 ± 0.11 98.41 ± 0.06 79.67 ± 0.17 68, 96 ± 0.29 NCPCL 20% 99.58 ± 0.55 93.12 ± 0.87 86.07 ± 0.51 80, 12 ± 0.06 NCPCL 30% 97.39 ± 0.36 84.61 ± 0.14 97.39 ± 0.44 86, 18 ± 0.34 NCPCL 40% 94.48 ± 0.37 78.47 ± 0.11 89.52 ± 0.46 86, 09 ± 0.36 NCPCL 70% 94.48 ± 0.05 90.08 ± 0.11 84.11 ± 0.17 76, 50 ± 0.67 NCEUP 20% 100.83 ± 0.46 91.94 ± 0.67 92.30 ± 0.80 74, 31 ± 0.34 NCEUP 30% 101.80 ± 0.97 84.11 ± 0.05 78.52 ± 0.96 72, 87 ± 0.11 NCEUP 40% 101.75 ± 0.60 79.82 ± 0.19 79.79 ± 0.28 75, 33 ± 0.13

* Values referring to the average of the determination of three pH values ± standard deviation.

[100] It is observed that the formulations showed a higher percentage of antioxidant activity than the extract in the free form, showing that these compositions can act for a longer time or in a lower concentration than the bioactive extract of red propolis, increasing the antioxidant power of phenolic compounds . The exacerbation of antioxidant activity in the formulations is probably due to the effect of controlled release of these coating matrices from these matrix systems.

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56/57 [101] The attached Figure 5 presents the determination of the IC50 of the leishmanicidal activity test against Leíshmanía (V.) brazílíensís using normalized graph (figure 5A) and some hypothetical biochemical mechanisms of cell death for Leíshmanía (V.) brazílíensís by flavonoids, especially isoliquiritigenin (chalcone), present in the red propolis extract and in the nanospheres loaded with red propolis extract (figure 5B). The EPV and the nanoparticles of the compositions NEU30% and NPCL30% showed proven leishmanicidal activity with IC50 values of 179.9pg / mL and 31.16 pg / mL, respectively. The compositions NCPCL 30% and NPCL40% (47.23 pg / mL) showed results close to EPV in free form with an IC50 of 38.3pg / mL.

[102] Figure 5 is based on research by Torres-Santos and Chen. Torres-Santos et al. (2009) (Torres-Santos EC, Sampaio-Santos MI, Buckner FS, Yokoyama K, Gelb M, Urbina JA, Rossi-Bergmann B (2009) Altered sterol profile induced in Leishmania amazonensis by a natural dihydroxymethoxylated chalcone. JAntimicrobial Chemother 63: 469-472) demonstrated that chalcones (dihydro-methoxylated chalcone) alter the steroid biosynthesis in L. amazonensís and promote the accumulation of ergosterol and lower cholesterol. This change results in an increase in the fluidity of the membrane and the toxic effect on the parasite with IC 50 values equal to 5.5 μΜ. A study by Chen et al. (2001) (Chen M, Zhai L, Christensen SB, Theander TG, Kharazmi A (2001) Inhibition of fumarate reductase in leishmania major and L. donovani by chalcones. Antimicrobial Agents Chemother 45 (7): 2023-2029) showed that chalcones (licochalcona A in specific study) destroy the ultrastructure of the parasite's mitochondria, inhibit mitochondrial respiration and enzyme activity

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57/57 mitochondrial dehydrogenase, which are succinate dehydrogenase (SDH), succinate cytochrome and reductase, NADH dehydrogenase, NADH cytochrome and reductase, especially the activity of the enzyme fumarate reductase (FDH) present only in the Leíshmanía parasite.

[103] The attached Figure 6 shows photos of nonionic creams containing NPCL30% loaded with red propolis extract to obtain a final concentration in the cosmetic cream of 2.0% (A), 1.5% (B), 1.0% ( C) and 0.5% (D) of the red propolis extract. The solid-state NPV nanospheres were either directly incorporated into the non-ionic cream or using polymeric nanoparticle suspensions (B).

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Claims (50)

Claims 1. Nanospheres loaded with red propolis extract characterized by comprising active principles derived from red propolis. 2. Nanospheres loaded with propolis extract characterized by the active ingredients being derived from red propolis extracts and presenting a final percentage of red propolis extract between 14 and 69%. 3. Nanospheres loaded with red propolis extract, according to claims 1 and 2, characterized by the fact that it comprises: a) standardized extract of red propolis from fresh red propolis from the mangroves of Alagoas in a proportion between 5 and 100%; b) nano-coated encapsulant in a proportion between 5 and 95%, preferably between 30 and 86%; c) dispersant with stabilizing function in a proportion between 5 and 95%, preferably between 15 and 35%. 4. Nanospheres loaded with red propolis extract, according to claim 3, characterized by comprising: a) nano-coating encapsulating system consisting of a binary system, consisting of a) poly-s-caprolactone (PM 10,000) and pluronic F-108 (PM 14,000), or b) poly-scaprolactone (PM 10,000) and Pluronic F-68 for obtain NPCL compositions; Petition 870160041932, of 08/04/2016, p. 68/89 2/13 b) nano-coating encapsulating system composed by binary system, composed by eudragit E-100 (P.M 14.000) and pluronic F-108 (P.M. 14.000) to obtain NEU compositions; c) nano-coating encapsulating system composed of a ternary system, composed of poly-s-caprolactone (P.M 10,000), eudragit E-100 and pluronic F-108 (P.M. 14,000) to obtain NEUPCL compositions. 5. Nanospheres loaded with red propolis extract, according to claims 1 to 4, characterized by having controlled release properties. 6. Nanospheres loaded with red propolis extract, according to claims 1 to 4, characterized by having properties to protect the dermocosmetic composition from chemical incompatibilities between the phenolic compounds present in the propolis extract and other cosmetic excipients. 7. Nanospheres loaded with red propolis extract, according to claims 1 to 4, characterized by having photoprotective properties of the phenolic compounds present in the propolis extract. 8. Nanospheres loaded with red propolis extract, according to claims 1 to 4, characterized by having biocompatibility and biodegradability properties without presenting any risk when applied topically to the skin. 9. Nanospheres loaded with red propolis extract, according to claims 1 to 8, characterized by containing, Petition 870160041932, of 08/04/2016, p. 69/89 3/13 among its components, compounds derived from red propolis, preferably standardized extract of red propolis, for use in dermocosmetic treatment with leishmanicidal action for the treatment of cutaneous leishmaniasis. 10. Nanospheres loaded with red propolis extract, according to claim 9, characterized by containing, among its components, compounds derived from red propolis, preferably standardized red propolis extract, for use in dermocosmetic treatment with leishmanicidal action for the treatment of leishmaniasis and, specifically , caused by the genus Leíshmanía brazílísensís. 11. Nanospheres loaded with red propolis extract, according to claims 9 and 10, characterized by containing, among its components, compounds derived from red propolis, preferably standardized red propolis extract, for use in dermocosmetic treatment with antioxidant and anti-inflammatory action for treatment of cutaneous leishmaniasis caused by Leíshmanía brazílísensís. 12. Nanospheres loaded with red propolis extract, according to claims 1 to 8, characterized by containing, among its components, compounds derived from red propolis, preferably standardized red propolis extract, for use in dermocosmetic treatment with antioxidant and hydrating action in processes erythematous inflammatory, preferably caused by excessive exposure to sunlight or artificial light. 13. Nanospheres loaded with red propolis extract, according to claims 1 to 8, characterized by containing, Petition 870160041932, of 08/04/2016, p. 70/89 4/13 among its components, compounds derived from red propolis, preferably standardized extract of red propolis, for use in dermocosmetic treatment with antioxidant and hydrating action in inflammatory processes of atopic dermatitis. 14. Nanospheres loaded with red propolis extract, according to claims 1 to 8, characterized by containing, among its components, compounds derived from red propolis, preferably standardized red propolis extract, for use in dermocosmetic treatment with antioxidant and anti-wrinkle processes natural skin aging caused, preferably, by exposure of the skin to climatic variations or exposure of the skin to chemical, physical agents, as well as biochemical or nutritional deficiencies. 15. Nanospheres loaded with red propolis extract, according to claims 1 to 8, characterized by containing, among its components, compounds derived from red propolis, preferably standardized red propolis extract, for use in dermocosmetic treatment with antioxidant action delaying premature aging skin and other effects caused by the action of extrinsic agents. 16. Process for obtaining nanospheres loaded with red propolis extract, characterized by comprising the following steps: i) Preparation of the standardized red propolis extract from fresh red propolis; Petition 870160041932, of 08/04/2016, p. 71/89 5/13 ii) Preparation of suspensions of nanospheres loaded with red propolis extract by solubilizing the components of the organic phase and components of the aqueous phase in specific volumes and concentrations, in separate containers, using an ultrasonic bath; iii) Preparation of suspensions of nanospheres loaded with red propolis extract by nanoprecipitation technique of the components of the organic phase over the aqueous phase using specific proportions and concentrations of the components of the organic phase and the aqueous phase, as described in ii); iv) Preparation of the nanospheres loaded with red propolis extract by centrifugation, using specific rotation speed and time followed by the concentration technique of the nanosphere suspensions loaded with red propolis extract; v) Drying the nanospheres loaded with red propolis extract with those obtained in iv) using rapid freezing techniques, followed by freeze drying using low pressure conditions; vi) Characterization of nanosphere suspensions loaded with red propolis extract using techniques of: distribution of the average particle diameter, polydispersity index, zeta potential and pH; vii) Characterization of solid nanospheres loaded with red propolis extract using techniques such as: DSC, FTIR, MEV, UPLC-DAD, antioxidant activity; Petition 870160041932, of 08/04/2016, p. 72/89 6/13 viii) Biological characterization of solid nanospheres loaded with red propolis extract using in vitro method of leishmanicidal activity against Leíshmanía brazílíensís. 17. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step iii), the crude extract is solubilized together with a poly-εcaprolactone encapsulating agent responsible for the nanoencapsulation of the red propolis extract to obtain NPCL compositions. 18. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step iii), the crude extract is solubilized together with an encapsulating agent Eudragit E-100 responsible for the nanoencapsulation of the red propolis extract for obtain NEU compositions. 19. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step iii), the crude extract is solubilized together with poly-εcaprolactone and Eudragit E-100 encapsulating agents responsible for the nanoencapsulation of the extract of red propolis to obtain NEUPCL compositions. 20. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step iii), the dispersing / stabilizing agent pluronic F108 or pluronic F-68, preferably pluronic F-, is added 108 Petition 870160041932, of 08/04/2016, p. 73/89 7/13 triblock copolymer responsible for nanodispersion and stabilization of polymeric nanoparticles in suspension. 21. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step iii) the addition of components of the composition with a proportion of solids between 5% to 80% of the standardized extract of red propolis, between 10% to 70% encapsulating agent, and between 10% to 35% dispersing agent. 22. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step v), the drying of the nanospheres in the solid state can occur in the presence of aerosil cryoprotectants between 0.1 to 10%, as well as sodium starch glycolate between 0.1 to 30%. 23. Process of obtaining suspensions of nanoparticles loaded with red propolis extract, according to claim 16, characterized by the fact that, in step vi) obtain an average particle diameter between 107 to 177 nm, polydispersity index between 0.130 to 0.180, and potential zeta between +37.3 to + 54.6mV for the so-called NEU compositions. 24. Process for obtaining suspensions of nanoparticles loaded with red propolis extract, according to claim 16, characterized by the fact that, in step vi) obtain an average particle diameter between 191 to 221.1 nm, polydispersity index between 0.090 to 0.120, and zeta potential between +18.0 to + 32.4mV for the so-called NEUPCL compositions; Petition 870160041932, of 08/04/2016, p. 74/89 8/13 25. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step vii) the coating of polymeric nanoparticles containing red propolis extract using DSC, FTIR and SEM is verified. 26. Process for obtaining nanospheres loaded with red propolis extract, according to claim 16, characterized by the fact that, in step vii) the degree of encapsulation of the nanospheres loaded with red propolis extract using UPLC-DAD is proved. 27. Process of obtaining suspensions of nanoparticles loaded with red propolis extract, according to claim 16, characterized by the fact that, in step viii) the nanospheres loaded with red propolis extract demonstrate leishmaniciated activity for the compositions NEU and NPCL with values of IC50 of 179.9pg / mL and 31.16 pg / mL, respectively. 28. Composition, characterized by comprising nanospheres loaded with red propolis extract as an active ingredient, alone or in conjunction with other actives. 29. Composition, according to claim 28, characterized by being preferably semi-solid formulations, such as non-ionic cream, gel, gel-cream, lotions, non-ionic lotions, cationic creams, cream ointment, powders, makeup powders, whitening skin, face masks; 30. Composition according to claim 28, characterized in that it comprises (i) a core containing active substance (s) of red propolis combined with coating agent; (ii) Petition 870160041932, of 08/04/2016, p. 75/89 9/13 an intermediate layer of dispersant and stabilizer of the active substance (s) contained in the core; (iii) an oil-aqueous outer layer responsible for the dissolution and permeation processes of the active constituents of the red propolis extract through the skin barriers. 31. Composition, according to claim 28, characterized by having a modified release poly-multifunctional characteristic, antioxidant, leishmanicidal action and photoprotection. 32. Composition, according to claim 28, characterized by having a core i) with the function of protecting the composition against auto-oxidation and photodegradation of the constituents present in the dermocosmetic composition. 33. Composition, according to claim 28, characterized by comprising a core (i) composed of the active substances of red propolis, derived from the standardized extract of red propolis and matrix of biocompatible and biodegradable polymeric coating and, presenting a proportion between 0.01% and 10 %, preferably between 0.01 and 5%, in relation to the total weight of the composition. 34. Composition, according to claim 28, characterized by comprising an intermediate layer (ii) composed of cosmetic excipients, preferably dispersing agents and core stabilizers, in the percentage between 0.01% and 95% of the total weight of the intermediate layer, according to the functionality of the cosmetic excipient in the dermocosmetic matrix and, present a proportion between 0.01% and 100%, preferably between 0.01 and 75%, in relation to the total weight of the composition. Petition 870160041932, of 08/04/2016, p. 76/89 10/13 35. Composition, according to claim 28, characterized by comprising an outer layer (iii) with cosmetic excipients, with several functionalities, preferably by diluting, dispersing and stabilizing the intermediate layer and the core in the composition, as well as by dissolving and permeating the active constituents of red propolis extract by the skin barriers, in the percentage between 0.01% and 100% of the total weight of the outer layer, according to the functionality of the pharmaceutical excipient in the pharmaceutical matrix and, present a proportion between 0.01% and 100% , preferably between 0.01 and 99.9%, in relation to the total weight of the composition. 36. Composition, according to claim 28, characterized in that it comprises in the outer layer (iii) cosmetic excipients, with several functionalities, preferably emollients, consistency agents, softeners, moisturizers, humectants, spreaders, refreshers, film-forming, nutritious, rejuvenating , cell renewal promoters, exfoliants, elasticity promoters, anti-wrinkle action agents, antioxidant action agents, sensory modifiers, preservatives, preservatives, odorizers and, present a proportion between 0.01% and 100%, preferably between 0.01 and 99 , 9%, in relation to the total weight of the composition. 37. Composition according to claim 28, characterized by comprising an outer layer with cosmetic excipients such as emollients, consistency agents, moisturizers, moisturizers, humectants, spreadability, refreshing, film-forming, nourishing, rejuvenating, cell renewal, exfoliating, elasticity promoters , anti-wrinkles, antioxidants, sensory modifiers, Petition 870160041932, of 08/04/2016, p. 77/89 11/13 preservatives, preservatives, odorizers and, present a proportion between 0.01% and 100%, preferably between 0.01 and 99.9%, in relation to the total weight of the composition. 38. Composition, according to claim 28, characterized by comprising an outer layer with and other hydrophilic and lipophilic dermatological assets, which may be natural or synthetic and have a proportion between 0.01% and 100%, preferably between 0.01 and 99.9% , in relation to the total weight of the composition. 39. Use of compositions combined with nanospheres loaded with red propolis extract, according to claims 9, 10, 11 and 27, characterized by being a leishmanicidal agent alone or as an adjuvant, to treat cutaneous leishmaniasis caused by the parasite Leíshmanía sp. 40. Use of compositions combined with nanospheres loaded with red propolis extract, according to claims 9, 10, 11 and 27, characterized by being either a leishmanicidal agent or as an adjuvant, to combat skin lesions caused by Leíshmanía (V.) braziliensis. 41. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or as an adjuvant agent, for dermocosmetic treatment with antioxidant action, healing of wounds caused by cutaneous leishmaniasis. 42. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being isolated Petition 870160041932, of 08/04/2016, p. 78/89 12/13 or as an adjuvant agent, for dermocosmetic treatment with antioxidant, healing action in pre-surgical and post-surgical treatment of wounds. 43. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or as an adjuvant, for dermocosmetic treatment with antioxidant, nourishing and aesthetic action of the skin caused by biochemical or nutritional deficiencies. 44. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being an antioxidant agent or as an adjuvant, to combat anti-aging processes of the skin and attached organs. 45. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or with an adjuvant agent, for dermocosmetic treatment of skin cell renewal, anti-inflammatory and moisturizing processes in erythematous inflammatory processes caused by overexposure to sunlight or artificial light. 46. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or as an adjuvant, for dermocosmetic treatment with antioxidant and anti-wrinkle action in natural skin aging processes caused by exposure of the skin Petition 870160041932, of 08/04/2016, p. 79/89 13/13 skin to climatic variations or skin exposure to chemical and physical agents. 47. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or as an adjuvant, for dermocosmetic treatment with antioxidant action and in the lipoxidation of localized fats and cellulite. 48. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or as an adjuvant, for dermocosmetic treatment with anti-acne action and for the treatment of skins with excessive oil production. 49. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being either alone or as an adjuvant agent, for dermocosmetic and aesthetic skin lightening treatment. 50. Use of compositions combined with polymeric nanoparticles containing red propolis extract, according to claims 28 to 38, characterized by being alone or as an adjuvant agent, for dermocosmetic and aesthetic treatment of skin hypopigmentation and treatment of vitiligo. Petition 870160041932, of 08/04/2016, p. 80/89 1/6 DRAWINGS Petition 870160041932, of 08/04/2016, p. 81/89 2/6