Existing methods of treating glioma are not effective for eradicating the disease. Therefore, new and innovative methods of treatment alone or in combination with existing therapies are necessary. Delivery of therapeutic agents through delivery carriers such as liposomes diminishes the harmful effects of the agent in healthy tissues and allows increased accumulation in the tumor. In addition, targeted chemotherapy using liposomes provides the opportunity for further increase in drug accumulation in tumor. However, the current targeting strategies suffer accelerated plasma clearance and are not advantageous in improving efficacy. The search for new tumor targets, novel ligands, new strategies for targeting, and particle stabilization will advance our ability to improve delivery at the tumor level while decreasing toxicity to normal tissues. The global objective of this thesis was to improve the status of current liposomal therapy to achieve higher efficacy in tumors. Here, we show a novel mechanism to increase targeting to tumor while uncompromising on the long circulation of stealth liposomes. Long circulation is essential for passive accumulation of the nanocarriers due to EPR effect, in order to see benefits of targeting. Using phage display technique, a variety of tumor specific peptides were identified for use as targeting moieties. One potential advantage of the approach proposed here is the rapid identification of patient tumor specific peptide that evades the RES. This could lead to the development of a nanocarrier system with high avidity and selectivity for tumors. Therefore, tumor accumulation of the targeted formulations will be higher than that of non‐targeted liposomes due to increased drug retention at the tumor site and uncompromised blood residence time.In addition, it has been shown that the distribution of nanocarriers, spatially within the tumor, is limited that might further hinder the distribution of the encapsulated drug, thereby limiting efficacy. It is necessary to release the drug from within the nanocarrier to promote increased efficacy. Here, we were able to address the problem of drug diffusion within the tumor interstitium using a combination therapy employing a remotely triggered thermosensitive liposomal chemotherapeutic. We fabricated a thermosensitive liposomal nanocarrier that maintained its stability at physiological temperature to minimize toxicity to healthy cells. We, then, showed a remote triggering mechanism mediated by gold nanorods heated via NIR can help in achieving precise control over the desired site for drug release. These strategies enabled increased drug availability at the tumor site and contributed to tumor retardation. Additionally, we show that the synergistic therapy employing gold nanorods and thermosensitive liposomes may have great potential to be translated to the clinic.

L'ARN interférence est un mécanisme d'inhibition post-transcriptionnel, capable de réguler l'expression des gènes. Ce mécanisme endogène, activé par l'intermédiaire de microARN, peut être détourné après transfection de cours fragments d'ARN synthétiques, notamment les siARN. Cette technique autorise ainsi le ciblage spécifique de l'ensemble des gènes composant le génome, dont l'extinction transitoire permet d'étudier à la fois leurs fonctions, mais aussi de découvrir de nouvelles cibles thérapeutiques ou de nouveaux biomarqueurs. Ce très fort potentiel pour la recherche in vitro se retrouve également in vivo, où l'ARN interférence peut être directement utilisé comme agent thérapeutique pour des situations pathologiques telles que les cancers, les infections ou les maladies systémiques. Cependant, la délivrance intra-cytoplasmique des ARN interférents exogènes est nécessaire pour déclencher ce mécanisme de régulation. À l'heure actuelle, en dépit de nombreuses méthodes de transfection développées dans la littérature, cette étape de délivrance reste une limite importante selon les applications envisagées.En ce sens, ces travaux de thèse ont permis de développer un nouveau vecteur à base de nanoparticules lipidiques cationiques, les cLNP, dédié à la transfection cellulaire de siARN. Cette formulation de cLNP a été adaptée, à l'aide d'un plan d'expérience, d'une formulation neutre de LNP permettant l'encapsulation de molécules lipophiles pour des applications en imagerie de fluorescence et/ou de délivrance de médicaments liposolubles. Les caractérisations physico-chimiques des particules cLNP ont démontré une très forte stabilité colloïdale, à la fois pour dans les tampons aqueux et dans les milieux de culture cellulaire complémentés par du sérum. En outre, ces nano-vecteurs se sont avérés extrêmement efficaces pour établir et conserver des liaisons électrostatiques avec des siARN, permettant ainsi d'obtenir rapidement des complexes démontrant une stabilité élevée dans le temps. Les efficacités d'inhibition fonctionnelle de ces nanoparticules ont été testées avec succès sur 3 lignées cellulaires différentes (PC3, HeLa et U2OS). L'ensemble des résultats obtenus confirme le fort potentiel de ce nouveau nano-vecteur, en termes d'inhibition fonctionnelle et d'absence de cytotoxicité, et le positionne parmi les meilleurs agents de transfection commerciaux testés. Ces caractéristiques sont complétées par des capacités de multi-modalité, dont la possibilité d'encapsuler dans le cœur des particules des drogues ou des fluorophores lipophiles. Enfin, des tests préliminaires réalisés sur des cellules considérées comme difficile à transfecter (cellules primaires, cellules non-adhérentes, neurones), ou sur des structures cellulaires tridimensionnelles plus complexes, ouvrent de nouvelles perspectives extrêmement prometteuses. / L'auteur n'a pas fourni de résumé en anglais

La résistance aux médicaments constitue un obstacle majeur pour le traitement du cancer. Les systèmes nanoparticulaires de délivrance de médicaments (nanoparticule drug delivery system, NDDS) sont pressentis pour apporter un nouvel espoir dans le traitement du cancer afin de surmonter la résistance aux médicaments en délivrant spécifiquement l’agent anticancéreux dans la lésion tumorale par effet EPR. Cela aura pour effet d’augmenter la concentration locale en médicaments et par conséquent d’améliorer l'efficacité thérapeutique tout en épargnant les tissus sains afin d'éviter les effets secondaires liés à la thérapie. Dans la mesure où la tumeur a souvent un microenvironnement acide, nous souhaiterions en outre doter nos nanoparticules NDDS d’une sensibilité pH-dépendante afin de permettre une délivrance spécifique dans la tumeur. Au cours de cette thèse, nous avons élaboré différents NDDSs sensibles aux variations de pH en employant des stratégies différentes. Ces NDDSs peuvent spécifiquement libérer le médicament au niveau du tissu tumoral et dans les cellules elles-mêmes à des valeurs de pH acides. En augmentant la concentration intracellulaire de médicament, l'objectif de surmonter la résistance aux médicaments pourrait ainsi être atteint. La présente étude a permis de fournir de nouvelles connaissances sur la conception de nano-transporteurs pour surmonter la résistance multidrogue par l’élaboration de NDDS sensibles au pH et constitue donc un exemple illustrant parfaitement le fait que les progrès des nanotechnologies peuvent être avantageusement mis en œuvre pour développer de nouvelles perspectives thérapeutiques. / Drug resistance presents a great hurdle to cancer treatment. Nanotechnology-based drug delivery systems (NDDSs) are widely expected to bring new hope for cancer therapy to overcome drug resistance by specifically delivering anticancer drugs to tumor lesions via the EPR effect, hence increasing local drug concentrations and consequently enhancing therapeutic efficacy, and at the same time, sparing healthy tissues to avoid side effects. As tumors often have an acidic microenvironment, we would like to further endow the NDDS with a pH-responsive drug releasing property for specific tumor targeting. In this thesis, we established different pH-responsive NDDSs by employing different strategies. These NDDSs could specifically control drug release at tumor tissues and within tumor cells in response to acidic pH. By increasing the intracellular drug concentration, the goal of circumventing drug resistance in cancer was achieved. The present study provides new insights into the design of nanocarriers to overcome drug resistance through pH-responsive drug delivery, and illustrates how advances in nanotechnology can be advantageously implemented to enhance therapeutic outcomes.

Le cancer est l'une des principales causes de décès dans le monde et reste une maladie difficile à traiter du fait des difficultés de pronostic, du développement rapide de métastases et de la résistance aux médicaments. Il en résulte une forte demande en méthodologies d'imagerie innovantes pour le diagnostic précoce et précis ainsi qu’en nouveaux agents anticancéreux possédant de nouveaux mécanismes pour surmonter la résistance aux médicaments. Le but de mon projet de recherche de doctorat était donc de contribuer à cet objectif.La première partie de ma thèse de doctorat a porté sur la création de systèmes sensibles et précis d'imagerie pour la détection de tumeurs cancéreuses en utilisant une nanotechnologie novatrice permettant la délivrance des agents d'imagerie spécifiquement dans les lésions tumorales. Nous avons conçu de nouveaux dendrimères amphiphiles pour assurer le transport de différents agents d'imagerie pour les imageries PET/SPECT, par résonance magnétique et par fluorescence optique. Ces systèmes d'imagerie ont été préparés soit par encapsulation de petites sondes d'imagerie à l'intérieur de nanomicelles dendritiques ou par fonctionnalisation de la surface hydrophile ou de la queue hydrophobe du dendrimère. La deuxième partie a eu pour objectif de développer de nouveaux agents anticancéreux possédant nouveaux mécanismes d’action et une meilleure activité antitumorale. A cet effet, nous avons conçu une série de nucléosides arylvinyltriazoles par réaction oxydante de Heck, ce qui nous a permis d'obtenir les composés désirés pourtant difficiles à synthétiser avec un très large éventail de substrats et une stéréosélectivité unique. / Cancer is one of the leading causes of death in the world, and remains a difficult disease to treat because of poor prognosis, rapid tumor metastasis and drug resistance. Therefore, innovative imaging modalities for early and precise diagnosis as well as new anticancer drug candidates with novel mechanisms to overcome drug resistance are in high demand. The aim of my PhD research project was to contribute to this goal.The first part of my PhD thesis was focused on establishing sensitive and precise imaging systems for cancer detection using innovative nanotechnology to deliver imaging agents specifically into tumor lesions. We designed and constructed novel amphiphilic dendrimers to carry different imaging agents for PET/SPECT imaging, magnetic resonance imaging and optical fluorescence imaging. These innovative imaging systems were prepared by either encapsulation of small imaging probes within the dendrimer nanomicelles, or functionalization of the dendrimer hydrophilic surface or hydrophobic tail. The second part of my PhD program aimed to develop new anticancer drug candidates with novel mechanisms for better anticancer activity. Therefore, we designed and synthesized a series of challenging arylvinyltriazole nucleosides via the oxidative Heck reaction, which allowed us to obtain the desired compounds with excellent substrate scope and unique stereoselectivity.

A Terapia Fotodinâmica (TFD) é uma técnica para tratamento de câncer que usa um fotossensibilizador (FS) na presença de luz e oxigênio molecular gerando espécies altamente reativas de oxigênio que levam as células tumorais à morte. Porém a hidrofobicidade de alguns FSs podem induzir a agregação em sistemas biológicos, com redução da sua atividade fotodinâmica. A incorporação de FSs em sistemas nanocarreadores pode ser uma alternativa para superar este problema. O objetivo deste trabalho foi preparar e caracterizar dois FSs hidrofóbicos (Hipericina e Tetra-carboxiftalocianinade zinco) encapsulados em nanopartículas lipídicas sólidas (NLS) para um potencial uso em terapia fotodinâmica. Os FSs incorporados em nanopartículas lipídicas sólidas foram preparados utilizando a técnica de ultra-sonicação e a caracterização físico-química foi realizada. O tamanho médio das nanopartículas de hipericina e tetra-carboxiftalocianinade zinco foram de 153 e 245 nm respectivamente, índice de polidispersão de 0,28 para Hy-NLS e 0,29 para FtZnT-NLS. Uma das vantagens dos sistemas de encapsulação utilizando NLS é o alto valor de eficiência de encapsulação (EE%) e neste estudo foram obtidos valores de eficiência de encapsulação superior a 80% para a Hy-NLS e FtZnT-NLS. De modo a obter a eficiência fotodinâmica da Hy e FtZnT antes e depois do encapsulamento em NLS, as constantes de velocidade de foto-decomposição utilizando dois agentes captadores de 1O2 (1,3 Difenilisobenzofurano e ácido úrico) foram determinadas. As constantes de velocidade de foto-decomposição tiveram aumento significativo após o encapsulamento que ocorreu provavelmente devido a um aumento no tempo de vida do estado triplete causado pelo aumento da solubilidade. Hy-NLS e FtZnT-NLS apresentaram um aumento acima de 30% e 60% respectivamente na acumulação intracelular e uma melhoria na fototoxicidade correlacionado com o aumento da acumulação intracelular. Todas essas vantagens sugerem que hipericina e a tetra-carboxiftalocianinade zincoencapsuladas em nanopartículas lipídicas sólidas tem potencial para serem utilizadas em terapia fotodinâmica. / Photodynamic therapy (PDT) is a technique for treating cancer using a photosensitizer (PS) in the presence of light and molecular oxygen generating highly reactive oxygen species that lead to tumor cell death. The hydrophobicity of some photosensitizers can induce aggregation in biological systems, reducing its photodynamic activity. The incorporation of PSs in nanocarriers can be an alternative to overcome this problem. The aim of this work was to prepare and characterize two hydrophobic photosensitizers (Hypericin and Zinc tetra-carboxylicphthalocyanine) encapsulated in solid lipid nanoparticles (SLN) for potential use in photodynamic therapy. The PSs incorporated into solid lipid nanoparticles were prepared using the ultrasonication technique, and physico-chemical characterization was performed. The average size of the nanoparticles with hypericin and zinc tetra-carboxylicphthalocyanine was 153 and 245 nm respectively, the polydispersivity index of 0.28 to Hy-SLN and 0.29 to FtZnT-SLN. One of the advantages of encapsulation systems using SLN is the high value of encapsulation efficiency (EE %). In this study were obtained values of encapsulation efficiency greater than 80% for the Hy-SLN and FtZnT-SLN. In order to obtain the photodynamic efficiency of Hy and FtZnT before and after encapsulation in SLN, rate constants using photo-decomposition of two scavengers of 1O2 agents (1,3-Diphenylisobenzofuran and uric acid) were determined. The rate constants of photo-decomposition had significant increase after encapsulation which occurred probably due to an increase in the lifetime of the triplet state caused by the increased solubility. Hy-SLN and FtZnT-SLN showed an increase above 30% and 60% respectively in the intracellular accumulation and an improvement in phototoxicity correlated with increased intracellular accumulation. So, all these advantages suggest that hypericin and zinc tetra-carboxylicphthalocyanine encapsulated in solid lipid nanoparticles have potential to be used in photodynamic therapy.

A Terapia Fotodinâmica (TFD) é uma técnica para tratamento de câncer que usa um fotossensibilizador (FS) na presença de luz e oxigênio molecular gerando espécies altamente reativas de oxigênio que levam as células tumorais à morte. Porém a hidrofobicidade de alguns FSs podem induzir a agregação em sistemas biológicos, com redução da sua atividade fotodinâmica. A incorporação de FSs em sistemas nanocarreadores pode ser uma alternativa para superar este problema. O objetivo deste trabalho foi preparar e caracterizar dois FSs hidrofóbicos (Hipericina e Tetra-carboxiftalocianinade zinco) encapsulados em nanopartículas lipídicas sólidas (NLS) para um potencial uso em terapia fotodinâmica. Os FSs incorporados em nanopartículas lipídicas sólidas foram preparados utilizando a técnica de ultra-sonicação e a caracterização físico-química foi realizada. O tamanho médio das nanopartículas de hipericina e tetra-carboxiftalocianinade zinco foram de 153 e 245 nm respectivamente, índice de polidispersão de 0,28 para Hy-NLS e 0,29 para FtZnT-NLS. Uma das vantagens dos sistemas de encapsulação utilizando NLS é o alto valor de eficiência de encapsulação (EE%) e neste estudo foram obtidos valores de eficiência de encapsulação superior a 80% para a Hy-NLS e FtZnT-NLS. De modo a obter a eficiência fotodinâmica da Hy e FtZnT antes e depois do encapsulamento em NLS, as constantes de velocidade de foto-decomposição utilizando dois agentes captadores de 1O2 (1,3 Difenilisobenzofurano e ácido úrico) foram determinadas. As constantes de velocidade de foto-decomposição tiveram aumento significativo após o encapsulamento que ocorreu provavelmente devido a um aumento no tempo de vida do estado triplete causado pelo aumento da solubilidade. Hy-NLS e FtZnT-NLS apresentaram um aumento acima de 30% e 60% respectivamente na acumulação intracelular e uma melhoria na fototoxicidade correlacionado com o aumento da acumulação intracelular. Todas essas vantagens sugerem que hipericina e a tetra-carboxiftalocianinade zincoencapsuladas em nanopartículas lipídicas sólidas tem potencial para serem utilizadas em terapia fotodinâmica. / Photodynamic therapy (PDT) is a technique for treating cancer using a photosensitizer (PS) in the presence of light and molecular oxygen generating highly reactive oxygen species that lead to tumor cell death. The hydrophobicity of some photosensitizers can induce aggregation in biological systems, reducing its photodynamic activity. The incorporation of PSs in nanocarriers can be an alternative to overcome this problem. The aim of this work was to prepare and characterize two hydrophobic photosensitizers (Hypericin and Zinc tetra-carboxylicphthalocyanine) encapsulated in solid lipid nanoparticles (SLN) for potential use in photodynamic therapy. The PSs incorporated into solid lipid nanoparticles were prepared using the ultrasonication technique, and physico-chemical characterization was performed. The average size of the nanoparticles with hypericin and zinc tetra-carboxylicphthalocyanine was 153 and 245 nm respectively, the polydispersivity index of 0.28 to Hy-SLN and 0.29 to FtZnT-SLN. One of the advantages of encapsulation systems using SLN is the high value of encapsulation efficiency (EE %). In this study were obtained values of encapsulation efficiency greater than 80% for the Hy-SLN and FtZnT-SLN. In order to obtain the photodynamic efficiency of Hy and FtZnT before and after encapsulation in SLN, rate constants using photo-decomposition of two scavengers of 1O2 agents (1,3-Diphenylisobenzofuran and uric acid) were determined. The rate constants of photo-decomposition had significant increase after encapsulation which occurred probably due to an increase in the lifetime of the triplet state caused by the increased solubility. Hy-SLN and FtZnT-SLN showed an increase above 30% and 60% respectively in the intracellular accumulation and an improvement in phototoxicity correlated with increased intracellular accumulation. So, all these advantages suggest that hypericin and zinc tetra-carboxylicphthalocyanine encapsulated in solid lipid nanoparticles have potential to be used in photodynamic therapy.

Made available in DSpace on 2018-06-27T18:56:23Z (GMT). No. of bitstreams: 3 Denise Bom Baldissera.pdf: 939670 bytes, checksum: 68115013c164645744ccf4e19e85a72e (MD5) Denise Bom Baldissera.pdf.txt: 162065 bytes, checksum: 86ebe1a57531232453a3049c98d7c692 (MD5) Denise Bom Baldissera.pdf.jpg: 3681 bytes, checksum: 1841930f64ea683f73bf3a5feed3dc8e (MD5) Previous issue date: 2014-04-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Gynoid lipodystrophy (GLD), also known as cellulite, is a common dermatological condition among women after the puberty, mainly concentrated in the pelvic region, lower limbs and abdomen. It´s a tissue edematous infiltration that promotes unattractive changes, the skin becomes to have a paddled aspect or orange peel , conducing to a low self esteem. The exactly etiology of GLD is unknown, however, its presence and level are related to several factors like genetic, emotional, metabolic and hormonal, besides the age, sex, blood pressure, obesity and habits like smoke, sedentary, tight clothes and poor diet. There are many ways to treat this condition, being the most used and easy accessed the use of cosmetic products having as active principles the vasopressure action, anti-inflammatory, lower microcirculation and lipolytic agents. In the present work, methods for analysis of the lipid nanocarrier containg actives employees for GLD treatment (diidromiricetin, diosgenin and synephrine) were developed and validated. There aren t decrypted monographs in pharmacopeias to analyze these substances in pharmaceutical formulas. In the spectrophotometric method UV-VIS the diosgenin and the dihydromyricetin were quantified in 434 nm and 561 nm, respectively, used as reagents sulfuric acid and ρ- anisaldehyde. Synephrine can be quantified by the derivative ultraviolet spectrophotometry, 239 nm wave-length, in the second order, medium scan, Δλ 10.000 and scale factor of 50, using ethanol as a solvent. The methods showed to be specific, linear, precise, exact and robust to the actives quantification. It can be concluded that the validated methods were of easy execution, fast and with a low cost, to do the quality control analyses. / A lipodistrofia ginoide (LDG), também chamada de celulite, é uma afecção dermatológica comum entre mulheres após a puberdade, concentrando-se preferencialmente na região pélvica, membros inferiores e abdome. Trata-se de uma infiltração edematosa do tecido conjuntivo que promove alterações esteticamente desagradáveis, fazendo com que a pele assuma um aspecto acolchoado ou casca de laranja , levando a uma diminuição da auto-estima. A exata etiologia da LDG é desconhecida, contudo, sua presença e grau de comprometimento estão relacionados a diversos fatores como genéticos, emocionais, metabólicos e hormonais, além da idade, sexo, hipertensão arterial, obesidade e hábitos como fumo, sedentarismo, roupas apertadas e má alimentação. Existem inúmeras formas de se tratar essa condição, sendo a mais utilizada e de fácil acesso o uso de cosméticos contendo princípios ativos com ação vasopressora, anti-inflamatória, estimuladores da microcirculação periférica e agentes lipolíticos. No presente trabalho, métodos para análise de um nanocarreador lipídico contendo ativos empregados para o tratamento da LDG (diidromiricetina, diosgenina e sinefrina) foram desenvolvidos e validados. Não há monografias descritas em farmacopéias para análise desses ativos em formulações farmacêuticas. No método espectrofotométrico UV-VIS a diosgenina e a diidromiricetina foram quantificadas em 434 nm e 561 nm, respectivamente, utilizando como reagentes ácido sulfúrico e ρ-anisaldeido. A sinefrina pode ser quantificada por espectrofotometria ultravioleta derivada, no comprimento de onda de 239 nm, na segunda derivada, velocidade de varredura média, Δλ 10.000 e fator escala de 50, utilizando etanol como solvente. Os métodos demonstraram serem específicos, lineares, precisos, exatos e robustos para a quantificação dos ativos. Sendo assim, pode-se concluir que os métodos validados foram de fácil execução, rápidos e de baixo custo, para a realização das análises de controle de qualidade.

Submitted by MARCIA ROVADOSCHI (marciar@unifra.br) on 2018-08-16T18:45:36Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_DeniseBomBaldissera.pdf: 939659 bytes, checksum: 4e6775a2df22405fe611e164c8aaaba0 (MD5) / Made available in DSpace on 2018-08-16T18:45:36Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_DeniseBomBaldissera.pdf: 939659 bytes, checksum: 4e6775a2df22405fe611e164c8aaaba0 (MD5) Previous issue date: 2014-04-29 / Gynoid lipodystrophy (GLD), also known as cellulite, is a common dermatological condition among women after the puberty, mainly concentrated in the pelvic region, lower limbs and abdomen. It´s a tissue edematous infiltration that promotes unattractive changes, the skin becomes to have a paddled aspect or “orange peel”, conducing to a low self esteem. The exactly etiology of GLD is unknown, however, its presence and level are related to several factors like genetic, emotional, metabolic and hormonal, besides the age, sex, blood pressure, obesity and habits like smoke, sedentary, tight clothes and poor diet. There are many ways to treat this condition, being the most used and easy accessed the use of cosmetic products having as active principles the vasopressure action, anti-inflammatory, lower microcirculation and lipolytic agents. In the present work, methods for analysis of the lipid nanocarrier containg actives employees for GLD treatment (diidromiricetin, diosgenin and synephrine) were developed and validated. There aren’t decrypted monographs in pharmacopeias to analyze these substances in pharmaceutical formulas. In the spectrophotometric method UV-VIS the diosgenin and the dihydromyricetin were quantified in 434 nm and 561 nm, respectively, used as reagents sulfuric acid and ρ- anisaldehyde. Synephrine can be quantified by the derivative ultraviolet spectrophotometry, 239 nm wave-length, in the second order, medium scan, Δλ 10.000 and scale factor of 50, using ethanol as a solvent. The methods showed to be specific, linear, precise, exact and robust to the actives quantification. It can be concluded that the validated methods were of easy execution, fast and with a low cost, to do the quality control analyses. / A lipodistrofia ginoide (LDG), também chamada de celulite, é uma afecção dermatológica comum entre mulheres após a puberdade, concentrando-se preferencialmente na região pélvica, membros inferiores e abdome. Trata-se de uma infiltração edematosa do tecido conjuntivo que promove alterações esteticamente desagradáveis, fazendo com que a pele assuma um aspecto acolchoado ou “casca de laranja”, levando a uma diminuição da auto-estima. A exata etiologia da LDG é desconhecida, contudo, sua presença e grau de comprometimento estão relacionados a diversos fatores como genéticos, emocionais, metabólicos e hormonais, além da idade, sexo, hipertensão arterial, obesidade e hábitos como fumo, sedentarismo, roupas apertadas e má alimentação. Existem inúmeras formas de se tratar essa condição, sendo a mais utilizada e de fácil acesso o uso de cosméticos contendo princípios ativos com ação vasopressora, anti-inflamatória, estimuladores da microcirculação periférica e agentes lipolíticos. No presente trabalho, métodos para análise de um nanocarreador lipídico contendo ativos empregados para o tratamento da LDG (diidromiricetina, diosgenina e sinefrina) foram desenvolvidos e validados. Não há monografias descritas em farmacopéias para análise desses ativos em formulações farmacêuticas. No método espectrofotométrico UV-VIS a diosgenina e a diidromiricetina foram quantificadas em 434 nm e 561 nm, respectivamente, utilizando como reagentes ácido sulfúrico e ρ-anisaldeido. A sinefrina pode ser quantificada por espectrofotometria ultravioleta derivada, no comprimento de onda de 239 nm, na segunda derivada, velocidade de varredura média, Δλ 10.000 e fator escala de 50, utilizando etanol como solvente. Os métodos demonstraram serem específicos, lineares, precisos, exatos e robustos para a quantificação dos ativos. Sendo assim, pode-se concluir que os métodos validados foram de fácil execução, rápidos e de baixo custo, para a realização das análises de controle de qualidade.