Würzburg, Universiẗat, Diss., 2004.

Düsseldorf, Universiẗat, Diss., 2002.

O objetivo desta pesquisa foi o desenvolvimento, avaliação da estabilidade e da eficácia de formulações cosméticas contendo acetil hexapeptídeo-3 (AHP) ou acetil tetrapeptídeo-5 (ATP) ou mistura de di e tripeptídeos (DTP). Para tal, foram desenvolvidas 6 formulações à base hidroxietilcelulose (HEC), metilfenil polissiloxano (MFP) e álcool batílico e lecitina de soja (ABL) (F1), ou de HEC, MFP, ésteres de polietilenoglicol com cadeia alquílica com 12 a 20 carbonos (EP) (F2), ou de polímero de acrilatos (PA), MFP e ABL (F3) ou de PA, MFP e EP (F4), ou de HEC e MFP (F5) ou de PA e MFP (F6), contendo ou não (veículo) 10% de AHP, ou 10% de ATP ou 4% de DTP, as quais foram submetidas a um estudo de estabilidade física por determinação da reologia e a uma avaliação sensorial. Para o estudo de eficácia dos peptídeos objeto de estudo as formulações foram aplicadas nos antebraços, na face, na região dos olhos e da boca de 60 voluntárias, sendo realizadas medidas do conteúdo aquoso do estrato córneo, da perda de água transepidérmica (TEWL), do micro-relevo cutâneo e das propriedades mecânicas da pele (anisotropia), antes (basal), após 2 horas da aplicação única (efeitos imediatos) e após 15 e 30 dias de uso contínuo (efeitos a longo prazo) das formulações. Os dados foram analisados estatisticamente pelo teste paramétrico análise de variância. Nos testes de estabilidade física, as formulações de nos 1, 2 e 5 foram consideradas as mais estáveis e as formulações de nº 5 e 6 foram as que apresentaram melhor sensorial, sendo, portanto, selecionada a formulação de nº 5 como veículo para os estudos de eficácia. Na avaliação dos efeitos imediatos na pele, observou-se que as formulações estudadas proporcionaram um aumento significativo no conteúdo aquoso do estrato córneo dos antebraços, enquanto que, somente as formulações acrescidas dos peptídeos provocaram um aumento significativo neste conteúdo na face. Em relação ao micro-relevo cutâneo, as formulações contendo ou não os peptídeos provocaram uma redução do parâmetro Ser (aspereza da pele) e Rt (rugosidade) nos antebraços e a formulação contendo DTP provocou um efeito imediato significativo no parâmetro Sesm (textura da pele). Na avaliação dos efeitos a longo prazo, as formulações ocasionaram uma diminuição significativa na TEWL e as acrescidas dos peptídeos objetos de estudo proporcionaram um aumento significativo no conteúdo aquoso do estrato córneo na região dos olhos, enquanto que nos antebraços e na região da boca todas as formulações acrescidas ou não dos peptídeos ocasionaram um aumento nesse parâmetro. Em relação ao micro-relevo cutâneo, a aplicação das formulações contendo os peptídeos, na região da boca, provocou um aumento significativo nos valores de Sesm. A aplicação das formulações contendo os peptídeos objeto de estudo, nos antebraços ocasionou uma diminuição significativa da anisotropia da pele. A formulação nº 5 contendo os peptídeos objeto de estudo, apresentou efeitos imediatos na hidratação, aspereza, rugosidade e textura da pele e, efeitos a longo prazo, na hidratação, textura e anisotropia da pele, mostrando a importância do emprego destes peptídeos em produtos cosméticos com finalidades hidratantes e antienvelhecimento. / The aim of this study was the development, evaluation of the stability and the efficacy of cosmetic formulations containing acetyl hexapeptide-3 (AHP), or acetyl tetrapeptide-5 (ATP) or mixture of di- and peptides (DTP). For this purpose the 6 following formulations (F) were developed: F1- containing hydroxyethyl cellulose (HEC), methylphenyl polysiloxane (MPPS), batyl alcohol, and lecithin (BAL); F2- HEC, MPPS and C12-C20 PEG-8 alkyl ester (AE); F3- acrylate copolymer (AC), MPPS and BAL; F4- AC, MPPS and AE; F5- HEC and MPPS and F6- AC and MPPS, which were supplemented or not (vehicle) with 10% of AHP or 10% of ATP or 4% of DTP and submitted to the evaluation of physical stability by the determination of the rheologic behavior and also to the sensorial analysis. For clinical efficacy studies, formulations were applied to the volar forearm and to the face skin (eye corner and cheek region), of 60 female volunteers. Skin conditions in terms of skin moisture, transepidermal water loss (TEWL), skin mechanical properties (anisotropy) and skin microrelief, before and after 2 hours (immediate effects) and after 15 and 30-day period of daily application (long term effects). Results obtained were statistically analyzed using the ANOVA parametric test. In the rheological study, the formulations number 1, 2, and 5 were considered more stable than the others. Moreover, the formulations number 5 and 6 showed the best sensorial attributes, and for this reason the formulation number 5 was selected for the efficacy study. The evaluation of immediate effects of formulations showed that the application of the formulations studied in the volar forearm increased the water content of the stratum corneum, while only the formulations containing the peptides provoked an increased in this content, when face skin was analyzed. When the volar forearm skin microrelief was analyzed, the application of all the formulations studied provoked a significant decrease in skin roughness (Ser) and depth of roughness (Rt), furthermore only the formulation supplemented with the DTP showed a significant increase in the skin smoothness (Sesm). In the evaluation of long term effects, all formulations provoked a significant reduction in TEWL values and the formulations containing the peptides studied provoked a significant increase in the water content of the stratum corneum of eye corner; when the volar forearms and the cheek region were analyzed, all the formulations containing or not the peptides studied, provoked an increase in this parameter. When the skin microrelief was analyzed, only the formulation supplemented with AHP or ATP or DTP, provoked a significant increase in the skin smoothness (Sesm), when applied in the cheek region. The application of formulations containing the peptides studied in the volar forearms provoked a significant decrease in the skin anisotropy. The formulation nº 5 (F5) containing the peptides studied, showed immediate effects on skin hydration, roughness and smoothness, and also long-term effects on skin hydration, smoothness and mechanical properties, which demonstrated the importance of the use of these peptides in cosmetic formulations with hydration and anti-aging purposes.

Yaku'amide A shows a unique inhibitory profile against a series of 39 human cancer cell lines (JFCR39). In our efforts to synthesize yaku'amide A, we have optimized our regioselective base-free aminohydroxylation method with a series of nitrogen sources, developed a chiral reagent-mediated aminohydroxylation strategy and chemoselective deprotections of the resulting aminohydroxylation product, and explored a stereospecific E2 dehydration and O-N acyl transfer sequence. In addition, we have prepared the right-hand tetrapeptide and the NTA subunit. For our bulky α,β-dehydroamino acids project, we have developed strategies to incorporate α,β-dehydroamino acids such as ΔVal and ΔEnv into small synthetic peptides via Solid Phase Peptide Synthesis (SPPS). We have also prepared two analogues of a monomeric helical peptide with 13 residues.

Structural studies of peptides has relevance for various applications, like, in de novo design of proteins, in designing better catalysts for organic synthesis, in structure based drug design, in the design and construction of synthetic protein mimics and in building novel materials via supramolecular self assembly. Crystal structure determination of peptides is expected to provide information about their static structure, mode of aggregation, solvation and hydrogen bond interactions of the sequences in the solid state. Comparison and analysis of the related structures from the database analysis could provide information about sequence dependent conformational features, which eventually would act as precursor for de novo protein design. Self assembling processes are common throughout nature and technology. Living cells self assemble, and understanding life will therefore require an understanding of self assembly. Supramolecular chemistry has become an area of intense research, partly inspired by biological ensembles in nature, such as collagen and enzymes or protein assemblies in general. Understanding, inducing, and directing such self assembling processes are key to unraveling the progressive emergence of complex matter. Most of the drugs available today have a broad spectrum of action in that they can act on more than one receptor and the mechanism of action of these drugs are poorly understood. Homology modeling of receptors and docking studies with drug molecules (both peptides and non-peptides) would result in a better understanding of the mechanism of drug-receptor binding thus resulting in the design of more specific and effective drugs. This thesis reports the results of X-ray crystallographic studies of ten molecules listed below (Ter: terephthalic acid) and the molecular model of cholecystokinin type 1 receptor (CCK1R). The abbreviations used for the sequences are given in parenthesis. Boc-Gly-Dpg-Gly-Leu-OMe (GDGL), C24H44N4O7 Boc-Val-Ala-Leu-Dpg-Val-Ala-Leu-Val-Ala-Leu-Dpg-Val-Ala-Leu-OMe (VAL14), C78H14 2N14O17 MeO-Leu-Ter-Leu-OMe (LTeL), C22H32N2O6 MeO-DLeu-Ter-DLeu-OMe (DLTeDL), C22H32N2O6 MeO-Ile-Ter-Ile-OMe (ITeI), C22H32N2O6 MeO-Aib-Ter-Aib-OMe (UTeU), C18H24N2O6 Tyr-Aib-Tyr-Val (YUYV), C27H36N4O7 Tyr-Aib-Ala (YUA), C16H23N3O5 Z-Gly-Gly-Val (ZGGV), C17 H23 N3 O6 DL-4-benzamido-N, N-dipropylglutaramic acid (proglumide), C18 H26 N2 O4 Results from the Dpg containing peptide sequences helped to further the understanding of conformational preferences of this residue. The crystallographic studies on the peptide sequence, which forms a supramolecular triple helix and four pseudopeptide sequences, which adopt supramolecular ladder conformations has provided substantial information on the role of non covalent interactions in supramolecular self assembly. Crystal structure of a Gly-Gly containing tripeptide and database analysis has provided insights into the conformations adopted by this segment in peptides and proteins. The docking of the crystal structure of proglumide, an antagonist of CCK1R has led to the understanding of the mechanism of its interaction with CCK1R.

O objetivo desta pesquisa foi o desenvolvimento, avaliação da estabilidade e da eficácia de formulações cosméticas contendo acetil hexapeptídeo-3 (AHP) ou acetil tetrapeptídeo-5 (ATP) ou mistura de di e tripeptídeos (DTP). Para tal, foram desenvolvidas 6 formulações à base hidroxietilcelulose (HEC), metilfenil polissiloxano (MFP) e álcool batílico e lecitina de soja (ABL) (F1), ou de HEC, MFP, ésteres de polietilenoglicol com cadeia alquílica com 12 a 20 carbonos (EP) (F2), ou de polímero de acrilatos (PA), MFP e ABL (F3) ou de PA, MFP e EP (F4), ou de HEC e MFP (F5) ou de PA e MFP (F6), contendo ou não (veículo) 10% de AHP, ou 10% de ATP ou 4% de DTP, as quais foram submetidas a um estudo de estabilidade física por determinação da reologia e a uma avaliação sensorial. Para o estudo de eficácia dos peptídeos objeto de estudo as formulações foram aplicadas nos antebraços, na face, na região dos olhos e da boca de 60 voluntárias, sendo realizadas medidas do conteúdo aquoso do estrato córneo, da perda de água transepidérmica (TEWL), do micro-relevo cutâneo e das propriedades mecânicas da pele (anisotropia), antes (basal), após 2 horas da aplicação única (efeitos imediatos) e após 15 e 30 dias de uso contínuo (efeitos a longo prazo) das formulações. Os dados foram analisados estatisticamente pelo teste paramétrico análise de variância. Nos testes de estabilidade física, as formulações de nos 1, 2 e 5 foram consideradas as mais estáveis e as formulações de nº 5 e 6 foram as que apresentaram melhor sensorial, sendo, portanto, selecionada a formulação de nº 5 como veículo para os estudos de eficácia. Na avaliação dos efeitos imediatos na pele, observou-se que as formulações estudadas proporcionaram um aumento significativo no conteúdo aquoso do estrato córneo dos antebraços, enquanto que, somente as formulações acrescidas dos peptídeos provocaram um aumento significativo neste conteúdo na face. Em relação ao micro-relevo cutâneo, as formulações contendo ou não os peptídeos provocaram uma redução do parâmetro Ser (aspereza da pele) e Rt (rugosidade) nos antebraços e a formulação contendo DTP provocou um efeito imediato significativo no parâmetro Sesm (textura da pele). Na avaliação dos efeitos a longo prazo, as formulações ocasionaram uma diminuição significativa na TEWL e as acrescidas dos peptídeos objetos de estudo proporcionaram um aumento significativo no conteúdo aquoso do estrato córneo na região dos olhos, enquanto que nos antebraços e na região da boca todas as formulações acrescidas ou não dos peptídeos ocasionaram um aumento nesse parâmetro. Em relação ao micro-relevo cutâneo, a aplicação das formulações contendo os peptídeos, na região da boca, provocou um aumento significativo nos valores de Sesm. A aplicação das formulações contendo os peptídeos objeto de estudo, nos antebraços ocasionou uma diminuição significativa da anisotropia da pele. A formulação nº 5 contendo os peptídeos objeto de estudo, apresentou efeitos imediatos na hidratação, aspereza, rugosidade e textura da pele e, efeitos a longo prazo, na hidratação, textura e anisotropia da pele, mostrando a importância do emprego destes peptídeos em produtos cosméticos com finalidades hidratantes e antienvelhecimento. / The aim of this study was the development, evaluation of the stability and the efficacy of cosmetic formulations containing acetyl hexapeptide-3 (AHP), or acetyl tetrapeptide-5 (ATP) or mixture of di- and peptides (DTP). For this purpose the 6 following formulations (F) were developed: F1- containing hydroxyethyl cellulose (HEC), methylphenyl polysiloxane (MPPS), batyl alcohol, and lecithin (BAL); F2- HEC, MPPS and C12-C20 PEG-8 alkyl ester (AE); F3- acrylate copolymer (AC), MPPS and BAL; F4- AC, MPPS and AE; F5- HEC and MPPS and F6- AC and MPPS, which were supplemented or not (vehicle) with 10% of AHP or 10% of ATP or 4% of DTP and submitted to the evaluation of physical stability by the determination of the rheologic behavior and also to the sensorial analysis. For clinical efficacy studies, formulations were applied to the volar forearm and to the face skin (eye corner and cheek region), of 60 female volunteers. Skin conditions in terms of skin moisture, transepidermal water loss (TEWL), skin mechanical properties (anisotropy) and skin microrelief, before and after 2 hours (immediate effects) and after 15 and 30-day period of daily application (long term effects). Results obtained were statistically analyzed using the ANOVA parametric test. In the rheological study, the formulations number 1, 2, and 5 were considered more stable than the others. Moreover, the formulations number 5 and 6 showed the best sensorial attributes, and for this reason the formulation number 5 was selected for the efficacy study. The evaluation of immediate effects of formulations showed that the application of the formulations studied in the volar forearm increased the water content of the stratum corneum, while only the formulations containing the peptides provoked an increased in this content, when face skin was analyzed. When the volar forearm skin microrelief was analyzed, the application of all the formulations studied provoked a significant decrease in skin roughness (Ser) and depth of roughness (Rt), furthermore only the formulation supplemented with the DTP showed a significant increase in the skin smoothness (Sesm). In the evaluation of long term effects, all formulations provoked a significant reduction in TEWL values and the formulations containing the peptides studied provoked a significant increase in the water content of the stratum corneum of eye corner; when the volar forearms and the cheek region were analyzed, all the formulations containing or not the peptides studied, provoked an increase in this parameter. When the skin microrelief was analyzed, only the formulation supplemented with AHP or ATP or DTP, provoked a significant increase in the skin smoothness (Sesm), when applied in the cheek region. The application of formulations containing the peptides studied in the volar forearms provoked a significant decrease in the skin anisotropy. The formulation nº 5 (F5) containing the peptides studied, showed immediate effects on skin hydration, roughness and smoothness, and also long-term effects on skin hydration, smoothness and mechanical properties, which demonstrated the importance of the use of these peptides in cosmetic formulations with hydration and anti-aging purposes.