The development of chiral organometallics for asymmetric synthesis is a topic of significant research in the recent past. The most studied in this class are the chiral organolithium reagents with many reported examples. The primary focus of our research is the development of C_α-chiral Grignard reagents, where the metal bearing α-carbon is the sole source of chirality. Examples of such Grignard reagents are rare owing to the problems associated with their synthesis, and their low configurational stability. We have studied these problems in three different modules of this project. Reactions of 1-magnesio-2,2-diphenyl-cyclopropylcarbonitrile with carbon electrophiles are first attempted in order to expand the utility of this configurationally stable C_α-chiral Grignard reagent in asymmetric synthesis. This reagent has been shown to be non-reactive towards carbon electrophiles at low temperatures. Consequently, we attempt to enhance the reactivity of this compound through two different approaches, Lewis-base activation and the "ate-complex" generation. The Magnesium/Halogen (Mg/X) exchange reactions have been shown to be extremely useful in the synthesis of complex Aryl, alkenyl (sp²) and alkynyl (sp) Grignard reagents. Examples of Mg/X exchange reactions of Alkyl (sp³) halides are, however, rare. Even more rare are such examples with secondary and tertiary alkyl halides, justifying the relative paucity of chiral Grignard reagents. In this module of our project, we study the Mg/X exchange reactions on secondary alkyl halides possessing a γ-hydroxyl group, as an internal activator for such Mg/X exchange reactions. Enantiomerization pathways of chiral organolithium compounds have been widely studied. However, few such studies have been performed on chiral Grignard reagents. In this module of the project, we studied the solvent assisted enantiomerization mechanism of the C_α-chiral 1-magnesio-2,2-diphenyl-cyclopropylcarbonitrile. Rate constant for the enantiomerization of this compound was measured in three different ethereal solvents to study the effect of solvent on the configurational stability. Finally, the order of the enantiomerization process with respect to [Et₂O] was studied in order to predict the mechanism of this process in Et₂O solvent. Our kinetic studies on the enantiomerization process provided us with a definitive picture for the enantiomerization of the C_α-chiral 1-magnesio-2,2-diphenyl-cyclopropylcarbonitrile, where solvation of the Grignard reagent preceded an ion-pair separation step which eventually lead to enantiomerization of the Grignard species. However, the precise structure of all the involved solvated intermediates could not be determined as kinetics was not able to distinguish between these intermediates. We next performed computational calculations to study the effect of solvation on the analogous 1-magnesio-cyclopropylcarbonitrile in order to address the unanswered questions from our kinetic studies. / Ph. D.

É sabido que: (i) as sínteses individual e múltipla (manual e automática), bem como a construção de bibliotecas e micro-arranjos de peptídeos sintéticos empregam a metodologia da fase sólida (SPFS); (ii) apesar do desenvolvimento atual desta metodologia sintética, os químicos de peptídeos continuam se deparando com problemas e limitações inerentes a ela; (iii) muitos trabalhos relatam a sua agilização pelo uso de altas temperaturas, mas poucos revelam preocupação com a integridade quiral dos peptídeos-alvo. Portanto, o presente trabalho objetivou dar continuidade à nossa investigação pioneira das: 1) incidência da enantiomerização dos aminoácidos e/ou de outras reações secundárias nas SPFS passo a passo de peptídeos a 60 °C; 2) viabilidade de realização de todas as etapas da síntese convergente em fase sólida (SCPFS) a 60 °C. Em relação ao tópico 1, os peptídeos-alvo escolhidos tinham tamanho e sequência variáveis que incluiam os aminoácidos trifuncionais problemáticos Cys, Ser, His, Met e Trp. Todos eles foram obtidos por SPFS passo a passo convencional e a 60 °C usando aquecimento convencional e micro-ondas. A identificação e a quantificação dos isômeros contaminantes foram feitas com a ajuda de padrões resultantes da SPFS passo a passo convencional e de métodos analíticos (RP-HPLC, LC-ESI/MS, CE e análise quiral de aminoácidos) em condições estabelecidas por nós. Foi constatado que: (i) as nossas condições de acoplamento são mais econômicas que as usuais, pois empregam menor concentração e excesso molar de N-acil-aminoácidos; (ii) nelas, a SPFS a 60 °C usando o aquecimento convencional é simples, prática, de custo relativamente baixo, demanda ½ do tempo da SPFS convenciona e não compromete significativamente a integridade quiral dos aminoácidos; (iii) nas condições similares, a SPFS passo a passo a 60 °C assistida por micro-ondas é mais rápida (realizada em ¼ do tempo gasto na SPFS convencional), porém mais cara e acompanhada de aumento significativo da enantiomerização da Cys; (iv) a mistura 25% DMSO/tolueno, nunca antes utilizada na SPFS assistida por micro-ondas, favoreceu a formação de contaminantes contendo Met oxidadas a sulfóxidos durante as sínteses do fragmento CCK24-33NS, mas o mesmo ocorreu quando DMF foi usado nas sínteses da CCK-33 NS; (v) outras reações secundárias típicas da SPFS passo a passo não foram intensificadas significativamente nas nossas condições sintéticas a 60 °C. Quanto ao tópico 2, foi escolhida a CCK-33 NS como modelo peptídico. Foi constatado que: (i) a etapa de obtenção dos fragmentos peptídicos protegidos que atuariam como doadores de acila (D.A.) e aceptor de acila (A.A.) de partida pode ser ágil e bem sucedida pela SPFS passo a passo a 60 °C nas nossas condições experimentais usando o aquecimento convencional; (ii) DMF, NMP e 25% DMSO/Tolueno foram adequados à solubilização dos fragmentos D.A. e dos reagentes necessários à sua ativação a 60 °C; (iii) a 60 °C, tais solventes também intumesceram satisfatoriamente a CCK24-33NS-resina Rink amida, A.A. de partica; (iv) o aquecimento convencional permitiu que algumas reações de acoplamento entre os D.A. e A.A. escolhidos fossem realizadas com sucesso; na maioria dos casos em que isso não ocorreu, o uso combinado das micro-ondas e agitação sob atmosfera inerte mediaram a formação do produto desejado; (v) a natureza dos fragmentos D.A. e A.A. é fator limitante na SCPFS, mesmo a 60 °C e usando o aquecimento convencional ou as micro-ondas, e, portanto, ele precisa ser melhor estudado. / It is well known that: (i) individual and multiple peptide syntheses (manual and automatic) as well as construction of synthetic peptide libraries and micro-arrays are all based on solid phase chemistry (SPPS); (ii) despite the current development of such synthetic methodology, peptide chemists are still facing its problems and inherent limitations; (iii) many previous works employed high temperatures to accelerate stepwise SPPS, but only a few showed concern about the preservation of the chiral integrity of the target peptide. Therefore, the main goal of the present work was to continue our pioneering investigation of: 1) the incidence of amino acids enantiomerization and/or other side-reactions in the stepwise SPPS at 60°C, 2) the viability of performing all steps of the convergent synthesis on a solid support (CSPPS) at 60°C. With regard to the topic 1, the peptides chosen as targets had variable size and sequence, which included the tricky trifunctional amino acids Cys, Ser, His, Met and Trp. The peptides were synthesized by conventional stepwise SPFS and at 60 °C using conventional or microwave heating. Identification and quantification of the contaminant isomers was done with the aid of standards resultant from conventional stepwise SPPS and of analytical methods (RP-HPLC, LC-ESI/MS, CE and chiral amino acids analysis) in conditions established in our laboratory. It was shown that: (i) our coupling conditions are cheaper than the usual ones as they employ lower concentration and excess of N-acyl-amino acids; (ii) under them, stepwise SPPS at 60 °C using the conventional heating is simple, practical, relatively low-cost, demands half of the time required by conventional stepwise SPPS and does not cause the enhancement of amino acids enantiomerization; (iii) under similar conditions, microwave-assisted stepwise SPPS at 60 °C is faster (it demands only one-fourth of the time spent in the conventional stepwise SPPS), but it is more expensive and causes significant damage specially to the chiral integrity of Cys; (iv) the binary mixture 25% DMSO/toluene, never used earlier in microwave-assisted stepwise SPPS, led to the formation of contaminants with Met oxidized to its sulfoxides during the synthesis of CCK24-33NS; however, it also occurred when DMF was used in the synthesis of CCK-33 NS; (v) other side reactions typical of stepwise SPPS were not significantly intensified under our conditions at 60 °C. Concerning to the topic 2, CCK-33 NS was chosen as the model peptide. It was shown that: (i) the synthesis of the protected peptides that would act as acyl donor (A.D.) or as the starting acyl aceptor (A.A.) can be fast and successfully achieved at 60 °C under our experimental conditions using conventional heating; (ii) DMF, NMP and 25% DMSO/toluene dissolved all A.D. and the reagents required for their activation at 60 °C; (iii) at this temperature, such solvents were also able to properly swell CCK24-33NS-Rink amide resin, the starting A.A.; (iv) the conventional heating allowed for some coupling reactions between A.D. and A.A., but in most cases in which it did not occur, the combined use of microwaves and stirring under inert atmosphere mediated the formation of the desired products; (v) the nature of fragments A.D. and A.A. is a limiting factor in the CSPPS even at 60 °C and using the conventional or microwave heating; therefore, it should be further studied.

É sabido que: (i) as sínteses individual e múltipla (manual e automática), bem como a construção de bibliotecas e micro-arranjos de peptídeos sintéticos empregam a metodologia da fase sólida (SPFS); (ii) apesar do desenvolvimento atual desta metodologia sintética, os químicos de peptídeos continuam se deparando com problemas e limitações inerentes a ela; (iii) muitos trabalhos relatam a sua agilização pelo uso de altas temperaturas, mas poucos revelam preocupação com a integridade quiral dos peptídeos-alvo. Portanto, o presente trabalho objetivou dar continuidade à nossa investigação pioneira das: 1) incidência da enantiomerização dos aminoácidos e/ou de outras reações secundárias nas SPFS passo a passo de peptídeos a 60 °C; 2) viabilidade de realização de todas as etapas da síntese convergente em fase sólida (SCPFS) a 60 °C. Em relação ao tópico 1, os peptídeos-alvo escolhidos tinham tamanho e sequência variáveis que incluiam os aminoácidos trifuncionais problemáticos Cys, Ser, His, Met e Trp. Todos eles foram obtidos por SPFS passo a passo convencional e a 60 °C usando aquecimento convencional e micro-ondas. A identificação e a quantificação dos isômeros contaminantes foram feitas com a ajuda de padrões resultantes da SPFS passo a passo convencional e de métodos analíticos (RP-HPLC, LC-ESI/MS, CE e análise quiral de aminoácidos) em condições estabelecidas por nós. Foi constatado que: (i) as nossas condições de acoplamento são mais econômicas que as usuais, pois empregam menor concentração e excesso molar de N-acil-aminoácidos; (ii) nelas, a SPFS a 60 °C usando o aquecimento convencional é simples, prática, de custo relativamente baixo, demanda ½ do tempo da SPFS convenciona e não compromete significativamente a integridade quiral dos aminoácidos; (iii) nas condições similares, a SPFS passo a passo a 60 °C assistida por micro-ondas é mais rápida (realizada em ¼ do tempo gasto na SPFS convencional), porém mais cara e acompanhada de aumento significativo da enantiomerização da Cys; (iv) a mistura 25% DMSO/tolueno, nunca antes utilizada na SPFS assistida por micro-ondas, favoreceu a formação de contaminantes contendo Met oxidadas a sulfóxidos durante as sínteses do fragmento CCK24-33NS, mas o mesmo ocorreu quando DMF foi usado nas sínteses da CCK-33 NS; (v) outras reações secundárias típicas da SPFS passo a passo não foram intensificadas significativamente nas nossas condições sintéticas a 60 °C. Quanto ao tópico 2, foi escolhida a CCK-33 NS como modelo peptídico. Foi constatado que: (i) a etapa de obtenção dos fragmentos peptídicos protegidos que atuariam como doadores de acila (D.A.) e aceptor de acila (A.A.) de partida pode ser ágil e bem sucedida pela SPFS passo a passo a 60 °C nas nossas condições experimentais usando o aquecimento convencional; (ii) DMF, NMP e 25% DMSO/Tolueno foram adequados à solubilização dos fragmentos D.A. e dos reagentes necessários à sua ativação a 60 °C; (iii) a 60 °C, tais solventes também intumesceram satisfatoriamente a CCK24-33NS-resina Rink amida, A.A. de partica; (iv) o aquecimento convencional permitiu que algumas reações de acoplamento entre os D.A. e A.A. escolhidos fossem realizadas com sucesso; na maioria dos casos em que isso não ocorreu, o uso combinado das micro-ondas e agitação sob atmosfera inerte mediaram a formação do produto desejado; (v) a natureza dos fragmentos D.A. e A.A. é fator limitante na SCPFS, mesmo a 60 °C e usando o aquecimento convencional ou as micro-ondas, e, portanto, ele precisa ser melhor estudado. / It is well known that: (i) individual and multiple peptide syntheses (manual and automatic) as well as construction of synthetic peptide libraries and micro-arrays are all based on solid phase chemistry (SPPS); (ii) despite the current development of such synthetic methodology, peptide chemists are still facing its problems and inherent limitations; (iii) many previous works employed high temperatures to accelerate stepwise SPPS, but only a few showed concern about the preservation of the chiral integrity of the target peptide. Therefore, the main goal of the present work was to continue our pioneering investigation of: 1) the incidence of amino acids enantiomerization and/or other side-reactions in the stepwise SPPS at 60°C, 2) the viability of performing all steps of the convergent synthesis on a solid support (CSPPS) at 60°C. With regard to the topic 1, the peptides chosen as targets had variable size and sequence, which included the tricky trifunctional amino acids Cys, Ser, His, Met and Trp. The peptides were synthesized by conventional stepwise SPFS and at 60 °C using conventional or microwave heating. Identification and quantification of the contaminant isomers was done with the aid of standards resultant from conventional stepwise SPPS and of analytical methods (RP-HPLC, LC-ESI/MS, CE and chiral amino acids analysis) in conditions established in our laboratory. It was shown that: (i) our coupling conditions are cheaper than the usual ones as they employ lower concentration and excess of N-acyl-amino acids; (ii) under them, stepwise SPPS at 60 °C using the conventional heating is simple, practical, relatively low-cost, demands half of the time required by conventional stepwise SPPS and does not cause the enhancement of amino acids enantiomerization; (iii) under similar conditions, microwave-assisted stepwise SPPS at 60 °C is faster (it demands only one-fourth of the time spent in the conventional stepwise SPPS), but it is more expensive and causes significant damage specially to the chiral integrity of Cys; (iv) the binary mixture 25% DMSO/toluene, never used earlier in microwave-assisted stepwise SPPS, led to the formation of contaminants with Met oxidized to its sulfoxides during the synthesis of CCK24-33NS; however, it also occurred when DMF was used in the synthesis of CCK-33 NS; (v) other side reactions typical of stepwise SPPS were not significantly intensified under our conditions at 60 °C. Concerning to the topic 2, CCK-33 NS was chosen as the model peptide. It was shown that: (i) the synthesis of the protected peptides that would act as acyl donor (A.D.) or as the starting acyl aceptor (A.A.) can be fast and successfully achieved at 60 °C under our experimental conditions using conventional heating; (ii) DMF, NMP and 25% DMSO/toluene dissolved all A.D. and the reagents required for their activation at 60 °C; (iii) at this temperature, such solvents were also able to properly swell CCK24-33NS-Rink amide resin, the starting A.A.; (iv) the conventional heating allowed for some coupling reactions between A.D. and A.A., but in most cases in which it did not occur, the combined use of microwaves and stirring under inert atmosphere mediated the formation of the desired products; (v) the nature of fragments A.D. and A.A. is a limiting factor in the CSPPS even at 60 °C and using the conventional or microwave heating; therefore, it should be further studied.