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Jarbas Magalhães Resendehttp://lattes.cnpq.br/4995867883627482Jarbas Magalhães ResendeAdolfo Henrique de Moraes SilvaTiago Antônio da Silva BrandãoFábio Ceneviva Lacerda de AlmeidaCláudio Francisco Tormenahttp://lattes.cnpq.br/6047367281104531Lucas Raposo Carvalho2023-03-30T17:20:22Z2023-03-30T17:20:22Z2022-10-26http://hdl.handle.net/1843/51378https://orcid.org/0000-0002-6476-4531Antimicrobial peptides (AMPs) are promising drug candidates when fighting bacterial resistance, being molecules with many mechanisms of action. Furthermore, the skin secretion of anurans and spider poisons are rich sources of AMPs, and phylloseptine PS-O1, alongside LyeTx I-b, are peptides of interest to our research group. Peptide alteration strategies, such as post-translational modifications and residue insertion, aim to improve the biological properties of these drug candidates. As such, in this work, six compounds were synthesized, being two peptides (PS-O1 and R1G2-PS-O1) and two propargylic derivatives ([Pra1 ]PS-O1 and R[Pra]PS-O1) by Fmoc-SPPS and two glucotriazole peptides (PS-O1 GtP and R1A2-PS-O1 GtP) using the CuAAC reaction. Two peptides, LyeTx I-bcys and LyeTx I-bPEG, were synthesized by collaborator Júlio César Moreira Brito. PS-O1 derivatives were synthesized (8.7 to 26.5% yield), purified by RP-HPLC (73.4 to 91.5% yield), with purities ranging from 92% to 98%, and analyzed by MALDI-TOF-MS. Circular dichroism was used to analyze their conformational prefer ences in the presence of TFE/H2O mixtures, SDS and DPC micelles, and POPC and POPC/POPG 3:1 vesicles, indicating α-helical structural motifs starting from specific concentrations for all media except POPC LUVs. The three-dimensional structure of PS-O1, PS-O1 GtP, R1G2-PS-O1, R1A2-PS-O1 GtP, LyeTx I-bcys, and LyeTx I-bPEG was determined by 2D NMR experiments, revealing various α-helical percentages and positions. While the introduction of a glucose unit increased structure formation for PS-O1, an opposite effect was observed for R1G2-PS-O1, being potentially attributed to different helix stabilizing interactions at the N -terminus. A high degree of structural similarity between LyeTx I-bcys and LyeTx I-bPEG was observed. Finally, antifungal biological studies for PS-O1 derivatives show that R1G2-PS-O1 is considerably more active than the other three, supporting structural data obtained by NMR analyses.Peptídeos antimicrobianos (PAMs) são promissores candidatos a fármacos para combater a resistência bacteriana, sendo moléculas com mecanismos de ação diversos. Além disso, a secreção cutânea de anuros e venenos de aranhas são fontes abundantes de PAMs e a filloseptina PS-O1, assim como a LyeTx I-b, são peptídeos de interesse do nosso grupo de pesquisa. Estratégias de alterações de peptídeos, como modificações pós-translacionais e inserção de resíduos visam melhorar propriedades farmacológicas desses potenciais fármacos. Sendo assim, neste trabalho, foram sintetizados seis compostos, dois peptídeos (PS-O1 e R1G2-PS-O1) e dois derivados propargílicos ([Pra1]PS-O1 e R[Pra]PS O1) por Fmoc-SPFS e dois glicotriazol-peptídeos (PS-O1 GtP e R1A2-PS-O1 GtP) por CuAAC. Os derivados LyeTx I-bcys e LyeTx I-bPEG foram sintetizados pelo colaborador Júlio César Moreira Brito. A síntese dos derivados da PS-O1 tiveram rendimentos de síntese entre 8,7 e 26,5% e, de purificação, entre 73,4 e 91,5%, com graus de pureza entre 91 e 98%, verificados por CLAE-FR. Todos os derivados sintetizados foram caracterizados por MALDI-TOF-MS e tiveram suas preferências conformacionais analisadas por dicroísmo circular na presença de misturas TFE:H2O, micelas de SDS e DPC e vesículas de POPC e POPC:POPG 3:1, indicando a formação de estruturas α-helicoidais a partir de determinados valores de concentração para todos os meios exceto LUVs de POPC. A estrutura tridimensional dos peptídeos PS-O1, PS-O1 GtP, R1G2-PS-O1, R1A2-PS-O1 GtP, LyeTx I-bcys e LyeTx I-bPEG foram determinadas por experimentos de RMN, revelando diferentes porcentages e localizações de estruturas α-helicoidais. Observou-se que, enquanto a introdução da unidade glicosídica aumentou a estruturação para a PS-O1, um efeito contrário foi observado para a R1G2-PS-O1, sendo potencialmente atribuído à diferentes interações estabilizantes de hélice próximas à porção N -terminal. Um alto grau de similaridade estrutural entre os peptídeos LyeTx I-bcys e LyeTx I-bPEG foi observado. Por fim, estudos de atividade anti-fúngica para os derivados da PS-O1 mostram que a R1G2-PS-O1 apresentou atividade pronunciadamente superior aos outros, corroborando dados estruturais observados por RMN.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas GeraisCAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorFINEP - Financiadora de Estudos e Projetos, Financiadora de Estudos e ProjetosOutra AgênciaengUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em QuímicaUFMGBrasilICX - DEPARTAMENTO DE QUÍMICAhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessQuímica orgânicaPeptídios – SínteseProdutos de ação antimicrobianaRessonância magnética nuclearArgininaFormação de anéis (Química)Cromatografia liquida de alta eficiênciaTestes biológicosAtividade antifúngicaCitotoxidade de mediação celularAgentes antibacterianosAntimicrobial peptidesPhylloseptinsLyeTx I-bNuclear magnetic resonance - NMR spectroscopyPeptide-membrane interactionsGlucotriazole-peptidesPEGylated peptidesArginineStructural, membrane interaction and biological studies of PS-O1 and LyeTx I-b peptide derivativesEstudos estruturais, de interação com membranas e biológicos de derivados peptídicos da PS-O1 e LyeTx I-binfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGLICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/51378/5/license.txtcda590c95a0b51b4d15f60c9642ca272MD55CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufmg.br/bitstream/1843/51378/2/license_rdfcfd6801dba008cb6adbd9838b81582abMD52ORIGINALLucas_Raposo_PhD_Thesis.pdfLucas_Raposo_PhD_Thesis.pdfArquivo da tese, em formato pdfapplication/pdf12855457https://repositorio.ufmg.br/bitstream/1843/51378/4/Lucas_Raposo_PhD_Thesis.pdfd11f49d273e475786dbf7daf81b1f51bMD541843/513782023-03-30 14:20:23.105oai:repositorio.ufmg.br:1843/51378TElDRU7Dh0EgREUgRElTVFJJQlVJw4fDg08gTsODTy1FWENMVVNJVkEgRE8gUkVQT1NJVMOTUklPIElOU1RJVFVDSU9OQUwgREEgVUZNRwoKQ29tIGEgYXByZXNlbnRhw6fDo28gZGVzdGEgbGljZW7Dp2EsIHZvY8OqIChvIGF1dG9yIChlcykgb3UgbyB0aXR1bGFyIGRvcyBkaXJlaXRvcyBkZSBhdXRvcikgY29uY2VkZSBhbyBSZXBvc2l0w7NyaW8gSW5zdGl0dWNpb25hbCBkYSBVRk1HIChSSS1VRk1HKSBvIGRpcmVpdG8gbsOjbyBleGNsdXNpdm8gZSBpcnJldm9nw6F2ZWwgZGUgcmVwcm9kdXppciBlL291IGRpc3RyaWJ1aXIgYSBzdWEgcHVibGljYcOnw6NvIChpbmNsdWluZG8gbyByZXN1bW8pIHBvciB0b2RvIG8gbXVuZG8gbm8gZm9ybWF0byBpbXByZXNzbyBlIGVsZXRyw7RuaWNvIGUgZW0gcXVhbHF1ZXIgbWVpbywgaW5jbHVpbmRvIG9zIGZvcm1hdG9zIMOhdWRpbyBvdSB2w61kZW8uCgpWb2PDqiBkZWNsYXJhIHF1ZSBjb25oZWNlIGEgcG9sw610aWNhIGRlIGNvcHlyaWdodCBkYSBlZGl0b3JhIGRvIHNldSBkb2N1bWVudG8gZSBxdWUgY29uaGVjZSBlIGFjZWl0YSBhcyBEaXJldHJpemVzIGRvIFJJLVVGTUcuCgpWb2PDqiBjb25jb3JkYSBxdWUgbyBSZXBvc2l0w7NyaW8gSW5zdGl0dWNpb25hbCBkYSBVRk1HIHBvZGUsIHNlbSBhbHRlcmFyIG8gY29udGXDumRvLCB0cmFuc3BvciBhIHN1YSBwdWJsaWNhw6fDo28gcGFyYSBxdWFscXVlciBtZWlvIG91IGZvcm1hdG8gcGFyYSBmaW5zIGRlIHByZXNlcnZhw6fDo28uCgpWb2PDqiB0YW1iw6ltIGNvbmNvcmRhIHF1ZSBvIFJlcG9zaXTDs3JpbyBJbnN0aXR1Y2lvbmFsIGRhIFVGTUcgcG9kZSBtYW50ZXIgbWFpcyBkZSB1bWEgY8OzcGlhIGRlIHN1YSBwdWJsaWNhw6fDo28gcGFyYSBmaW5zIGRlIHNlZ3VyYW7Dp2EsIGJhY2stdXAgZSBwcmVzZXJ2YcOnw6NvLgoKVm9jw6ogZGVjbGFyYSBxdWUgYSBzdWEgcHVibGljYcOnw6NvIMOpIG9yaWdpbmFsIGUgcXVlIHZvY8OqIHRlbSBvIHBvZGVyIGRlIGNvbmNlZGVyIG9zIGRpcmVpdG9zIGNvbnRpZG9zIG5lc3RhIGxpY2Vuw6dhLiBWb2PDqiB0YW1iw6ltIGRlY2xhcmEgcXVlIG8gZGVww7NzaXRvIGRlIHN1YSBwdWJsaWNhw6fDo28gbsOjbywgcXVlIHNlamEgZGUgc2V1IGNvbmhlY2ltZW50bywgaW5mcmluZ2UgZGlyZWl0b3MgYXV0b3JhaXMgZGUgbmluZ3XDqW0uCgpDYXNvIGEgc3VhIHB1YmxpY2HDp8OjbyBjb250ZW5oYSBtYXRlcmlhbCBxdWUgdm9jw6ogbsOjbyBwb3NzdWkgYSB0aXR1bGFyaWRhZGUgZG9zIGRpcmVpdG9zIGF1dG9yYWlzLCB2b2PDqiBkZWNsYXJhIHF1ZSBvYnRldmUgYSBwZXJtaXNzw6NvIGlycmVzdHJpdGEgZG8gZGV0ZW50b3IgZG9zIGRpcmVpdG9zIGF1dG9yYWlzIHBhcmEgY29uY2VkZXIgYW8gUmVwb3NpdMOzcmlvIEluc3RpdHVjaW9uYWwgZGEgVUZNRyBvcyBkaXJlaXRvcyBhcHJlc2VudGFkb3MgbmVzdGEgbGljZW7Dp2EsIGUgcXVlIGVzc2UgbWF0ZXJpYWwgZGUgcHJvcHJpZWRhZGUgZGUgdGVyY2Vpcm9zIGVzdMOhIGNsYXJhbWVudGUgaWRlbnRpZmljYWRvIGUgcmVjb25oZWNpZG8gbm8gdGV4dG8gb3Ugbm8gY29udGXDumRvIGRhIHB1YmxpY2HDp8OjbyBvcmEgZGVwb3NpdGFkYS4KCkNBU08gQSBQVUJMSUNBw4fDg08gT1JBIERFUE9TSVRBREEgVEVOSEEgU0lETyBSRVNVTFRBRE8gREUgVU0gUEFUUk9Dw41OSU8gT1UgQVBPSU8gREUgVU1BIEFHw4pOQ0lBIERFIEZPTUVOVE8gT1UgT1VUUk8gT1JHQU5JU01PLCBWT0PDiiBERUNMQVJBIFFVRSBSRVNQRUlUT1UgVE9ET1MgRSBRVUFJU1FVRVIgRElSRUlUT1MgREUgUkVWSVPDg08gQ09NTyBUQU1Cw4lNIEFTIERFTUFJUyBPQlJJR0HDh8OVRVMgRVhJR0lEQVMgUE9SIENPTlRSQVRPIE9VIEFDT1JETy4KCk8gUmVwb3NpdMOzcmlvIEluc3RpdHVjaW9uYWwgZGEgVUZNRyBzZSBjb21wcm9tZXRlIGEgaWRlbnRpZmljYXIgY2xhcmFtZW50ZSBvIHNldSBub21lKHMpIG91IG8ocykgbm9tZXMocykgZG8ocykgZGV0ZW50b3IoZXMpIGRvcyBkaXJlaXRvcyBhdXRvcmFpcyBkYSBwdWJsaWNhw6fDo28sIGUgbsOjbyBmYXLDoSBxdWFscXVlciBhbHRlcmHDp8OjbywgYWzDqW0gZGFxdWVsYXMgY29uY2VkaWRhcyBwb3IgZXN0YSBsaWNlbsOnYS4KRepositório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-03-30T17:20:23Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
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