Synthesis and structural investigation of peptides and polymers: from innovation in the synthesis method to the study of some physiological relevant peptides

• Grant number: 21/04885-3
• Support Opportunities: Regular Research Grants
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Clovis Ryuichi Nakaie
• Grantee: Clovis Ryuichi Nakaie
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated researchers: Adriana Karaoglanovic Carmona ; João Bosco Pesquero ; Luciana Malavolta Quaglio ; Maria Luiza Vilela Oliva

Abstract

Five sub-lines of research make up this project, a part strongly supported by improvements and technological innovations in the complex method of chemical synthesis of peptides (natural or not), carried out in polymers. This method, called solid-phase peptide synthesis (SPFS), winner of the 1984 Nobel Prize in Chemistry (Bruce R. Merrifield, Rockefeller University), has been investigated by us at different points in its experimental strategy, allowing us to For decades, we propose alternatives that have enabled not only the increase in the yield of peptide production but also the creation of innovative polymers and amino acid derivatives with application in this methodology. Also in the case of innovative polymers, we will test them under high-performance conditions and alternatively as solid supports for column chromatography. In addition to polymers, we will now focus on testing an alternative acid protocol in the critical step of cleavage of peptide chains from the polymeric support. This method makes use of the strong trifluoromethanesulfonic acid and has shown results that seem to indicate a proposal for a new peptide cleavage strategy in SPFS. At the end of this part of the project with a strong chemical connotation, we will advance in the study of the introduction of a new amino acid-type spin marker in the chemistry of peptides and polymers. It is the paramagnetic and fluorescence suppressor Poac (2,2,5,5-tetramethylpyrrolidine-N-oxyl-3-amino-4-carboxylic acid), which we will evaluate, compared to the one already introduced by us - Toac (acid 2, 2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxylic acid) in terms of feasibility of use. Unfortunately, this new spin marker has four types of enantiomers in its structure, which has made its synthesis and purification difficult. Due to these characteristics, we will try to find strategies that make its use viable. Just as an example of the social advantages in efforts and improvements achieved in the SPFS mentioned above, we have the case of the production with its own protocol, on a semi-industrial scale, of a therapeutic peptide that, after being transformed into a drug formulation (nasal solution), has been distributed to patients with Diabetes insipidus. This manufacturing stage and its distribution to the population, via SUS, was carried out by the Foundation for Popular Medicine (FURP), linked to the Health Department of the State of São Paulo in an agreement under my responsibility, with our University (Unifesp). This activity of extension and technological innovation of practical application is interrupted at the moment because in addition to having tochange some points in the synthesis protocol (explained later), the new managements of both Unifesp and FURP are in the process of trying to reach a legal agreement for the renewal of this process, which is relevant to the SUS. In addition, in the final two items, we intend to apply our learning in more biological themes. The first is related to the mechanism of action of the angiotensin I converting enzymes (ACE) and renin, both of the renin-angiotensin system (RAS). In the case of ACE, we will study the selectivity of this metalloendopeptidase through kinetic studies of several natural and unnatural analogues of the peptide (1-7)-Ang (DRVYIHP) which is a known inhibitor of the C domain of this enzyme. The challenge will be to find kinetic data that allow a better understanding of the mechanism of action of this enzyme, and that can also enable the synthesis of compounds with inhibitory activity for therapeutic purposes. A slightly different focus will be applied to the enzyme renin, studying, synthetically and conformationally, unnatural analogues of its tetradecapeptide substrate which releases angiotensin I in its hydrolytic action of selective endopeptidase. These are efforts to continue related works already developed, and special relevance will be given to derivatives containing Toac in differ (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

FERREIRA, MARIANA M. L.; DE SOUZA, SINVAL E. G.; DA SILVA, CAROLINE C.; SOUZA, LOUISE E. A.; BICEV, RENATA N.; DA SILVA, EMERSON R.; NAKAIE, CLOVIS R.. Pyroglutamination-Induced Changes in the Physicochemical Features of a CXCR4 Chemokine Peptide: Kinetic and Structural Analysis. BIOCHEMISTRY, v. 62, n. 17, p. 11-pg., 2023-08-04. (22/03056-6, 21/04885-3)

LOURENCO, THIAGO C.; DE MELLO, LUCAS R.; ICIMOTO, MARCELO Y.; BICEV, RENATA N.; HAMLEY, IAN W.; CASTELLETTO, VALERIA; NAKAIE, CLOVIS R.; DA SILVA, EMERSON R.. DNA-templated self-assembly of bradykinin into bioactive nanofibrils. SOFT MATTER, v. 19, n. 26, p. 11-pg., 2023-06-12. (21/04885-3, 22/03056-6, 19/19719-1, 21/10092-6)