System biology as experimental strategy for discovery of novel natural products in the fauna of venomous arthropods from São Paulo State

Abstract

The demand for novel natural products is an important source of inspiration for most of the active ingredients of drugs worldwide; thus, there is a growing interest for the use of analytical platforms for increasingly sophisticated quantitative and qualitative identification of natural compounds in a fast and efficient way. Among these technologies, the combination of high resolution liquid chromatography and mass spectrometry of multiple steps (LC-MS and MSn) is the most used one. It is known so far that the insect venoms are complex mixtures of biologically active compounds, but few Brazilian species were chemically characterized to date. The main objective of this project is the standardization conditions for the analysis of the metabolomic "footprinting", using an LC (DAD) - IT-TOF/MS, MSn and GC-EI/HRMS, building initially, a database composed of low molecular weight compounds, characteristic of Arthropod venomous secretions. The venoms of different species of venomous Arthropods will be analyzed under the same analytical conditions in which the databank (DB) compounds, and the results compared with the DB for identification of low molecular weight compounds from such venomous secretions. This project will also address the development of protocols for studies of "analytical scanning" of natural library of peptides present in the venomous secretion of the Arthropods, based on the use of micro-scale techniques of LC-IT-TOF/MS MSn. The peptides sequenced by the strategy will be summarized and submitted to studies of characterization of the secondary structure of these peptides by spectroscopic methods (CD and NMR), and also to assays of biological activities, such as pain, inflammation, antibiosis, neurotoxicity / anti-epilepsy. The interaction of these peptides with membrane- mimetic systems (planar membranes and vesicles of different types) will be addressed with the use of biophysical techniques. Thus, this project seeks to fulfill some of the main objectives of the project BIOprospecTA -FAPESP for bioprospecting the biodiversity in the of São Paulo State. (AU)