Bioaffinity Fishing Procedure Using Secretory Phospholipase A2 for Screening for Bioactive Components: Modulation of Pharmacological Effect Induced by sPLA2 from Crotalus durissus terrificus by Hispidulin from Moquiniastrum floribundum

Abstract

Bioaffinity capturing of molecules allows the discovery of bioactive compounds and decreases the need for various stages in the natural compound isolation process. Despite the high selectivity of this technique, the screening and identification methodology depends on the presence of a protein to capture potential ligands. However, some proteins, such as snake secretory phospholipase A2 (sPLA2), have never been investigated using this approach. The purpose of this study was to evaluate the use of a new method for screening natural compounds using a bioaffinity-guided ultrafiltration method on Crotalus durissus terrificus sPLA2 followed by HPLC-MS to identify the compounds, and this method could be used to discover new anti-inflammatory compounds from the various organisms originating from biodiversity. Different extracts were selected to evaluate their ability to inhibit sPLA2 activity. The extracts were incubated with sPLA2 and the resulting mixture was ultrafiltrated to elute unbound components. The resulting compounds were identified by HPLC-MS. We identified hispidulin as one of the components present in the Moquiniastrum floribundum leaf and evaluated the ability of this isolated compound to neutralize the inflammatory activity of sPLA2 from Crotalus durissus terrificus.

Affinity selection–mass spectrometry for the discovery of pharmacologically active compounds from combinatorial libraries and natural products

“…PUF AS‐MS has been applied to the discovery of ligands to a wide variety of pharmacological targets, including some that are not amenable to conventional HTS. Anti‐inflammation targets have included cyclooxygenase‐1 23 and cyclooxygenase‐2, 24,25 secretory phospholipase A2, 26 and 5‐lipoxygenase 27 . Parkinson's disease and Alzheimer's disease targets have included tyrosinase 28 and acetylcholinesterase 29 .…”

Editorial to Special Issue—Anti-Inflammatory Activity of Natural Products

“…Secretory A2 phospholipases (sPLA2) is a pro-inflammatory protein involved in the mobilization of arachidonic acid by an indirect activation of cytosolic phospholipase A2 (cPLA2) and phospholipase C (PLC). sPLA2 from snake (Crotalus durissus terrificus) venom has structural similarity to that of human origin whereby Santos Júnior [19] used a bioaffinity-guided ultrafiltration method on Crotalus durissus terrificus sPLA2 in order to detect new anti-inflammatory compounds of diverse origin. In this case, the authors [19] used compounds isolated from Moquiniastrum floribundum (Asteraceae) leaf for evaluating their ability for neutralizing the inflammatory activity of sPLA2 from Crotalus durissus terrificus.…”

“…sPLA2 from snake (Crotalus durissus terrificus) venom has structural similarity to that of human origin whereby Santos Júnior [19] used a bioaffinity-guided ultrafiltration method on Crotalus durissus terrificus sPLA2 in order to detect new anti-inflammatory compounds of diverse origin. In this case, the authors [19] used compounds isolated from Moquiniastrum floribundum (Asteraceae) leaf for evaluating their ability for neutralizing the inflammatory activity of sPLA2 from Crotalus durissus terrificus. From diverse extracts using organic solvents, the methanol one was that better inhibit sPLA2.…”

“…From diverse extracts using organic solvents, the methanol one was that better inhibit sPLA2. Such extract had various compounds but the approach (bioaffinity technique) followed by the authors permitted them to conclude that the flavone hispidulin was able to inhibit the edematogenic and myotoxic activity of sPLA2 but not a decrease in sPLA2 [19].…”