Bio-guided isolation of new natural anti-venom agents from extracts reported on the work of the ancient Greek physician, Nicander

Nicander was a famous poet-naturalist of the 2nd century BC. Theriaca is his second surviving work, a Hellenistic poem, on the bites of venomous creatures, while a series of herbal remedies are also mentioned. The aim of this study was to evaluate the chemical content and biological properties of plants reported on Theriaca and in particular to investigate the possibility of their application as therapeutic agents for wound healing and neurodegenerative diseases. Plants were collected, dried and extracted by Accelerated Solvent Extraction (ASE) technique resulting in the creation of a 'theriaca extracts' library. Their chemical profile was analyzed by HPTLC and LC-MS. The anti-venom potent of the most promising extracts was evaluated by enzymatic inhibitory assays. Hyaluronidase, phospholipase-A2 and acetylcholinesterase [1] are common molecular targets for the evaluation of anti-ophidian properties of natural products. Paeonia parnassica, a Greek endemic species, and three Aristolochia spp. were chosen for fractionation and separation of their chemical constituents by means of Centrifungal Partition Chromatography. The obtained fractions were evaluated for their ability to inhibit the enzyme targets where the initial extract was active. In parallel the chemical composition of each fraction was established by NMR-dereplication approach and analyzed by HPTLC. Finally, five anti-phospholipase agents corresponding to aristolochic acid derivatives from Aristolochia spp. and ten anti-acetylcholinesterase and anti-hyaluronidase components belonging to phenolics and flavonoids from Paeonia parnassica were identified.

[1] Molander M, Nielsen L, Søgaard S, Staerk D, Rønsted N, Diallo D, Chifundera KZ, Staden J, Jäger AK. J Ethnopharmacology 2014; 18:157:171 – 80