Natural products represent a traditionally very important source of novel drugs and also a relevant inspiration for the synthesis of novel molecules of pharmaceutical interest. Recently, interest in natural bioactive compounds has raised, together with the need to identify new environmentally friendly extraction methods to obtain purified bioactive molecules. In this framework, our aim is to study the capacity of natural deep eutectic solvents (NaDES) - which are emerging as new green solvents able to replace current harsh organic solvents ¿ to extract polyphenols from plants. Polyphenols are well known antioxidant, antiviral, anti-inflammatory and neuroprotective compounds, hence these extracts will be tested for their capacity to counteract the neurodegenerative effects of Beta Amyloid (Ab) in cellular and animal models of Alzheimer¿s disease (AD).
As raw materials we will use various ecotypes of Arabidopsis thaliana, a plant of the Brassicaceae family, known for its high content of bioactive molecules such as polyphenols, glucosinolates, carotenoids and vitamins. After an initial characterization of the extracts by chromatographic and spectroscopic methods, the extracts will be tested in cellular models of neurodegeneration, i.e. neuronal (SH-SY5Y) or microglial (BV2) cells to assess antioxidant and anti-inflammatory capacity, respectively. The extracts will also be tested in a Drosophila model of AD, to evaluate effects on lifespan and on climbing ability of the flies. The extracts will also be tested for their antiviral activity in a cellular model of HPV infection.
We consider this project innovative because it will offer a new way of obtaining extracts enriched in antioxidants, antiviral and neuroprotective substances, without the use of harmful organic solvents or other harsh systems. Furthermore, as the NaDES are composed of edible molecules, the extract are ready-to-use, biocompatible and eco-friendly. The absence of any known toxicity of the principal constituents of NaDES (sugars, aminoacids, organic acids) which are normal components of food or arise from metabolic reactions in our body, makes these solvents extremely promising to the development of new food supplements/nutraceuticals enriched in protective substances.
Furthermore, clarifying the effects of complex mixtures of polyphenols with possible synergistic activity may shed new light on the role nutritional therapy in neurodegenerative disease. The detailed chromatographic characterization of the extracts will also allow a correlation with the biological activity in order to define a fingerprint of bioactive components in complex mixtures.
The efficacy of the extracts will be assessed in two different models of disease: a neurodegenerative diseases such as AD and an infectious disease such as the Papilloma virus infection.
Disclosing the molecular mechanism of polyphenol action may also provide insights into disease pathway and contribute to identify new pharmacological targets and possible alternative therapeutic interventions. The project involves two different research groups whose scientific expertise spans from biochemistry to molecular biology to microbiology. They will cooperate to develop this project in a highly synergistic way, each providing its own specific sound experience and capacity, and sharing a series of different techniques and experimental settings to obtain fully integrated information on the therapeutic potential of polyphenols in the treatment of AD. All the methods and techniques are already available and in current use. No major technical pitfalls are expected.