Neuroprotective Effect of CR-777, a Glutathione Derivative of Withaferin A, Obtained through the Bioconversion of Withania somnifera (L.) Dunal Extract by the Fungus Beauveria bassiana.

The bioconversion of Withania somnifera extract by the fungus Beauveria bassiana leads to cysteine and glutathione derivatives of withaferin A at the C-6 position. The compounds were purified and fully characterized by 1D-NMR, 2D-NMR, and HRMS analysis. The glutathione derivative CR-777 was evaluated as a neuroprotective agent from damage caused by different neurotoxins mimicking molecular symptoms in Parkinson´s disease (PD), including 1-methyl-4-phenylpyridinium (MPP+), 6-hydroxydopamine (6-OHDA), and α-synuclein (α-Syn). CR-777, at nanomolar concentrations, protected dopaminergic and cortical neurons. In 6-OHDA-treated neurons, CR-777 increased cell survival and neurite network and decreased the expression of α-Syn. Using specific inhibitors of cell toxicity signaling pathways and specific staining experiments, the observed role of CR-777 seemed to involve the PI3K/mTOR pathway. CR-777 could be considered as a protective agent against a large panel of neuronal stressors and was engaged in further therapeutic development steps.

Antiangiogenic-Like Properties of Fermented Extracts of Ayurvedic Medicinal Plants.

The three ayurvedic medicinal plants, Withania somnifera, Emblica officinalis, and Bacopa monnieri, were extracted by high-pressure static extraction using the Zippertex(®) technology. The extracts were mixed to reach quantifiable amounts of active compounds identified by high-pressure liquid chromatography-mass spectrometry (HPLC-MS) analysis. The mixture of extracts was incubated with resting cells of the fungus Beauveria bassiana ATCC 7159. The fermentation promoted the fluidization of the starting dense mixture, while HPLC monitoring evidenced the disappearance of glucogallin from E. officinalis extract and the concomitant increase in gallic acid content. Topical exposure of the chick embryo chorioallantoic membrane (CAM) to the nonfermented extract led to the extensive necrosis and destruction of the treated membrane. However, the fermented extract was shown to be free of any toxicity. Furthermore, compared with the untreated CAM, the fermented sample reduced CAM vascularization, suggesting its antiangiogenic potency. The innocuity of the fermented extract was demonstrated using the in vivo LD50 test, the morphological examination of internal organs of treated rats, as well as the evaluation of blood biomarkers of liver damage (aspartate aminotransferase and alanine aminotransferase). The fermented extract was developed as a nutraceutical antiangiogenic treatment of age-related macular degeneration and commercialized in an oral form named Ethnodyne-Visio™.