Meeting the Challenge 2: Identification of Potential Chemical Probes for Parkinson’s Disease from Ligusticum chuanxiong Hort Using Cytological Profiling

Abstract

Traditional Chinese medicine (TCM) has been around for thousands of years and is increasingly gaining popularity in the Western world to treat various complex disorders including the incurable neurodegenerative condition, Parkinson’s Disease (PD). One of the many directions in recent studies of PD is utilizing the phenotypic assay, or cytological profiling, to evaluate the phenotypic changes of PD-implicated cellular components in patient-derived olfactory neuroepithelial (hONS) cells, upon treating the cells with extracts or pure compounds. To obtain small molecules for studies utilizing PD phenotyping assays, Ligusticum chuanxiong Hort was selected for analysis as it is a popular Chinese herbal medicine used for treating PD-like symptoms. Fifty-three secondary metabolites, including six new compounds, were isolated from the ethanolic extract of L. chuanxiong; their structures were elucidated based on several spectroscopic techniques such as NMR, MS, Fourier transform infrared (FTIR), UV, and theoretical density functional theory (DFT) calculations. Cytological profiling of the afforded natural products against PD hONS cells revealed 34 compounds strongly perturbated the staining of several cellular organelles. In fact, greaterthan 1.5-fold change was observed compared to the control (dimethyl sulfoxide; DMSO), with early endosome, lysosome, and autophagosome (LC3b) being particularly affected. Given these biological compartments are closely related to PD pathogenesis, the results helped rationalize the traditional medicinal use of L. chuanxiong in PD treatment. Further, the hit compounds can serve as chemical probes to map the molecular pathways underlying PD, potentially leading to new therapeutic targets for PD.

High-throughput discovery of highly selective reversible hMAO-B inhibitors based on at-line nanofractionation

“…Recently, Feng's groups adopted the cytological profiling (CP) technique of hONS cells to discover chemical probes from 24 TCMs and preparations against PD 23 . In Chuanxiong Rhizoma, mainly phenolic acids and phthalide compounds were identified as active ingredients against PD 31 . However, it was noticed that their anti-PD and neuroprotective targets have not yet been elucidated.…”

Isolation and In Vitro and In Vivo Activity of Secondary Metabolites from Clerodendrum polycephalum Baker against Plasmodium Malaria Parasites

“…The DFT calculation of the ECD spectra also followed the method described earlier . For each energy-minimized conformer, the ECD spectrum was computed at the B3LYP/TZVP level with 50 excited states using the IEF-PCM model for methanol.…”

“…The DFT calculation of the ECD spectra also followed the method described earlier. 60 For each energy-minimized conformer, the ECD spectrum was computed at the B3LYP/TZVP level with 50 excited states using the IEF-PCM model for methanol. The reported ECD spectra were the Boltzmann-weighted spectra of the optimized structures, which were generated using the equations by Stephens and Harada 61 and the single-point energies obtained above.…”