Quality And Variance-types Study On Panax Quinquefolius And Screening Of EGFR-TKI From 49 Herbs

Panax quinquefolius belongs to the Panax genius, Araliaceae family. P. quinquefolius, also named as American ginseng is native to eastern North America, and grows as an understory plant in deciduous and mixed forests in regions of the northeastern states of the United States (Wisconsin) and the Canadian provinces of Quebec and Ontario.Changbai Mountain Area of Jilin province, Weihai area of Shandong province and Liuba Hanzhong area of Shanxi province in China are also the major areas for cultivating the P. quinquefolius.The herbal is recorded in Chinese Pharmacopoeia(2015 version). In this study, quantitative determination of the main components, multivariate statistical analysis and DNA barcoding were used for researching the relationship among the quality, chemotypes and genetic of P. quinquefolius.P. quinquefolius is a very expensive herbal medicine and it has been widely used as medicines and dietary supplements in China and North America. Other studies have showed that the main active compounds are ginsenosides. According to 2015 CP, the content levels of Rg1, Re and Rbl were the quality control standard for P. quinquefolius. In the study, a HPLC-UV method was established for analyzing the contents of 7 ginsenosides and it will be more significanct for evaluating the quality of it.69 batches of the P. quinquefolius from 4 different areas were analyzed by the method. The quality of P. quinquefolius from Jilin and Weihai was better than that of Wisconsin and Shanxi. Principal component analysis(PCA) and Hierarchical cluster(Ward method, caculated by Euclidean distance) were used to analyze the difference of the P. quinquefolius among the 4 areas. Jilin, Weihai of Shandong and Wisconsin cluster a group and Hanzhong cluster an independent group. Dongbei, Weihai of Shandong and Wisconsin were defined as the low content of comA and comB type and Hanzhong of Shanxi was defined as high content of comA and comB type. There are two chemtypes based on the content of Ginsenoside A and B. The psbA-trnH sequences of 69 P. quinquefolius were analyzed for intraspecific variation. There were not intraspecific variations. It can concluded that the psbA-trnH can not identify the samples from different origins. The psbA-trnH was also used to identify the different species in Ginseng genus. And the species can be divided from P. quinquefolius except the P. ginseng.Epidermal growth factor receptor (EGFR) is a validated target in different human malignancies. EGFR tyrosine kinase inhibitors are known to contribute considerably to the extension of progression-free survival in EGFR-mutant non-small cell lung cancer. Monoclonal antibodies targeting EGFR have also improved the efficacy outcomes in KRAS wild-type colorectal cancer. At present, gefitinib and erlotinib are considered to be the most representative drugs approved by the FDA for cancer treatment. However, only a few studies have screened the active compounds of traditional Chinese medicines.In this study, the EGFR inhibitory activities of extracts of Chinese medicinal herbs that are traditionally prescribed to treat cancer were examined in a 384-well plate assay based on homogeneous time-resolved fluoroimmunoassay method. Petroleum ether, ethanol, ethyl acetate, and aqueous extracts of 49 traditional Chinese medicinal herbs (total 183 extracts) were tested. The inhibitory effects were expressed as percentage of inhibition. Dose-dependent inhibitory assays were further performed for the herbal extracts found to exert significant inhibition. Results The ethyl acetate extracts of Artemisia argyi, Lonicera macranthoides, Spatholobus suberectus, Curcuma longa, and Galla Chinensis, as well as the ethanol extract of Eriobotrya japonica exhibited remarkable EGFR inhibitory activities at the final concentration of 50μg/ml. Their IC50 values were 8.848,2.027,2.277,3.621,4.339 and 5.528μg/mL, respectively. The EA extracts of Lonicera macranthoides, Spatholobus suberectus show the best activities.Our results indicate that the screening of traditional Chinese herbs for EGFR inhibitory activity may provide considerable potential lead compounds in the discovery of new drugs for the treatment of cancer...