Introduction: Artemisinin (1) is a milestone compound in malaria treatment, and it exhibits a broad scope of bioactivities. Herein, sequential chemo-reduction and biotransformation of artemisinin were undertaken to obtain a series of artemisinin derivatives.Methods: First, 10-deoxyartemisinin (2) and 9-ene-10-deoxyartemisinin (3) were synthesized after simple handling with boron trifluoride/diethyl ether and sodium borohydride. Then, biotransformation of 10-deoxyartemisinin was conducted with Cunninghamella echinulata CGMCC 3.4879 and Cunninghamella elegans CGMCC 3.4832, and the transformed products were separated and identified. The antimalarial activity of these products was tested in vitro against Plasmodium falciparum 3D7.Results: Fifteen metabolites (4–18), including seven novel compounds, were isolated and identified after cultivation. Compounds 2, 3, 13, 15, 16, and 18 displayed moderate-to-good antimalarial activity, with a half-maximal inhibitory concentration ranging from 6 to 223 nM.Discussion: This work explored the combination of chemical and biological transformation to develop a co-environmental, efficient, and cost-efficiency synthetic methodology and applied it to synthesize novel derivatives of artemisinin. The association of the two strategies will hopefully provide an abundant source for the development of novel drugs with bioactivities.

Dihydroartemisinin (DHA) is a sesquiterpene endoperoxide with prominent antimalarial efficacy, which was discovered by Professor Youyou Tu through the reduction of artemisinin in the 1970s. It is always a challenging work for scientists to investigate the metabolites of DHA in the red blood cells due to the complicated matrix background. As a bottleneck, the investigation of metabolites, especially exploring the pharmacodynamic material in the red blood cell, is necessary and significant for metabolism research of antimalarial agent. Recently, microbial transformation provides a green and economical means for mimicking mammal metabolism and synthesis active metabolites, based on which is one efficient route for drug discovery. In this study, a strain from Cunninghamella was employed as an efficient tool to explore active metabolites of DHA in erythrocyte. Microbial transformation products of DHA by Cunninghamella elegans CICC 40250 were detected and analyzed by ultra-performance liquid chromatography (UPLC)-electrospray ionization (ESI)-quadrupole time-of-flight (Q-TOF)-mass spectrometry (MS^E), and the main products were isolated and identified. The antimalarial activity of the isolated products was also screened in vitro. Totally, nine products were discovered through UPLC-ESI-QTOF-MS^E, and three main products with novel chemical structures were isolated for the first time, which were also detected in red blood cells as the metabolites of DHA. After evaluation, 7β-hydroxydihydroartemisinin (M1) exhibited a good antimalarial activity with an IC₅₀ value of 133 nM against Plasmodium falciparum (Pf.) 3D7. The structure and stereo-configuration of novel compound M1 were validated via X-ray single crystal diffraction. Microbial transformation was firstly employed as the appropriate model for metabolic simulation in erythrocyte of DHA. Three novel metabolites in erythrocyte were obtained for the first time through our microbial model, and one of which was found to show moderate antimalarial activity. This work provided a new research foundation for antimalarial drug discovery.

B(C₆F₅)₃-catalyzed cross-coupling reaction between allylic alcohols and boronic acids with excellent regioselectivity, broad scope and mild conditions.

A simple and effective method for the simultaneous quantitative analysis of six caffeoylquinic acids (CAs) in Matricaria chamomilla L. (M. chamomilla) using high-performance liquid chromatography (HPLC) with diode-array detection (DAD) was established. The chromatographic separation was performed on a Waters XBridge Shield RP C₁₈ column (4.6 mm × 250 mm, 5 μm) with the mobile phase of acetonitrile (0.5% phosphoric acid) and water (0.5% phosphoric acid) using a gradient elution at a flow rate of 1.0 mL/min and UV detection at 327 nm. The correlation coefficients of all analytes were 0.999, and the results showed excellent linearity. The lower limits of detection (LLOD) and quantification (LLOQ) of all analytes fall within the range of 0.014∼0.017 μg/mL and 0.068∼0.086 μg/mL, respectively. The extraction recoveries of all analytes fall within the range of 100.74%∼101.55%, with relative standard deviation not exceeding 2.83%. The intraday and interday precisions fall within the range of 0.03%∼0.65% and 0.02%∼0.09%, respectively. This validated method was successfully applied to the investigation of 34 samples of M. chamomilla collected from different geographical areas. The results showed that the established method is appropriate for the analysis of the six CAs in M. chamomilla and helpful for quality assessment of capitula of M. chamomilla (CMC), whole herb of M. chamomilla (WHMC), and related herbal formulas.

Background/Objectives: Medical leech (Hirudo in the Chinese Pharmacopoeia) is renowned in traditional medicine for its significant antithrombin activity. As an animal-derived medicine with complex and incompletely understood composition, its insufficient quality control measures are met with widespread counterfeiting caused by limited animal resources and rising demand. Methods: In this study, an integrated quality evaluation strategy guided by “Totality of the Evidence” (TOE) method is proposed. This strategy combines chemical characterization of small and macromolecular components with bioassays relevant to its clinical functions. A total of 28 batches of samples were analyzed, comprising 23 genuine and 5 counterfeit batches. Species origins were identified by morphology and DNA barcoding. Chemical characterization included TLC, HPLC and UPLC-QTOF-MS/MS for small molecules, and SDS-PAGE with HPLC-Orbitrap Fusion Lumos Tribrid-MS for macromolecules. Antithrombotic activity was assessed by thrombin titration and platelet aggregation assays. Results: Several characteristic components were discovered and identified as key quality control markers, including eight small molecules such as an unreported compound SZ-1, plus seven major differential proteins across species. Based on these markers, accurate and rapid authentication methods were established using SDS-PAGE for macromolecules, and both HPLC and TLC for small molecules. Furthermore, using bioassay methods we established for quality evaluation, Hirudo nipponica exhibits potent anti-thrombin activity and inhibits platelet aggregation, while Whitmania pigra shows weak anti-thrombin activity and promotes platelet aggregation. Conclusions: This quality evaluation strategy is not only applicable for the quality assessment of genuine Hirudo products of different origins, but also for distinguishing medical leeches from their counterfeits.

Toad Venom (TV) is the dried product of toxic secretions from Bufo bufo gargarizans Cantor (BgC) or B. melanostictus Schneider (BmS). Given the increasing medical demand and the severe depletion of wild toads, a number of counterfeit TVs appeared on the market, posing challenges to its quality control. In order to develop an efficient, feasible, and comprehensive approach to evaluate TV quality, a thorough analysis and comparison of chemical compounds among legal species BgC and BmS, as well as the main confusion species B. andrewsi Schmidt (BaS) and B. raddei Strauch (BrS), were conducted by ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS), high performance liquid chromatography (HPLC), sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and Nano LC-MS/MS analyses. We identified 126 compounds, including free or conjugated bufadienolides, indole alkaloids and amino acids, among the four Bufo species. The content of main bufadienolides, such as gamabufotalin, bufotalin, bufalin, cinobufagin, and resibufogenin, and the total protein contents varied widely among 28 batches of TV due to their origin species. The sum of the five bufadienolides within the BgC, BmS, BaS, and BrS samples were 8.15–15.93%, 2.45–4.14%, 11.15–13.50%, and 13.21–14.68%, respectively. The total protein content of BgC (6.9–24.4%) and BaS (19.1–20.6%) samples were higher than that of BmS (4.8–20.4%) and BrS (10.1–13.7%) samples. Additionally, a total of 1357 proteins were identified. There were differences between the protein compositions among the samples of the four Bufo species. The results indicated that BgC TV is of the highest quality; BaS and BrS TV could serve as alternative resources, whereas BmS TV performed poorly overall. This research provides evidence for developing approaches to evaluate TV quality and selecting the proper Bufo species as the origin source of TV listed in the Chinese pharmacopoeia.