Mechanistic insights into honey-boiled detoxification of ChuanWu: a study on alkaloid transformation and supramolecular aggregation

Background : ChuanWu (CW), the dried mother root of Aconitum carmichaelii Debx., is a well-known traditional Chinese medicine (TCM) recognized for its potent efficacy and inherent toxicity. The alkaloid components present in CW are significant contributors to its toxic properties. Both traditional knowledge and modern advancements have facilitated the development of detoxification strategies for CW, which include appropriate processing, rational compatibility, and specialized decoction methods. Among these approaches, honey-boiled CW emerges as a distinctive detoxification technique. However, research on the detoxification mechanism of honey- boiled CW remains limited.
Aim of the study : This study aimed to investigate the detoxification mechanism of honey-boiled CW by examining alkaloid transformation and supramolecular aggregation.
Materials and methods : Honey-boiled CW and water-boiled CW were prepared separately for comparative analysis. Ultra-high-performance liquid chromatography- tandem mass spectrometry （ UHPLC-MS/MS） was employed to analyze CW alkaloids, specifically diester alkaloids (DDAs), monoester alkaloids (MDAs), and non- 18 esterified diterpenoid alkaloids (NDAs). Transmission electron microscopy (TEM) was utilized to observe supramolecular aggregation in the honey-boiled CW decoction, which was subsequently isolated and chemically identified. Assemblies of natural deep eutectic solution (NADES) and monomers nanoparticles were prepared for pharmacokinetic studies after oral administration to rats. In addition, we compared the in vivo absorption differences between water-boiled CW, honey-boiled CW and NADES-boiled CW. The toxicity of various treatments for CW was assessed using the LD₅₀ test, with evaluations of both hepatotoxicity and nephrotoxicity. The safety, as well as the anti-inflammatory and analgesic effects of different treatments of CW medicated serum on RAW264.7 cells, were examined through in vitro experiments. These anti-inflammatory and analgesic effects were then further validated through in vivo experiments conducted on mice. Journal Pre-proof4.
Results : Honey played a significant role in altering the alkaloids in boiled CW by promoting the conversion of highly toxic DDAs to less toxic MDAs and preventing the 3 hydrolysis of MDAs into NDAs. Additionally, the honey-boiled CW facilitated the formation of supramolecular aggregates with a diameter of approximately 250 nm. These aggregates effectively hindered the conversion of MDAs into NDAs by encapsulating them. The encapsulated MDAs within the supramolecular aggregates served as a stable drug delivery system under physiological conditions and demonstrated higher bioavailability compared to free benzoylmesaconine (BMA). Subsequent mouse experiments confirmed that honey-boiled CW significantly increased LD₅₀ of CW while reducing hepatotoxicity and nephrotoxicity. Honey-boiled CW significantly improves cell safety and enhances anti-inflammatory and analgesic effects.
Conclusions : Honey-boiled CW exhibited a potent detoxification mechanism by influencing alkaloid transformation and facilitating the formation of supramolecular aggregates. This finding lays the groundwork for the development of detoxification or synergistic strategies within honey-boiled TCM.