Design,Synthesis And Anti-Inflammatory Effects Of Cannabinoid Compounds

Cannabidiol（CBD）is a naturally occurring terpenoid in plants.Due to its neuroprotective,antidepressant,anxiolytic,antiepileptic,anti-inflammatory,analgesic and other pharmacological effects,CBD has attracted much attention from scholars at home and abroad in recent years.However,the disadvantages of CBD,such as poor water solubility,low bioavailability and nerve cell toxicity,have severely limited its further pharmacological applications.In this thesis,two strategies,pharmacochemical and modulation by bio-orthogonal reactions,will be used in anticipation of eliminating or attenuating its nerve cell toxicity and improving the therapeutic window.The specific studies are as follows:In the first part of the work,a series of CBD derivatives were designed and synthesized by organic chemistry/pharmaceutical chemistry methods,focusing on two key sites of the terpene ring and the long chain of the phenol side chain in the CBD structure,which were modified in four different aspects,namely,the partial structure of the CBD terpene ring,the benzyl position of the side chain of the CBD benzene ring,the structure of the CBD benzene ring and the derivative synthesis of divalent CBD.The structures of the synthesized derivatives were characterized and confirmed by modern wave spectroscopy techniques(¹H NMR,¹³C NMR).On this basis,the compounds were evaluated for their inhibitory effect on NO inflammation model induced by lipopolysaccharide（LPS）using BV-2 cells as a model.Two compounds 11 and 16 with comparable activity to CBD were obtained by screening,with IC₅₀of 18.65μM and 19.16μM,respectively,and their toxicity was also weaker than that of CBD.finally,the conformational relationship between cannabidiol derivatives and anti-inflammatory activity was preliminarily elucidated.The second part is to eliminate the nerve cell toxicity of CBD using a switching strategy.The phenolic hydroxyl group is the key group for CBD to function,and the bio-orthogonal bond-breaking reaction of tetrazine compounds with vinyl ethers was used to protect and deprotect the phenolic hydroxyl group of CBD,thus regulating its activity and toxicity.Firstly,the synthesis of CBD phenolic hydroxyl modification probes was performed by introducing one or two vinyl protecting groups at the hydroxyl position of CBD to obtain probes 3-2 and 3-3,and the disappearance of both anti-inflammatory activity and nerve cell toxicity of CBD after masking of its hydroxyl group was demonstrated using BV-2 cells.Subsequent in vitro experiments using bioorthogonal reactions with tetrazines successfully removed the introduced vinyl protecting group,demonstrating the feasibility of this strategy.Meanwhile,in order to improve the reaction rate of tetrazine with vinyl ether CBD derivatives,different kinds of tetrazine compounds were screened in this part,and finally the most excellent tetrazine compound,dimethyl 1,2,4,5-tetrazine-3,6-dicarboxylate,was found.The reaction kinetics of tetrazines with vinyl ether CBD derivatives were monitored by liquid chromatography.Using BV-2 neuroglia,the cells were collected at different time periods after drug administration,and the drug standard curves were established by HPLC/UV,and the drug content of CBD,probe 3-2 and probe 3-3 was tested through the cell membrane into the cells,and the cellular transport process of CBD analogues was found to be closely related to the phenolic hydroxyl group in the structure.In this thesis,a series of cannabidiol derivatives were synthesized and their anti-inflammatory pharmacological activities were investigated to preliminarily elucidate the relationship between the structure and anti-inflammatory activity of cannabidiol derivatives,which provides ideas for the design of novel anti-inflammatory drug lead compounds.In addition,the bio-orthogonal reaction strategy employed to control the activity switch of modified CBD verified its feasibility,wide applicability and possible applicability for in vivo applications.