Studies On The Chemical Constituents And Assembling Functional Activity Of Two Kinds Of Chinese Medicinal Herb Based On The Supramolecular Structures

Supramolecular structures refer to the spontaneous assembly and association of the same or different molecules through various non-covalent interactions to form a multimolecular aggregate structures having clear microstructure and macro phase state in the size range from nano to micrometer,whose main functions include molecular recognition,material transport,etc.It has been reported that some chemical components in Chinese medicinal herb had excellent supramolecular assembly performance.The interpretation of the active substances of Chinese medicinal herb based on chemical components forming supramolecular structures has become the frontier and hot research direction at home and abroad.On the basis of summarizing a large number of relevant literature and previous work,we have proposed a theoretical hypothesis for the assembly of active ingredients in Chinese medicinal herb,that was,Chinese medicinal herb could be regarded as a multicomponent system,in which there were a considerable proportion and quantity of natural small molecules with self-assembly properties,which could spontaneously deliver other active ingredients or co-assemble with them through various non-covalent forces,enabling the extracts to exert overall efficacy in the macroscopic form of supramolecular aggregates.Through combining emerging supramolecular assembly chemistry methods with traditional natural product separation technologies,the chemical components and their assembly modes,functional bioactivities of two kinds of Chinese medicinal herb,the roots of Rosa laevigata and the seeds of Orychophragmus violaceus,had been studied systematically.Based on supramolecular structures,we hope to interpret deeply the the bioactive substantial basis of Chinese medicinal herb for its further promotion and application.1.Studies on chemical constituents and assembly functional activities of the roots of R.Laevigata based on supramolecular structuresTwenty known compounds(Al-20),including eighteen triterpenes,one steroid,and one flavonoid,were isolated from the roots of R.laevigata by chromatography techniques.In the process of extraction and separation,several self-assembled natural product gelators(NPGs)were found by the vial inversion method,among which the pomoic acid(PA,A8)and fupenzic acid(FA,A7)were both novel self-assembled NPGs.It was found that PA could be induced to form gel by heating and cooling,while FA could be directly induced to form gel by heating,which was also the first heat-induced NPG.The micromorphology of them were observed by SEM,TEM,and AFM,and their rheological properties were characterized by various rheological techniques.A series of spectroscopic methods(IR,UV,NMR,etc.)and MD simulations had confirmed that non-covalent interactions such as hydrogen bonds,van der Waals forces,and hydrophobic interactions were the main driving forces for their molecular self-assembly.Comparing with free PA,it was preliminarily found that PA-gel exhibited stronger cytotoxicity against human breast cancer(MCF-7)cells in vitro by MTT method.For FA-gel,by establishing a bearing human malignant glioma cell(U118-MG)nude mouse model,the anti-tumor activity of it was evaluated,which proved that the tumor inhibitory activity and biological safety of FA-gel were better than those of free FA.2.Studies on chemical constituents and assembly functional activities of the seeds of O.violaceus based on supramolecular structuresIn the systematic chemical composition study of the seeds of O.violaceus,sixty compounds(B1-60)were isolated and identified,including twenty-two alkaloids,twentysix phenolic acids(glycosides),seven flavonoids,and five others.There were eight new compounds(B1-8),named as Orychophragvioline A-F(B1-6),Orychophragvioside A-B(B7-8),and compound B1 had a unique framework of 1-benzene-2,3-diketopiperazine.In addition,it was found that adenosine(Ado,B20)could self-assemble to form supramolecular hydrogels(Ado-gel)by heating and cooling for the first time,and also coassemble with gentisic acid(DHB,B40)from O.violaceus to form hydrogels(DA-gel).It was found that the micromorphology of DA-gel was different significantly from that of the monomers via various electron microscopic observations.By comparing the rheological parameters,it was suggested that the rheological properties of the DA-gel were better than that of the Ado-gel.And then,SAXS,DSC,contact angle measurements,etc.were used to conduct in-depth physical characterization of the DA-gel.The complete process and interaction forces of the two component co-assembly were analyzed using Gaussian theoretical calculations and MD simulation(all atom simulation+coarse particle simulation).The simulation results were verified by UV,IR,XRD,NMR,fluorescence spectroscopy,and other experimental methods.It was proved that the spontaneous coassembly effect between the two components mainly through hydrogen bonding.π-πstacking interaction,supplemented by van der Waals force,and electrostatic interaction.In further studies of functional biological activities in vitro,DPPH clearance and total reductive capacity experiments,LPS-induced RAW 264.7 cell inflammation model,and oxygen and glucose deprived rat cardiac H9C2 cell model were used to demonstrate that the antioxidant,anti-inflammatory,and myocardial cell protection activities of the two components were superior after co-assembly to those of the free monomer components and non assembly mixed components,suggesting that the functional biological activities of the two components were significantly improved after co-assembly.3.Research resultsThis study had innovatively integrated the theory and technology of supramolecular assembly chemistry into the research process of natural products.Two new natural selfassembled small molecules were discovered from twenty known compounds isolated from the roots of R.laevigata.The self-assembly mechanism was explored through MD simulation and experimental measurements.It was found that their anti-tumor activities had been increased after self-assembly in vitro and in vivo.Sixty compounds including eight new compounds(containing a new skeleton compound)were isolated from the seeds of D.violaceus.At the same time,a novel adenosine-gentianic acid co-assembled hydrogel(DA-gel)was found and prepared,and its co-assembly mechanism was systematically explored.Through in vitro experiments,it was found that the co-assembled hydrogel had excellent antioxidant,anti-inflammatory and myocardial protection activities.The research on chemical components of two kinds of Chinese medicinal herb based on supramolecular structures aims to preliminarily verify the " theoretical hypothesis of active assembly components of Chinese medicinal herb ".Combining theory with practice,it have provided new insights and ideas for interpreting the action mode of intrinsic active substances of Chinese medicinal herb,and also laid a foundation for the research and application development of the action mechanism of active natural products.