| As a commonly used of traditional Chinese medicine,the Periostracum Cicadae is contained in the "Medical Doctors",which is included in the "Chinese Pharmacopoeia" 2015 edition,and is the shell of the cicada Cryptotympana pustulata Fabricius that is cast-off during molting,has the functions of dispelling pathogenic wind,relieving fever,relieving pharynx,removing blood stasis,penetrating rash and relieving spasmolysis,Commonly used in pharyngitis,nephritis,headache,nocturnal crowing in children and other diseases.Ganderma lucidum(Leyss.Ex Fr.)Karst is a main variety of Ganoderma listed in "Chinese Pharmacopoeia" 2015 edition,which has the effects of strengthening body resistance,consolidating the body,and prolonging life,It has significant curative effect on heart disease,hepatopulmonary disease,cerebrovascular disease and so on.For a long time,macromolecules such as proteins and amino acids have been considered as the main active components of Periostracum Cicadae.Is there a novel structure with small molecules? What is its possible biological significance? still not clear.The chemical constituents of Ganoderma lucidum are mainly polysaccharides,triterpenoids,etc.In recent years,the miscellaneous components in Ganoderma lucidum have also attracted the attention of some research institutions and achieved certain results.In this paper,we will study the small molecular compounds in Periostracum Cicadae and the heteroterpenoids in Ganoderma lucidum,which will help people to recognize the structural diversity of the active components of Periostracum Cicadae and Ganoderma lucidum,and also provide an important template molecules for the development of biosynthesis;The prediction of anti-tumor molecular docking activity of new compounds isolated and identified from Periostracum Cicadae and Ganoderma lucidum can provide a scientific basis for clarifying the pharmacodynamic substance basis,action mechanism and quality standard of Periostracum Cicadae and Ganoderma lucidum.Objective1.The chemical constituents were separated from the water part of Periostracum Cicadae and the ethyl acetate part of Ganoderma lucidum after extraction,and the structure of monomers were identified by modern spectroscopic techniques.To reveal the chemical constituents contained in Periostracum Cicadae and Ganoderma lucidum.2.To predict the anti-tumor molecular docking activity of the new compounds isolated and identified from Periostracum Cicadae and Ganoderma lucidum in order to obtain the chemical constituents with good biological activity in Periostracum Cicadae and Ganoderma lucidum,which was the basis of anti-tumor activity experimental identification in the later stage of Periostracum Cicadae and Ganoderma lucidum.Methods1.The dried Periostracum Cicadae was extracted by methanol / water(100:0)and ethanol / water(80:20)at room temperature,the extracts were combined to concentrate the Periostracum Cicadae extract.The dried Ganoderma lucidum extractwas extracted by ethanol / water(95:5)percolation at room temperature.The extract was concentrated and extracted with ethyl acetate for 3 times to obtain ethyl acetate fraction.2.The traditional and modern separation techniques such as MCI gel CHP 20 P,Sephadex LH-20,RP-18,normal phase silica gel,preparation / semi-preparation type high performance liquid chromatography(HPLC)and thin layer chromatography(TLC)were used for the obtained bismuth extract and ethyl acetate.Separation and purification are repeated to finally obtain a monomer compound.The structures of the isolated and purified compounds were analyzed and identified by 1D/2D NMR,MS,HRMS,UV,CD and other spectroscopy methods.3.Maestro docking technique was used to analyze the interaction of ten new chemical constituents isolated from Periostracum Cicadae and Ganoderma lucidum in anti-tumor related targets and to predict the molecular mechanism of the new compounds in anti-tumor.Results1.A total of 29 compounds were isolated and identified from the above two Chinese herbs,of which 20 were from Periostracum Cicadae and 9 were from Ganoderma lucidum.11 known compounds in Periostracum Cicadae were N-(2-(3?,4?-dihydroxybenzoyl)-4,5-dihydroxyphenethyl)acetamide(10),trans-2-(3?,4?-Dihydroxyphenyl)-3-acetylamino-6-(1??,2??-dihydroxyethyl)-1,4-benzodioxane(11),trans-2-(3?,4?-Dihydroxyphenyl)-3-ace-tylamino-6-acetyl-2??-aminoethyl-ene)-1,4-ben zodioxane(12),trans-2-(3?,4?-Dih-ydroxyphenyl)-3-acetylamino-6-hydroxyethyl-1,4-benzodioxane(13),trans-2-(3?,4?-Dihydroxyphenyl)-3-acetylamino-6-(N-acetyl-1??-hydroxyl-2??-aminoethyl)-1,4-benzodioxane(14),cis-Cyclo(L-4-hydroxypro-L-Phe)(15),3-Hydroxy-9-methoxypterocarpan(16),3-Hydroxy-8,9-methylenedioxypterocar pan(17),2-(3",4"-dihydroxyphenyl)-1"-oxoethylacetate(18),4-Hydroxy-3-methoxy-benzoic acid(19),4-Hydroxybenzaldehyde(20);8 known compounds in Ganoderma lucidum were 6?-acetyl-4-hydroxy-1-oxo-3H-spiro[benzofuran-2?,3?-cyclopentane]-9?-carboxylic acid(2),4-hydroxy-6?-(9?-hydroxyprop-7?-en-8?-yl)-1-oxo-3H-spiro[benzo furan-2?,3?-cyclopentane]-10?-carboxylicacid(3),4-hydroxy-6?-(8?-hydroxypropan-7?-yl)-1-oxo-3H-spiro[benzofuran-2?,3?-cyclopen-tane]-10?-carboxylic acid(4),1?-(2-(1,4-dihydroxyphenyl)-2-oxoethyl)-6?-hydroxytet-rahydro-1H,3H-cyclopenta[1?,2?-b:7?,8?-c?]difuran-3?,7?(8H)-dione(5),3,9-dihydroxychr-omeno[4,3-c]chromene-5,11-dione(6),1-oxo-1,3-dihydroisobenzofuran-5-carboxylic acid(7),6-hydroxy-2H-chromen-2-one(8),methyl 2,5-dihydroxybenzoate(9)?2.Nine new compounds were isolated from Periostracum Cicadae,named cicadamide A(1),cicadamide B(2),cicadamide C(3),cicadamide D(4),cicadamide E(5),cicadamide F(6),cicadamide G(7),cicadamide H(8),cicadamide I(9);One new compounds were isolated from Ganoderma lucidum,named Spiro-lingzhine A(1).3.By docking new compounds in Periostracum Cicadae and Ganoderma lucidum mollusk with anti-tumor target molecules,6 targets such as ESR1,TP53,MYC and so on were found to have better docking with new compound 1.There are14 targets,such as ESR1,VEGFA and PIK3 CA were found to have good docking with new compound 2,10 targets such as ESR1,PIK3 CA and MYC were found to have good docking with new compound 3,9 targets such as EGFR,PIK3 CA and TP53 were found to have good docking with new compound 4,9 targets such as ESR1,MYC and KRAS were found to have good docking with new compound 5,9targets such as MYC,PIK3 CA and ERBB2 were found to have good docking with new compound 6;17 targets such as ESR1,ERBB2 and MYC were found to have good docking with new compound 7;13 targets such as ESR1,EGFR and MYC were found to have good docking with new compound 8;11 targets such as MYC,ESR1 and VEGFA were better docked with new compound 9;8 targets such as ESR1,ERBB2 and PIK3 CA were found to have good docking with new compound 10.According to statistics,the protein targets ERBB2,MYC and PIK3 CA have good docking affinity with 10 new compounds,and the overall score of ERBB2 is higher in the scoring list,which proves that it has a stronger affinity to each compound.Conclusion1.By studying the small molecular compounds in Periostracum Cicadae,9 new compounds were isolated and identified,which provided a basis for the study of the material basis of Periostracum Cicadae and also provided a reference for the study of insect small molecular compounds.2.In this paper,a new framework of heteroterpenes was isolated and identified by focusing on the studies on the components of Heteroterpenes in Ganoderma lucidum,which provided a scientific basis for the study of the pharmacodynamic material basis of Ganoderma lucidum.3.The interaction of ten new chemical constituents isolated from Periostracum Cicadae and Ganoderma lucidum on anti-tumor-related targets was analyzed by Maestro molecular docking technique.The results showed that protein target ERBB2,MYC and PIK3 CA had good docking affinity with ten new compounds,respectively.Ten compounds may be active in anti-breast cancer,esophageal cancer and so on. |
PDF Full Text Request