â… . Chemical Constituents From Endophytic Fungus Periconia Sp. F-31Isolated From Annona Muricataâ…¡. Liposoluble Chemical Constituents From Cell Cultures Of Salvia Miltiorrhiza

1. Chemical constituents from endophytic fungus Periconia sp. F-31isolated from Annona muricataPericonia sp. F-31was an endophytic fungus isolated from the medicinal plant Annona muricata, which might be a new species according to the identification by using morphological and molecular (ITS1-5.8S-ITS2rDNA sequence) analyses, the crude extract of which displayed potent cytotoxicity against several human cancer cell lines.As part of our search for novel bio-active PKS-NRPS metabolites from Periconia sp. F-31, the EtOAc extract of the filtrate and mycelia of this strain were systematically investigated. Under the guidance of HPLC-UV-MS" analysis, twenty-nine compounds were isolated by using various chromatographic techinques, and their structures were elucidated by extensive spectroscopic data analysis. Especially, the absolute configurations were determined by analyzing the single-crystal X-ray diffraction data, CD [including Rh2(OCOCF3)4-induced CD^Mo2(OAC)4-induced CD] spectroscopic data. Twenty-four (P1-P24) of them are new compounds, including sixteen PKS-NRPS metabolites (P1-P16), these PKS-NRPS metabolites contain eight novel cytochalasans (P1-P8) and two PKS-NRPS metabolites with an unique6/6/5tricyclic ring system (P9-P10). The cytochalasan contain three type structures:the first type of which possesses novel9/6/5tricyclic ring system, the second type has unique5/6/6/5and5/5/6/6/5carbocyclic ring system which might be derived from the former, the third type displays unusual7/6/5tricyclic ring system. The plausible biosynthetic pathways of new compounds were also proposed.The in vitro bio-activities of part of compounds were evaluated for the cytotoxicity against five cancer cell lines and antiviral activity. Comopunds PI and P2showed selective and significant cytotoxicity against HCT-8, BGC-823, and Bel-7402cancer cell lines. The discovery of novel PKS-NRPS metabolites with cytotoxicities not only provide the diverse molecules for lead compounds, but also are useful for further biosynthesis investigation of these significant PKS-NRPS metabolites. 2. Liposoluble chemical constituents from cell cultures of Salvia miltiorrhizaSalvia miltiorrhiza Bunge (Labiatae), which is well-known as the traditional Chinese medicine "Danshen", has been used for the treatment of menstrual disorders, menorrhagia, menostasis, insomnia and cardiovascular diseases, particularly angina pectoris and myocardial infarction. The liposoluble chemical constituents of S. miltiorrhiza were confirmed as effective constituents, which were mainly used for the treatment of coronary heart disease. Numerous studies of chemical constituents of S. miltiorrhiza plants have been reported, however, the chemical constituents of the cell cultures of S. miltiorrhiza have hardly reported till now. As part of our search for bioactive compounds, the petroleum ether fraction of80%EtOH extract of S. miltiorrhiza cell cultures were systematically investigated, which led to the isolation of fifty compounds including eight new compounds (M1-M8). Their structures with absolute configuration were elucidated by detailed analyses of spectroscopic data (IR, MS, NMR, UV), CD, and single-crystal X-ray diffraction. The new compounds included six new oxazole-containing diterpenoids (M1-M6) and two norditerpenoids (M7-M8). Especially, the single-crystal X-ray diffraction experiment was the first time for confirming the oxazole ring and the absolute configuration of this type diterpenoids. The plausible biosynthetic pathway of aromatic oxazole-containing diterpenoids was also proposed.The isolated compounds were evaluated for their cytotoxicities, antiviral activities, neuroprotective activities. Compounds M19-M21, M31-M33showed selected and significant activity against several cancer cell lines. Several compounds displayed potent activity against HIV-1replication. It’s interesting that the values of inhibitory activities of these compounds are not concentration-dependent, the values of inhibitory activities of2compounds in0.1μM were in the range of40-50%. Compounds M19-M23, M31, M33inhibited lipopolysaccharide (LPS)-induced pro-inflammatory factors in BV2-macrophages with the inhibition ratio of100%at10μM. The results imply that plant cell cultures can not only produce known bioactive compounds, but also yield new bioactive metabolites, which offer alternative and effective sources of lead compounds.