title: Isolation and characterization of bioactive metabolites from Xylaria psidii, an endophytic fungus of the medicinal plant Aegle marmelos and their role in mitochondrial dependent apoptosis against pancreatic cancer cells.
abstract: BACKGROUND
  The genus Xylaria has been reported as a rich source of biologically active secondary metabolites. In the present study, an endophytic fungus Xylaria psidii has been isolated from the leaf sample of Aegle marmelos (L.) Corr., characterized on the basis of its morphological features and sequence data for the ITS region (KU291350) of the nuclear ribosomal DNA. Biological screening of ethyl acetate extract of Xylaria psidii displayed a potential therapeutic effect on pancreatic cancer cells.
  
  
  OBJECTIVE
  This study was designed systematically to explore Xylaria psidii, an endophytic fungus for the identification of biologically active secondary metabolites against pancreatic cancer cells.
  
  
  METHODS
  While exploring the bioactive secondary metabolites, a sensitive and reliable LC-MS based dereplication approach was applied to identify four compounds A-D from fungal extract. Further bioactivity guided isolation of fungal extract yielded two major metabolites 1 and 2. The structures of 1 and 2 have been determined by detailed spectroscopic analysis including MS, NMR, IR and UV data and similarity with published data. Xylarione A (1) is new whereas (-) 5-methylmellein (2) is reported for the first time from X. psidii. Both the isolated compounds were screened for their effect on the viability and proliferation against a panel of cancer cell lines (MCF-7, MIA-Pa-Ca-2, NCI-H226, HepG2 and DU145) of different tissue origin.
  
  
  RESULTS
  Compounds 1 and 2 exhibited cytotoxicity against pancreatic cancer (MIA-Pa-Ca-2) cells with IC50values of 16.0 and 19.0 µm, respectively. The cell cycle distribution in MIA-Pa-Ca-2 cells, confirmed a cell cycle arrest at the sub-G1 phase. Cell death induced by 1 and 2 displayed features characteristic of apoptosis. Flow cytometry based analysis of 1 and 2 using Rhodamine-123 displayed substantial loss of mitochondrial membrane potential in a concentration dependent manner by both the compounds.
  
  
  CONCLUSIONS
  Results conclude that the isolated compounds 1 and 2 are responsible for the activity shown by crude ethyl acetate extract and may act as potential leads for medicinal chemists for designing more potent analogs.
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India.
authors.forename: Divya
authors.initials: D
authors.lastname: Arora
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India.
authors.forename: Nisha
authors.initials: N
authors.lastname: Sharma
authors.affiliation: Natural Product Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India.
authors.forename: Venugopal
authors.initials: V
authors.lastname: Singamaneni
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India.
authors.forename: Vishal
authors.initials: V
authors.lastname: Sharma
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kushwaha
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India.
authors.forename: Vidushi
authors.initials: V
authors.lastname: Abrol
authors.affiliation: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India.
authors.forename: Santosh
authors.initials: S
authors.lastname: Guru
authors.affiliation: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India.
authors.forename: Sonia
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India.
authors.forename: Ajai Prakash
authors.initials: AP
authors.lastname: Gupta
authors.affiliation: Cancer Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India. Electronic address: sbhushan@iiim.ac.in.
authors.forename: Shashi
authors.initials: S
authors.lastname: Bhushan
authors.affiliation: Quality Control & Quality Assurance Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India. Electronic address: sundeepjaglan@iiim.ac.in.
authors.forename: Sundeep
authors.initials: S
authors.lastname: Jaglan
authors.affiliation: Natural Product Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu 180001, India; Academy of Scientific & Innovative Research (AcSIR), CSIR, New Delhi, 110025, India. Electronic address: guptap@iiim.ac.in.
authors.forename: Prasoon
authors.initials: P
authors.lastname: Gupta
mesh_terms: D000085:Acetates; D045471:Aegle / Q000737:chemistry* / Q000382:microbiology*; D000903:Antibiotics, Antineoplastic / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001203:Ascomycota / Q000737:chemistry*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D060026:Endophytes / Q000737:chemistry*; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects*; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D012997:Solvents
chemical_list: D000085:Acetates; D000903:Antibiotics, Antineoplastic; D012997:Solvents; C007650:ethyl acetate
keywords: Apoptosis; Cytotoxicity; Endophytic fungi; MIA-Pa-Ca-2 cells; Xylaria psidii