Pyrophen Produced by Endophytic Fungi Aspergillus sp Isolated from Piper crocatum Ruiz & Pav Exhibits Cytotoxic Activity and Induces S Phase Arrest in T47D Breast Cancer Cells

  • Astuti, Puji (Pharmaceutical Biology Department, Faculty of Pharmacy, Universitas Gadjah Mada) ;
  • Erden, Willy (Chemistry Department, Faculty of Mathematics and Natural Sciences, Semarang State University) ;
  • Wahyono, Wahyono (Pharmaceutical Biology Department, Faculty of Pharmacy, Universitas Gadjah Mada) ;
  • Wahyuono, Subagus (Pharmaceutical Biology Department, Faculty of Pharmacy, Universitas Gadjah Mada) ;
  • Hertiani, Triana (Pharmaceutical Biology Department, Faculty of Pharmacy, Universitas Gadjah Mada)
  • Published : 2016.03.07


Ethyl acetate extracts obtained from culture of endophytic fungi Aspergillus sp isolated from Piper crocatum Ruiz & Pav, have been shown to possess cytotoxic activity against T47D breast cancer cells. Investigations were here conducted to determine bioactive compounds responsible for the activity. Bioassay guided fractionation was employed to obtain active compounds. Structure elucidation was performed based on analysis of LC-MS, $^1H$-NMR, $^{13}C$-NMR, COSY, DEPT, HMQC, HMBC data. Cytotoxity assays were conducted in 96 well plates against T47D and Vero cell lines. Bioassay guided isolation and chemical investigation led to the isolation of pyrophen, a 4-methoxy-6-(1'-acetamido-2'-phenylethyl)-2H-pyran-2-one. Further analysis of its activity against T47D and Vero cells showed an ability to inhibit the growth of T47D cells with IC50 values of $9.2{\mu}g/mL$ but less cytotoxicity to Vero cells with an $IC_{50}$ of $109{\mu}g/mL$. This compound at a concentration of 400 ng/mL induced S-phase arrest in T47D cells.


Aspergillus sp;cytotoxic;T47D;pyrophen;S;phase arrest


Supported by : Universitas Gadjah Mada


  1. Astuti P, Wahyono, Nababan OA (2014). Antimicrobial and cytotoxic activities of endophytic fungi isolated from Piper crocatum Ruiz & Pav. Asian Pac J Trop Biomed, 4, 643-7.
  2. Barnes CL, Steiner JR, Torres E, et al (1990). Structure and absolute configuration of pyrophen, a novel pryrone derivative of L-phenylalanine from Aspergillus niger. Int. J Peptide Protein Res, 36, 292-6.
  3. Chen H, Daletos G, Okoye F, et al (2015). A new cytotoxic cytochalasin from the endophytic fungus Trichoderma harzianum. Nat Prod Commun, 10, 585-7.
  4. Cui JL, Guo SX, Xiao PG (2011). Antitumor and antimicrobial activities of endophytic fungi from medicinal parts of Aquilaria sinensis, J Zhejiang Univ-Sci B (Biomed & Biotechnol), 12, 385-92.
  5. do Nascimento AM, Soares MG, da Silva Torchelsen FK, et al (2015). Antileishmanial activity of compounds produced by endophytic fungi derived from medicinal plant Vernonia polyanthes and their potential as source of bioactive substances. World J Microbiol Biotechnol, 31, 1793-800.
  6. IARC (2012) Globocan 2012 : estimated cancer insidence, mortality and prevalence worldwide in 2012. (available online 16 October 2015).
  7. IARC. (2008).World cancer report 2008. Lyon, International Agency for Research on Cancer. (available online 16 October 2015.)
  8. Kaufmann WK, Kaufman DG (1993). Cell cycle control, DNA repair and initiation of carcinogenesis. FASEB J, 7, 1188-91.
  9. Richardson SN, Nsiama TK, Walker AK, et al (2015). Antimicrobial dihydrobenzofurans and xanthenes from a foliar endophyte of Pinus strobus. Phytochemistry, 117, 436-43.
  10. Santiago C, Fitchett C, Munro MH, et al (2012). Cytotoxic and Antifungal Activities of 5-Hydroxyramulosin, a Compound Produced by an Endophytic Fungus Isolated from Cinnamomum mollisimum. Evid Based Complement Alternat Med, 689310.
  11. Shaaban M, Shaaban KA, Abdel-Aziz MS (2012). Seven naphtho-${\gamma}$-pyrones from the marine-derived fungus Alternaria alternata: structure elucidation and biological properties. Organic and Medicinal Chemistry Letters, 2, 6.
  12. Varoglu M, Crews P (2000). Biosynthetically diverse compounds from a saltwater culture of sponge-derived Aspergillus niger. J Natural Products, 63, 41-3.
  13. Wang FQ, Tong QY, Ma HR, et al (2015). Indole diketopiperazines from endophytic Chaetomium sp 88194 induce breast cancer cell apoptotic death. Sci Rep, 5, 9294.
  14. WHO (2013). Cancer. (available online 16 October 2015).
  15. Zhang Y, Li XM, Feng Y, et al (2010). Phenethyl-${\alpha}$-pyrone derivatives and cyclodipeptides from a marine algous endophytic fungus Aspergillus niger EN-13, Natural Product Research: Formerly Natural Product Letters, 24, 1036-43.
  16. Zhao K, Ping W, Li Q, et al (2009). Aspergillus niger var. taxi, a new species variant of taxol-producing fungus isolated from Taxus cuspidate in China. J Appl Microbiol, 107, 1202-7.
  17. Zheng CJ, Xu LL, Li YY, et al (2013). Cytotoxic metabolites from the cultures of endophytic fungi from Panax ginseng. Appl Microbiol Biotechnol, 97, 7617-25.
  18. Zhu F, Chen G, Wu J, et al (2013). Structure revision and cytotoxic activity of marinamide and its methyl ester, novel alkaloids produced by co-cultures of two marine-derived mangrove endophytic fungi. Nat Prod Res, 27, 1960-4.

Cited by

  1. pp.1478-6427, 2019,