Isolation, structural modification of macrophin from endophytic fungus Phoma macrostoma and their cytotoxic potential

  • Yedukondalu Nalli
  • Palak Arora
  • Sameer Khan
  • Fayaz Malik
  • Syed Riyaz-Ul-Hassan
  • Vivek Gupta
  • Asif AliEmail author
Original Research


Four known metabolites, macrophin (1), rosellisin (2), 2-(2-hydroxy-5-6-methoxy-3-methylene-1,4-benzodioxin-2(3H)-one (3), and methoxyphenoxyacrylic acid (4) were isolated, for the first time, from an endophytic fungus, Phoma macrostoma inhabiting the inner tissue of medicinal plant Glycyrrhiza glabra Linn. Their structures were characterized by comparison of their NMR data with literature data, and X-ray diffraction data of macrophin (1) as described herein for the first time. The compounds (14) were evaluated for their growth-inhibitory activities against a panel of cancer cell lines. Of these substances, macrophin (1) showed prominent cytotoxic activity against the MDA-MB-231, T47D, MCF-7, and MIAPaCa-2 cancer-cell lines with IC50 values of 14.8, 8.12, 13.0, and 0.9 μM, respectively. This biological significance has prompted us to generate a series of five acylated analogs (1a–1e) of 1 to conduct structure–activity relationship (SAR) study. The result illustrates all the analogues (1a–1e) were more potent than 1 against MDMB-231 and MIAPaCa-2 cancer cell lines. The compound 1d, an n-butanoyl substituent, has been proven to be three-fold more potent than 1 with IC50 values of 0.3 and 3.7 µM against MIAPaCa-2 and MDMB-231 cell lines, respectively.


Phoma macrostoma Isolation Macrophine Semi-synthesis Cytotoxic activities 



PA is supported by the Department of Science and Technology, New Delhi, India through INSPIRE Senior Research Fellowship. PA and SK acknowledge AcSIR for their enrollment in the Ph.D. program. The authors would like to acknowledge the Ministry of AYUSH (New Delhi) for providing financial support for the work of YN (Z.28015/229/2015-HCP EMR). The article bears the institutional manuscript no. IIIM/2271/2018.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

44_2018_2281_MOESM1_ESM.docx (6.9 mb)
Supporting information


  1. Almasan A, Yin Y, Kelly RE, Lee EY, Bradley A, Li W, Bertino JR, Wahl GM (1995) Deficiency of retinoblastoma protein leads to inappropriate S-phase entry, activation of E2F-responsive genes, and apoptosis. Proc Natl Acad Sci USA 92:5436–5440CrossRefGoogle Scholar
  2. Alurappa RCS, Narayanaswamy R, Sinniah UR, Mohanty SK, Swamy MK (2018) Endophytic fungi and bioactive metabolites production: an update. In: Patra J, Das G, Shin HS(eds) Microbial Biotechnology. Springer, SingaporeGoogle Scholar
  3. Arora P, Wani ZA, Nalli Y, Ali A, Riyaz-Ul-Hassan S (2016) Antimicrobial potential of Thiodiketopiperazine derivatives produced by Phoma sp., an endophyte of Glycyrrhiza glabra Linn. Microb Ecol 72:802–812CrossRefGoogle Scholar
  4. Assante G, Camarda L, Merlini L, Nasini G (1981) Secondary metabolites from Mycosphaerella ligulicola. Phytochemistry 20:1955–1957CrossRefGoogle Scholar
  5. Aveskamp MM, de Gruyter J, Woudenberg JHC, Verkley GJM, Crous PW (2010) Highlights of the Didymellaceae: a polyphasic approach to characterise Phoma and related pleosporalean genera. Stud Mycol 65:1–60CrossRefGoogle Scholar
  6. Bennett A, Ponder M, Garcia-Diaz J (2018) Phoma infections: classification, potential food sources, and their clinical impact. Microorganisms 6:58CrossRefGoogle Scholar
  7. Butler MS (2004) The role of natural product chemistry in drug discovery. J Nat Prod 67:2141–2153CrossRefGoogle Scholar
  8. Butler MS (2005) Natural products to drugs: natural product derived compounds in clinical trials. Nat Prod Rep 22:162–195CrossRefGoogle Scholar
  9. Crombie L, Crombie WML, Jamieson SV, Palmer CJ (1988) Acid catalyzed terpenylations of olivetol in the synthesis of cannabinoids. J Chem Soc Perkin Trans 1:1243–1250CrossRefGoogle Scholar
  10. Estrov Z, Shishodia S, Faderl S, Harris D, Van Q, Kantarjian HM, Talpaz M, Aggarwal BB (2003) Resveratrol blocks interleukin-1beta-induced activation of the nuclear transcription factor NF-kappaB, inhibits proliferation, causes S-phase arrest, and induces apoptosis of acute myeloid leukemia cells. Blood 102:987–995CrossRefGoogle Scholar
  11. Fulda S, Debatin KM (2006) Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy. Oncogene 25:4798CrossRefGoogle Scholar
  12. Gerl R, Vaux DL (2005) Apoptosis in the development and treatment of cancer. Carcinogenesis 26:263–270CrossRefGoogle Scholar
  13. Graupner PR, Carr A, Clancy E, Gilbert J, Bailey KL, Derby JA, Gerwick BC (2003) The macrocidins: novel cyclic tetramic acids with herbicidal activity produced by Phoma macrostoma. J Nat Prod 66:1558–1561CrossRefGoogle Scholar
  14. Jia M, Chen L, Xin H-L, Zheng C-J, Rahman K, Han T, Qin L-P (2016) A friendly relationship between endophytic fungi and medicinal plants: a systematic review. Front Microbiol 7:906CrossRefGoogle Scholar
  15. Kumar S, Guru SK, Venkateswarlu V, Malik F, Vishwakarma RA, Sawant SD, Bhushan S (2015) A novel quinoline based second-generation mTOR inhibitor that induces apoptosis and disrupts PI3K-Akt-mTOR signaling in human leukemia HL-60 cells. Anti-Cancer Agents Med Chem 15:1297–1304CrossRefGoogle Scholar
  16. Kusari S, Hertweck C, Spiteller M (2012) Chemical ecology of endophytic fungi: origins of secondary metabolites. Chem Biol 19:792–798CrossRefGoogle Scholar
  17. Nair MSR (1976) Biogenesis and revised structure of rosellisin; structure of rosellisin aldehyde. Phytochemistry 15:1090–1091CrossRefGoogle Scholar
  18. Nalli Y, Arora P, Wadhwa B, Malik FA, Vishwakarma RA, Gupta VK, Riyaz-Ul-Hassan S, Ali A (2017) Diapolic acid A-B from an endophytic fungus, Diaporthe terebinthifolii depicting antimicrobial and cytotoxic activity. J Antibiot 70:212–215CrossRefGoogle Scholar
  19. Nalli Y, Mirza DN, Wani ZA, Wadhwa B, Mallik FA, Raina C, chaubey A, Riyaz-Ul-Hassan S, Ali A (2015) Phialomustin A–D, new antimicrobial and cytotoxic metabolites from an endophytic fungus, Phialophora mustea. RSC Adv 5:95307–95312CrossRefGoogle Scholar
  20. Pusztahelyi T, Holb I, Pócsi I (2015) Secondary metabolites in fungus–plant interactions. Front Plant Sci 6:573CrossRefGoogle Scholar
  21. Qadri M, Rajput R, Abdin MZ, Vishwakarma RA, Riyaz-Ul-Hassan S (2014) Diversity, molecular phylogeny, and bioactive potential of fungal endophytes associated with the Himalayan blue pine (Pinus wallichiana). Microb Ecol 67:877–887CrossRefGoogle Scholar
  22. Rubinsztein DC, Gestwicki JE, Murphy LO, Klionsky DJ (2007) Potential therapeutic applications of autophagy. Nat Rev Drug Discov 6:304–312CrossRefGoogle Scholar
  23. Strobel G, Singh SK, Riyaz-Ul-Hassan S, Mitchell AM, Geary B, Sears J (2011) An endophytic/pathogenic Phoma sp. from creosote bush producing biologically active volatile compounds having fuel potential. FEMS Microbiol Lett 320:87–94CrossRefGoogle Scholar
  24. Wang X, Zhou H, Chen H, Jing X, Zheng W, Li R, Sun T, Liu J, Fu J, Huo L, Li Y-z, Shen Y, Ding X, Müller R, Bian X, Zhang Y (2018) Discovery of recombinases enables genome mining of cryptic biosynthetic gene clusters in Burkholderiales species. Proc Natl Acad Sci USA 115:E4255–E4263PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yedukondalu Nalli
    • 1
  • Palak Arora
    • 2
    • 3
  • Sameer Khan
    • 3
    • 4
  • Fayaz Malik
    • 3
    • 4
  • Syed Riyaz-Ul-Hassan
    • 2
    • 3
  • Vivek Gupta
    • 5
  • Asif Ali
    • 1
    • 2
    Email author
  1. 1.Natural Product Chemistry DivisionCSIR-Indian Institute of Integrative MedicineJammuIndia
  2. 2.Microbial Biotechnology DivisionCSIR-Indian Institute of Integrative MedicineJammuIndia
  3. 3.Academy of Scientific and Innovative ResearchCSIR-Indian Institute of Integrative MedicineJammuIndia
  4. 4.Pharmacology DivisionCSIR-Indian Institute of Integrative MedicineJammuIndia
  5. 5.Post-Graduate Department of Physics & ElectronicsUniversity of JammuJammuIndia

Personalised recommendations