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Novel Very Long-Chain α-Methoxylated Δ5,9 Fatty Acids from the Sponge Asteropus niger Are Effective Inhibitors of Topoisomerases IB

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Lipids

Abstract

The novel fatty acids (2R,5Z,9Z)-2-methoxy-25-methyl-5,9-hexacosadienoic acid (1a) and (2R,5Z,9Z)-2-methoxy-24-methyl-5,9-hexacosadienoic acid (1b) were isolated in 80 % purity from the Caribbean sponge Asteropus niger by chloroform/methanol extraction followed by solvent partitioning and silica gel column chromatography. The compounds were characterized by utilizing a combination of gas chromatography-mass spectrometry, nuclear magnetic resonance, and circular dichroism. Acids 1a and 1b were not detected in the phospholipids (PtdCho and PtdIns) of the sponge, but rather as free FA and possibly in glycosylceramides. The mixtures of 1a and 1b displayed cytotoxicity towards THP-1 and HepG2 cells with EC50’s between 41 and 35 μg/mL. Apoptosis was not the preferred mode of cell death induced by 1a–1b in the THP-1 cells. This implies other types of cytotoxicity mechanisms, such as membrane disruption and/or the inhibition (EC50 = 1.8 μg/mL) of the human topoisomerase IB enzyme (hTopIB), with a mechanism of inhibition different from the one displayed by camptothecin (CPT). In a separate experiment, the mixture of 1a and 1b also displayed cytotoxicity towards ex vivo mouse splenocytes infected with Leishmania infantum amastigotes (IC50 = 0.17 mg/mL) and free living promastigotes (IC50 = 0.34 mg/mL). It was also found that the FA were inhibitory of the Leishmania topoisomerase IB (LTopIB) with an EC50 = 5.1 μg/mL. Taken together, 1a and 1b represent a new class of FA with potential as TopIB inhibitors that preferentially inhibit hTopIB over LTopIB.

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Abbreviations

FA:

Fatty acids

CD:

Circular dichroism

CPT:

Camptothecin

ECL:

Equivalent chain length

EC50 :

Half maximal effective concentration

ED50 :

Median effective dose

GC–MS:

Gas chromatography-mass spectrometry

HepG2:

Human hepatocellular liver carcinoma cell line

hTopIB:

Human topoisomerase IB

IC50 :

Half maximal inhibitory concentration

LTopIB:

Leishmania topoisomerase IB

PtdCho:

Phosphatidylcholine

PtdIns:

Phosphatidylinositol

PtdSer:

Phosphatidylserine

iRFP:

Infrared fluorescent protein

THP-1:

Human monocytic leukemia cell line

UPLC-MS:

Ultra high performance liquid chromatography—mass spectrometry

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Acknowledgments

Part of the work described herein was initially supported by Award Number SC1GM084708 from the National Institutes of General Medical Sciences (NIGMS) of the NIH. N. Montano acknowledges the support of the UPR RISE program (Grant No. 2R25GM061151-13) for a graduate fellowship. We thank NIH for the COBRE Mass Spec Core Facility Grant 5P30GM103329-02. We also acknowledge financial support from the Ministerio de Economía y Competitividad (CYTED 214RT0482), Instituto de Salud Carlos III (Feder PI12/00104) and Junta de Castilla y León (grants Gr238 and LE182U13).The technical assistance of J. Vicente in the collection and classification of A. niger is appreciated. We also acknowledge the technical assistance of C. Adorno (UPR-Río Piedras) in the initial isolation work.

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Authors and Affiliations

  1. Department of Chemistry, University of Puerto Rico, Rio Piedras Campus, PO Box 23346, San Juan, PR, 00931-3346, USA

    Néstor M. Carballeira, Nashbly Montano, Luis A. Amador & Abimael D. Rodríguez

  2. Department of Basic Sciences, University of North Dakota, Grand Forks, ND, 58202-9037, USA

    Mikhail Y. Golovko & Svetlana A. Golovko

  3. Department of Biomedical Sciences, University of León, Campus de Vegazana s/n, 24071, León, Spain

    Rosa M. Reguera, Raquel Álvarez-Velilla & Rafael Balaña-Fouce

Authors
  1. Néstor M. Carballeira
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  2. Nashbly Montano
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  3. Luis A. Amador
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  4. Abimael D. Rodríguez
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  5. Mikhail Y. Golovko
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  6. Svetlana A. Golovko
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  7. Rosa M. Reguera
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  8. Raquel Álvarez-Velilla
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  9. Rafael Balaña-Fouce
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Correspondence to Néstor M. Carballeira.

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Carballeira, N.M., Montano, N., Amador, L.A. et al. Novel Very Long-Chain α-Methoxylated Δ5,9 Fatty Acids from the Sponge Asteropus niger Are Effective Inhibitors of Topoisomerases IB. Lipids 51, 245–256 (2016). https://doi.org/10.1007/s11745-015-4114-9

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  • DOI: https://doi.org/10.1007/s11745-015-4114-9

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