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Lactandrate: a D-homo-aza-androsterone alkylator in the treatment of breast cancer

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Summary

The sensitivity of breast neoplasms to hormonal control provides the basis of novel investigational treatments with steroidal alkylators. An androsterone D-lactam steroidal ester, the 3β-hydroxy-13α-amino-13,17-seco- 5α-androstan-17-oic-13,17-lactam, p-bis(2-chloroethyl)amino phenyl acetate (lactandrate) was synthesized and tested for antitumor activity against six human breast cancer cell lines in vitro and against two murine and one xenograft mammary tumors in vivo. A docking study on the binding interactions of lactandrate with the ligand-binding domain (LBD) of estrogen receptor-alpha (ERα) was inquired. In vitro testing of lactandrate cytostatic and cytotoxic activity was performed on T47D, MCF7, MDA-MB-231, BT-549, Hs578T, MDA-MB-435 breast adenocarcinoma human cell lines. In vivo testing was performed on two murine mammary tumors, the MXT tumor and CD8F1 adenocarcinoma, as well as on human mammary carcinoma MX-1 xenograft. Molecular modeling techniques were adopted to predict a possible location and interaction mode of the molecule into LBD. Lactandrate induced significantly high antitumor effect against all tested in vitro and in vivo models. The cell lines with positive ER expression found to be significantly more sensitive to lactandrate. Moreover, lactandrate found to be positioned inside the binding cavity with its steroidal moiety, whilst the alkylating moiety protrudes out of receptor’s pocket. Lactandrate produced important anticancer activity on breast cancer in vitro and in vivo. Some correlation between ER and lactandrate effect was demonstrated. Docking studies provide the basis for the structure-based design of improved steroidal alkylating esters for the treatment of estrogen-related cancers.

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Abbreviations

AF-2:

activation function 2

AWC:

average mice weight change

CR:

complete tumor regression rate

DMSO:

dimethyl sulfoxide

ER:

estrogen receptor

GI50:

50% growth inhibition

hERα:

human estrogen receptor alpha

IC50:

50% in vitro cytotoxicity

i.p.:

intra-peritoneal

LBD:

ligand-binding domain

MTD:

maximum tolerated dose

PKC:

protein kinase C

PR:

partial tumor regression rate

TGI:

total growth inhibition

TFS:

tumor-free survivors

TI:

tumor inhibition

TW:

tumor weight

%T/C:

% median lifespan change of treated animals (T) over the control (C)

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Acknowledgements

This work has been accomplished with the experimental and technical support of the Developmental Therapeutics Program, National Cancer Institute (NCI, USA).

We are grateful to Professor Gabriele Cruciani (Laboratory for Chemometrics, School of Chemistry, University of Perugia, Italy) for the permission to use GRID (v. 22) Linux, free of charge (www.moldiscovery.com).

This study is dedicated to the memory of Professor P. Catsoulacos and Professor G. Wampler.

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Authors and Affiliations

  1. Laboratory of Medicinal Chemistry, Faculty of Pharmacy, University of Patras, Patras, Greece

    Dimitrios T.P. Trafalis & and Charalambos Camoutsis

  2. 1st Department of Medical Oncology, “Metaxa” Cancer Hospital, Piraeus, Greece

    Dimitrios T.P. Trafalis

  3. Symeonidio Research Center, Theagenio Cancer Hospital, Thessaloniki, Greece

    George D. Geromichalos & Athanasios Papageorgiou

  4. Department of Pharmacognosy and Chemistry of Natural Products, School of Pharmacy, University of Athens, Athens, Greece

    Catherine Koukoulitsa

  5. 1st Department of Surgery, Medical School, University of Athens, Athens, Greece

    Panayiotis Karamanakos

  6. 15 Larnakos, 17341 Agios Dimitrios, Athens, Greece

    Dimitrios T.P. Trafalis

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  1. Dimitrios T.P. Trafalis
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  2. George D. Geromichalos
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  3. Catherine Koukoulitsa
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  4. Athanasios Papageorgiou
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  5. Panayiotis Karamanakos
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Correspondence to Dimitrios T.P. Trafalis.

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Trafalis, D.T., Geromichalos, G.D., Koukoulitsa, C. et al. Lactandrate: a D-homo-aza-androsterone alkylator in the treatment of breast cancer. Breast Cancer Res Treat 97, 17–31 (2006). https://doi.org/10.1007/s10549-005-9083-x

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