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Induction of endoplasmic reticulum stress by aminosteroid derivative RM-581 leads to tumor regression in PANC-1 xenograft model

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Summary

The high fatality and morbidity of pancreatic cancer have remained almost unchanged over the last decades and new clinical therapeutic tools are urgently needed. We determined the cytotoxic activity of aminosteroid derivatives RM-133 (androstane) and RM-581 (estrane) in three human pancreatic cancer cell lines (BxPC3, Hs766T and PANC-1). In PANC-1, a similar level of antiproliferative activity was observed for RM-581 and RM-133 (IC50 = 3.9 and 4.3 μM, respectively), but RM-581 provided a higher selectivity index (SI = 12.8) for cancer cells over normal pancreatic cells than RM-133 (SI = 2.8). We also confirmed that RM-581 induces the same ER stress-apoptosis markers (BIP, CHOP and HERP) than RM-133 in PANC-1 cells, pointing out to a similar mechanism of action. Finally, these relevant in vitro results have been successfully translated in vivo by testing RM-581 using different doses (10–60 mg/kg/day) and modes of administration in PANC-1 xenograft models, which have led to tumor regression without any sign of toxicity in mice (animal weight, behavior and histology). Interestingly, RM-581 fully reduced the pancreatic tumor growth when administered orally in mice.

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Abbreviations

AM:

Aminosteroid derivative

AUC:

Area under the curve

CCAC:

Canadian Council on Animal Care

CI:

Combination index

Cmax :

Maximal concentration

DMSO:

Dimethyl sulfoxide

EDTA:

Ethylenediamine tetraacetic acid

ER:

Endoplasmic reticulum

HE:

Hematoxylin and eosin

IC50 :

Concentration inhibiting 50% of cell growth

ip :

Intraperitoneally

LC/MS/MS:

Liquid chromatography/mass spectrometry/mass spectrometry

MT:

Masson’s trichrome

MTS:

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfophen-yl)-2H-tetrazolium

PAS:

Periodic acid Schiff

PAS-D:

Periodic acid Schiff with diastase

PBS:

Phosphate buffer solution

PG:

Propylene glycol

PK:

Pharmacokinetic

po :

Orally

sc :

Subcutaneously

SI:

Selectivity index

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Acknowledgments

We thank Sonia Francoeur (animal facilities) and Patrick Caron (LC-MS/MS analyses) for their technical assistance. We would also like to thank Micheline Harvey for careful reading of the manuscript, and The National Institutes of Health (Bethesda, MD, USA) for providing the antineoplastic drugs.

Funding

This work was supported by the Canadian Institutes of Health Research (CIHR) from the Targeting High Fatality Cancers – Innovation Grant program and the Proof of Principle-Phase I program.

Author information

Author notes

  1. Martin Perreault and Jenny Roy contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Medicinal Chemistry, Endocrinology and Nephrology Unit, CHU de Québec - Research Center (CHUL, T4-42), 2705 Laurier Boulevard, Québec, QC, GIV 4G2, Canada

    Martin Perreault, René Maltais, Jenny Roy & Donald Poirier

  2. Department of Anatomo-Pathology, CHU de Québec - Université Laval, Québec, QC, GIV 4G2, Canada

    Sylvain Picard & Ion Popa

  3. Faculty of Pharmacy, Université Laval, Québec, QC, GIV OA6, Canada

    Nicolas Bertrand

  4. Endocrinology and Nephrology Unit, CHU de Québec-Research Center (CHUL, T4-13), Québec, QC, GIV 4G2, Canada

    Nicolas Bertrand

  5. Department of Molecular Medicine, Faculty of Medicine, Université Laval, Québec, QC, GIV OA6, Canada

    Donald Poirier

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  1. Martin Perreault
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Correspondence to Donald Poirier.

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Conflict of interest

MP, RM, JR and DP have ownership interests on patent applications and patents related to these families of aminosteroid derivatives. SP, IP and NB declare no conflict of interest.

Electronic supplementary material

Additional information related to: 1) Plasmatic concentration of RM-581; 2) Quantification of RM-581 in plasma; 3) PANC-1 xenograft protocol; 4) Supplementary Table 1: qPCR primers details; 5) Supplementary Table 2: RM-581 is cytotoxic for pancreatic cancer cells and 6) Supplementary Fig. 1: Animal and pancreas weights of mice treated with RM-581.

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Perreault, M., Maltais, R., Roy, J. et al. Induction of endoplasmic reticulum stress by aminosteroid derivative RM-581 leads to tumor regression in PANC-1 xenograft model. Invest New Drugs 37, 431–440 (2019). https://doi.org/10.1007/s10637-018-0643-4

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