Sensitivity of different resistant tumour cell lines to the two novel compounds (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate and (1E,3E)-1,4-bis(2-naphthyl)-2,3-dinitro-1,3-butadiene

doi:10.1016/j.ejphar.2008.04.020

European Journal of Pharmacology

Volume 588, Issue 1, 24 June 2008, Pages 47-51

https://doi.org/10.1016/j.ejphar.2008.04.020 Get rights and content

Abstract

The inhibition of cell proliferation by methyl (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate (1-Naph-NMCB) and (1E,3E)-1,4-bis(2-naphthyl)-2,3-dinitro-1,3-butadiene (2-Naph-DNB) has been studied in vitro against four cell lines selected for their resistance to doxorubicin, cisplatin, taxol and 5-fluorouracil. In previous experiments both compounds showed good in vitro antiproliferative, cytotoxic and pro-apoptotic activities against cell lines of different histologic origin.

The results of the experiments presented here suggest that 1-Naph-NMCB is able to overcome all of the different mechanisms of resistance showed by the resistant cell lines used for our experiments. On the contrary, when we used the taxol-resistant A549-T12 cell line, characterized by a mechanism of resistance due to a mutation of the target site of taxol on microtubules, it displayed a partial but significant cross-resistance to 2-Naph-DNB. Although the actual mechanism of this cross-resistance has not yet been definitively elucidated, our results from immunostaining of microtubules suggest that it may be linked to the presence of a shared target site for taxol and 2-Naph-DNB on microtubules.

Introduction

In an attempt to generate new antitumour compounds with good properties in terms of antiproliferative and pro-apoptotic activity as well as of low toxicity in vivo and ability to overcome the most frequent mechanisms of resistance responsible for the failure of a high number of chemotherapeutic cancer treatments, we synthesized (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate (1-Naph-NMCB, Petrillo et al., 2008) and (1E,3E)-1,4-bis(2-naphthyl)-2,3-dinitro-1,3-butadiene (2-Naph-DNB, Viale et al., 2007). The first compound is basically the result of the application of the “molecular simplification strategy” (Manetti et al., 2000, Crisòstomo et al., 2006) on the lead compound (1E,3E)-1,4-bis(1-naphthyl)-2,3-dinitro-1,3-butadiene (1-Naph-DNB, Viale et al., 2004, Novi et al., 2004, Dell'Erba et al., 2005), whereas the second molecule is a structural isomer of the latter, characterized by a different spatial arrangement (Fig. 1).

When tested in vitro for its inhibition of cell proliferation and apoptotic activity 1-Naph-NMCB showed a significant activity at micromolar concentrations, in particular against the MDA-MB-231 breast cancer cell line, and with a significant general improvement compared to 1-Naph-DNB (Petrillo et al., 2008). In particular, for apoptosis, 1-Naph-NMCB showed an activity that was in some cases better than that observed for 1-Naph-DNB, as evaluated by the morphological analysis of nuclear segmentation, the staining with Annexin V and the analysis in western blot of p53 oncosuppressor protein (Petrillo et al., 2008). Moreover, both the analysis of formation of interstrand cross-links and the analysis of the inhibition of restriction enzyme cutting activity in λ phage DNA treated with the compound (unpublished data) demonstrate the binding of 1-Naph-NMCB to DNA, although lower than that observed for the parent compound 1-Naph-DNB (Petrillo et al., 2008).

In vivo studies allowed the toxicological (determination of lethal and maximal tolerated doses) and pharmacological evaluation of 1-Naph-NMCB. Our findings showed that our compound was characterized by negligible histological toxic effects and an antitumour activity which was in some cases even better than that showed by 1-Naph-DNB, and was characterized by differences in tumour selectivity (Petrillo et al., 2007).

2-Naph-DNB showed significant features in terms of inhibition of cell proliferation, with a better activity than 1-Naph-DNB in MDA-MB-231 cells and HGC-27 human gastric cell line, and induction of apoptosis which was linked to the activation of p53 protein and the formation of interstrand cross-links to DNA (Viale et al., 2007). The latter observation was also confirmed by experiments using treated λ phage DNA and restriction enzymes (unpublished data). As for 1-Naph-NMCB, also 2-Naph-DNB was characterized in vivo by low toxic effect at histological level.

All these features prompted us to verify the ability of our molecules to overcome the different mechanisms of resistance showed by some cell lines selected in vitro for their resistance to important anticancer drugs such as doxorubicin, taxol, cisplatin and 5-fluorouracil, also in order to gain further insights into their mechanisms of action.

Section snippets

Chemicals

The two naphthylnitrobutadienes 2-Naph-DNB and 1-Naph-NMCB were synthesized from 3,4-dinitrothiophene (3,4-DNT) (Viale et al., 2007) and methyl 4-nitrothiophene-2-carboxylate (2-COOMe-4-NT) (Petrillo et al., 2007) as already described (Scheme 1).

While 1-Naph-NMCB differs from the lead compound 1-Naph-DNB (Petrillo et al., 2008) since it contains only one of the two original naphthylnitroethenyl moieties, the isomeric 2-Naph-DNB simply differs from 1-Naph-DNB for a different spatial arrangement

Inhibition of cell proliferation and morphology of microtubules

In human A2780 cells, sensitive (doubling time 24 h) and resistant to doxorubicin (doubling time 27 h), both 1-Naph-NMCB and 2-Naph-DNB were active at the micromolar concentration level. Both molecules were less active than doxorubicin in sensitive A2780 cells, on the other hand their IC₅₀s for resistant and sensitive cells were nearly similar. On the basis of the results showed in Table 1, the resistance indexes for doxorubicin, 1-Naph-NMCB and 2-Naph-DNB were 63.0, 1.8 and 1.2, respectively.

Discussion

As reported in a previous paper (Novi et al., 2004) the mechanisms of resistance of the cell lines used in our experiments are quite well characterized. All these cells were selected in vitro by prolonged exposure to the different selective anticancer drugs.

Briefly, the main mechanism of resistance of L1210/DDP cells consists of a decreased accumulation of cisplatin and a reduced DNA platination (Richon et al., 1987). The resistance of HCT-8/FU7dR cell lines was linked to the shortage of

Acknowledgements

This research was supported by funds from M.I.U.R. (FIRB 2001) and from the Universities of Genova and Bologna.

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Short communicationSensitivity of different resistant tumour cell lines to the two novel compounds (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate and (1E,3E)-1,4-bis(2-naphthyl)-2,3-dinitro-1,3-butadiene

Abstract

Introduction

Section snippets

Chemicals

Inhibition of cell proliferation and morphology of microtubules

Discussion

Acknowledgements

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J. Biol. Chem.

Biorg. Med. Chem.

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Relationship between cytotoxicity, drug accumulation, DNA damage and repair of human ovarian cancer cells treated with doxorubicin: modulation by the tiapamil analog RO11-2933

Cancer Chemother. Pharmacol.

Novel mechanism(s) of resistance to 5-fluorouracil in human colon cancer (HCT-8) sublines following exposure to two different clinically relevant dose schedules

Cancer Res.

Molecular simplification in bioactive molecules: formal synthesis of (+)-muconin

J. Org. Chem.

Short communication
Sensitivity of different resistant tumour cell lines to the two novel compounds (2Z,4E)-2-methylsulfanyl-5-(1-naphthyl)-4-nitro-2,4-pentadienoate and (1E,3E)-1,4-bis(2-naphthyl)-2,3-dinitro-1,3-butadiene