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Piperazine ring toxicity in three novel anti-breast cancer drugs: an in silico and in vitro metabolic bioactivation approach using olaparib as a case study

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Abstract

The metabolic activation of small-molecule drugs into electrophilic reactive metabolites is widely recognized as an indicator of idiosyncratic adverse drug reactions (IADRs). Three novel anti-breast cancer drugs containing piperazine rings, ribociclib (Kisqali®, RCB), abemaciclib (Verzenio®, ABC), and olaparib (Lynparza®, OLP), were selected to study the effect of different chemical environment on the piperazine ring activation using in silico and in vitro metabolic experiments. ABC and RCB were previously studied and we noticed the piperazine ring in ABC could be strongly bioactivated generating more reactive intermediates than piperazine ring in RCB. OLP was further used as a case study to show the power of in silico software to predict the piperazine ring activation that was approved using in vitro experiments. Initially, predictions of susceptible sites in the metabolism and reactivity pathways were performed using the StarDrop P450 model and XenoSite reactivity tool, respectively. Later, in vitro OLP metabolites were characterized based on rat liver microsomes (RLMs) using KCN (trapping agent) using LC–MS/MS. The main goal of the current study was to answer the question of whether the presence of a piperazine ring in the chemical structure should be always considered a structural alert. Piperazine ring in RBC and ABC was bioactivated through a metabolic sequence that involves the hydroxylation of α-carbon to the tertiary nitrogen atoms of the piperazine ring. In the case of OLP, no cyano adduct was formed due to the presence of two carbonyl groups attached to the two nitrogen atoms of the piperazine ring (neutral amide groups). From the results, piperazine ring in certain cases should not be considered as a structural alert as in the case of OLP due to the presence of two electron withdrawing group that stops the proposed toxicity. Also blocking the bioactive center (α-carbon) using steric hindrances such as methyl group, also the isosteric replacement of α-carbon hydrogen with fluoro atom can aid in reducing the toxic side effects of ABC and RCB. These experiments were done in vitro through incubation with RLMs in the presence of KCN. Also, the results are supported by data generated from in silico software. In the future, drug discovery studies using this concept could be undertaken, allowing for the development of new drugs with reasonable safety profiles. Overall, in vitro RLMs incubations and in silico experiments were able to predict successfully that the piperazine ring should not always be considered a structural alert.

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Acknowledgements

The authors extend their appreciation to the Deputyship for Research & Innovation, “Ministry of Education” in Saudi Arabia for funding this research work through project no. (IFKSURG-2-708).

Funding

This study was funded by the Deputyship for Research & Innovation, “Ministry of Education” in Saudi Arabia through project no. (IFKSURG-2-708).

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Authors and Affiliations

  1. Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh, 11451, Saudi Arabia

    Thamer A. Alsubi, Mohamed W. Attwa, Hany W. Darwish, Hatem A. Abuelizz & Adnan A. Kadi

  2. Students’ University Hospital, Mansoura University, Mansoura, 35516, Egypt

    Mohamed W. Attwa

  3. Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini St., Cairo, 11562, Egypt

    Hany W. Darwish

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  1. Thamer A. Alsubi
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  2. Mohamed W. Attwa
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  3. Hany W. Darwish
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  4. Hatem A. Abuelizz
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  5. Adnan A. Kadi
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Contributions

AK, HD, TS, and HA contributed to the study design. TS and MA performed the data analysis and experimental work and aided in drafting the manuscript. AK, HA, and HD directed the experimental laboratory work. MA is the corresponding author of this paper. The final draft of the manuscript was revised by all the authors. All authors reviewed, read, and approved the manuscript. All experimental data were generated in-house and no paper mill was used.

Corresponding author

Correspondence to Mohamed W. Attwa.

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Ethical approval

The male Sprague Dawley rats were maintained at King Saud University following the animal care center instructions, which have been approved by the Local Animal Care and Use Committee of the university. The animal experiment procedure used in the current research was validated and approved by King Saud University’s Ethics Review Committee (No.: KSU-SE-19–76).

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The authors declare no competing interests.

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Alsubi, T.A., Attwa, M.W., Darwish, H.W. et al. Piperazine ring toxicity in three novel anti-breast cancer drugs: an in silico and in vitro metabolic bioactivation approach using olaparib as a case study. Naunyn-Schmiedeberg's Arch Pharmacol 396, 1435–1450 (2023). https://doi.org/10.1007/s00210-023-02413-9

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