Synthesis and phospholipidosis effect of a series of cationic amphiphilic compounds: a case study to evaluate in silico and in vitro assays
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In recent years, a large number of in silico and in vitro assays have been developed for safety assessment in early drug discovery. These methods are usually validated using datasets of known drugs with large chemical diversity, while application to homologous series has been rarely explored. Here we report a case study about phospholipidosis (PLD) risk evaluation for a dataset of nine compounds, designed and synthesized to modulate the physico-chemical properties typical of cationic amphiphilic compounds (CADs), representing the main class of PLD inducers. Our aim was to investigate the effect of structure modification on PLD induction according to a number of standard in silico and in vitro methods. As a result, we found that different in silico methods lead to conflicting results when applied to our series of weak PLD inducers, thus the apparently easy-to-use definition of CADs requires special attention. Moreover, when weak inducers are tested in vitro, the revealed PLD effect may vary based on the purity grade of the tested compound and the features of the selected assay. Finally, we have shown that slight modifications on a chemical scaffold can have an impact on the PLD effect. This study also exemplifies that current in silico methods possibly overestimate the PLD induction effect of cationic amphiphilic compounds compared to the in vitro, with the risk of discarding promising compounds based on incorrect safety liabilities.
KeywordsPhospholipidosis Cationic amphiphilic drugs Toxicophore HepG2 cells Fluorescence assays Organic synthesis
The authors would like to thank Prof. Francesco Galli for generously sharing his laboratory facilities for biological analysis, and Dr. Simon Cross for helpful comments and for English revision. Financial support from the Italian MIUR within the “FIRB-Futuro in Ricerca 2010” Program—Project RBFR10×500 is gratefully acknowledged.
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Conflict of interest
The authors declare that they have no competing interests.
- Chapy H, Goracci L, Vayer P, Parmentier Y, Carrupt PA, Decleves X, Scherrmann JM, Cisternino S, Cruciani G (2015) Pharmacophore-based discovery of inhibitors of a novel drug/proton antiporter in human brain endothelial hCMEC/D3 cell line. Br J Pharmacol 172:4888–4904CrossRefPubMedPubMedCentralGoogle Scholar
- DiMasi JA, Feldman L, Seckler A, Wilson A (2010) Pharmaceutical innovation in the 21st century: new drug approvals in the first decade, 2000–2009. Clin Pharmacol 87:272–277Google Scholar
- Mannhold R, Berellini G, Carosati E, Benedetti P (2006) AOME properties and their prediction major causes for failure in drug development are unsuitable pharmacokinetic properties of drug candidates including absorption, distribution, metabolism, and excretion (ADME), which were traditionally measured at rather late stages of drug development. Nowadays, the testing of ADME properties is done much earlieri. In: Cruciani G (ed) Molecular interaction fields: applications in drug discovery and ADME prediction. Wiley-VCH, New York, 2006, vol. 27 173–196Google Scholar
- Molecular Discovery Ltd. http://www.moldiscovery.com/. Accessed 11 Feb 2016
- Munic V, Banjanac M, Kostrun S, Nujic K, Bosnar M, Marjanovic N, Ralic J, Matijasic M, Hlevnjak M, Erakovic Haber V (2011) Intensity of macrolide anti-inflammatory activity in J774A.1 cells positively correlates with cellular accumulation and phospholipidosis. Pharmacol Res 64:298–307CrossRefPubMedGoogle Scholar
- Ploemen JP, Kelder J, Hafmans T, van de Sandt H, van Burgsteden JA, Salemink PJ, van Esch E (2004) Use of physicochemical calculation of pKa and CLogP to predict phospholipidosis-inducing potential: a case study with structurally related piperazines. Exp Toxicol Pathol 55:347–355PubMedGoogle Scholar
- Sirci F, Goracci L, Rodriguez D, an Muijlwijk-Koezen J, Gutierrez-de-Teran H, Mannhold R (2012) Ligand-, structure- and pharmacophore-based molecular fingerprints: a case study on adenosine A(1), A (2A), A (2B), and A (3) receptor antagonists. J Comput Aided Mol Des 26:1247–1266CrossRefPubMedGoogle Scholar
- SPECS. http://www.specs.net/. Accessed 18 Dec 2015