title: In vitro and in silico analysis of the vascular effects of asymmetrical N,N-bis(alkanol)amine aryl esters, novel multidrug resistance-reverting agents.
abstract: Asymmetrical N,N-bis(alkanol)amine aryl esters (FRA77, GDE6, and GDE19) are potent multidrug resistance (MDR) reversers. Their structures loosely remind that of the Ca(2+) antagonist verapamil. Therefore, the aim of this study was to investigate their vascular activity in vitro. Their effects on the mechanical activity of fresh and cultured rat aorta rings on Cav1.2 channel current (I Ca1.2) of A7r5 cells and their cytotoxicity on A7r5 and EA.hy926 cells were analyzed. Docking at the rat α1C subunit of the Cav1.2 channel was simulated in silico. Compounds tested were cytotoxic at concentrations >1 μM (FRA77, GDE6, GDE19) and >10 μM (verapamil) in EA.hy926 cells, or >10 μM (FRA77, GDE6, GDE19) and at 100 μM (verapamil) in A7r5 cells. In fresh rings, the three compounds partly antagonized phenylephrine and 60 mM K(+) (K60)-induced contraction at concentrations ≥1 and ≥3 μM, respectively. On the contrary, verapamil fully relaxed rings pre-contracted with both agents. In cultured rings, 10 μM GDE6, GDE19, FRA77, and verapamil significantly reduced the contractile response to both phenylephrine and K60. Similarly to verapamil, the three compounds docked at the α1C subunit, interacting with the same amino acids residues. FRA77, GDE6, and GDE19 inhibited I Ca1.2 with IC50 values 1 order of magnitude higher than that of verapamil. FRA77-, GDE6-, and GDE19-induced vascular effects occurred at concentrations that are at least 1 order of magnitude higher than those effectively reverting MDR. Though an unambiguous divergence between MDR reverting and vascular activity is of overwhelming importance, these findings consistently contribute to the design and synthesis of novel and potent chemosensitizers.
authors.affiliation: Dipartimento di Scienze della Vita, Università degli Studi di Siena, Via A. Moro 2, 53100, Siena, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Fusi
authors.affiliation: Dipartimento di Scienze della Vita, Università degli Studi di Siena, Via A. Moro 2, 53100, Siena, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Durante
authors.affiliation: Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, via A. Moro 2, 53100, Siena, Italy.
authors.forename: O
authors.initials: O
authors.lastname: Spiga
authors.affiliation: Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, via A. Moro 2, 53100, Siena, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Trezza
authors.affiliation: Dipartimento di Scienze della Vita, Università degli Studi di Siena, Via A. Moro 2, 53100, Siena, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Frosini
authors.affiliation: Dipartimento NEUROFARBA-Sezione di Farmaceutica e Nutraceutica, Università di Firenze, via Ugo Schiff 6, 50019, Sesto Fiorentino, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Floriddia
authors.affiliation: Dipartimento NEUROFARBA-Sezione di Farmaceutica e Nutraceutica, Università di Firenze, via Ugo Schiff 6, 50019, Sesto Fiorentino, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Teodori
authors.affiliation: Dipartimento NEUROFARBA-Sezione di Farmaceutica e Nutraceutica, Università di Firenze, via Ugo Schiff 6, 50019, Sesto Fiorentino, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Dei
authors.affiliation: Dipartimento di Scienze della Vita, Università degli Studi di Siena, Via A. Moro 2, 53100, Siena, Italy. simona.saponara@unisi.it.
authors.forename: S
authors.initials: S
authors.lastname: Saponara
mesh_terms: D000588:Amines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D002121:Calcium Channel Blockers / Q000378:metabolism / Q000494:pharmacology*; D020746:Calcium Channels, L-Type / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D018432:Drug Resistance, Multiple / Q000187:drug effects*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D004952:Esters / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008297:Male; D008564:Membrane Potentials; D062105:Molecular Docking Simulation; D015394:Molecular Structure; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D013329:Structure-Activity Relationship; D046509:Tissue Culture Techniques; D014664:Vasodilation / Q000187:drug effects*; D014665:Vasodilator Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D014700:Verapamil / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000588:Amines; C504728:Cacna1c protein, rat; D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D004952:Esters; D014665:Vasodilator Agents; D014700:Verapamil
keywords: A7r5 cells; Cav1.2 channel current; Docking; EA.hy926 cells; MDR reverter; Rat aorta rings