Proc. West. Pharmacol. Soc.

50: 00-00 (2007)

Vasoactive Properties of Antidepressant N-Alkyl Derivatives

Renata Snchez-Ortiz1, Vctor Prez-lvarez1 and Rosa A. Bobadilla Lugo2*
2

Seccin Externa de Farmacologa, Centro de Investigacin y de Estudios Avanzados del IPN Av. IPN 2508, Zacatenco, Mxico; Departamento de Fisiologa y Farmacologa, Escuela Superior de Medicina, Instituto Politcnico Nacional, Plan de San Luis y Daz Mirn, Casco de Santo Toms, Mxico *E-mail: rabobadilla@terra.com.mx

ABSTRACT Hypotension is a principal side effect of antidepressant therapy. In addition to serotonin and noradrenalin reuptake inhibition, some antidepressants have shown ion channel interactions which are thought to be related to the vascular effects of these agents. Methylation of the pharmacophore has shown to change the pharmacological properties of a variety of compounds. The purpose of this work was to evaluate whether methylation of the amino group of imipramine (TCAs) and fluoxetine (SSRI) could change their vasodilator properties. N-methyl imipramine (NMI), N-methyl fluoxetine and (NMF) NN dimethyl fluoxetine (NNDF) were synthesized and compared with desipramine (DES), imipramine (IMI) and fluoxetine (F) in their ability to relax rat aortic rings pre-contracted with 80mM KCl using an isolated bath preparation. Drugs were evaluated in thoracic aorta rings with and without endothelium. All compounds displayed a vasorelaxant effect. Endothelium-denuded aortic rings showed an increased relaxant response for IMI and derivatives compared with endothelium-intact vessels, while no endothelium-dependent effect was observed with F and its methyl derivatives. Maximal relaxant potency was displayed by dimethylated derivatives (IMI and NMF), while NMI in the TCA series and NNDF in the SSRI series (both with 3 methyl groups), had the least potency to relax either preparation. Endothelium plays an important role inhibiting the vasodilatation induced by IMI and its derivatives. Vascular relaxation is increased in the compounds tested with 2 methyl groups in their structure, while the presence of 3 methyl groups (positive charge) importantly reduced the relaxant potency. INTRODUCTION In addition to their principal antidepressant effects attributed to their ability to inhibit presynaptic monoamine recapture, tryciclic antidepressants and 5HT selective fluoxetine recapture inhibitors are associated with adverse autonomic responses related to their autonomic properties. Cardiovascular effects include sinus tachycardia, and variable prolongation of cardiac conduction times with potential for arrhythmias, particularly with overdose. In the absence or cardiac disease, the main vascular

problem associated with imipramine like agents is postural hypotension, which can precipitate falls and injuries [1]. Vasodilation has been related to anti-1 adrenergic actions, but other mechanisms, including transmembrane ion flux, could also be involved [2]. In addition, fluoxetine has been reported to antagonize nicotinic acetylcholine and 5-HT receptors and to block Na+ and voltage-dependent K+ channels in neuronal tissue [3]. Wermuth in 1996 [5] found that the replacement of a hydrogen atom by a substituent (alkyl, halogen, hydroxyl, nitro, cyano, alkoxy, amino, carboxylate, etc.) in an active molecule, can significantly modify the potency, duration of action, and perhaps even the nature of the pharmacologic effect. The purpose of this work was to evaluate whether methylation of the amino group of imipramine (TCA) and fluoxetine (SSRI) could change their vasodilator properties in rings from rat thoracic aorta.
METHODS N-metyl imipramine (NMI), N-methyl fluoxetine and (NMF) N-N dimethyl fluoxetine (NNDF) were methylated by the method of Garca-Ferreiro et al., 2004 [4]. Arterial segments were taken from young male Wistar rats (200-250 g). Animals were anesthetized with ether and thoracic aorta segments were dissected free of connective tissue, placed in Petri dishes containing Krebs buffer and were cut in rings of 4-5 mm; tissues were mounted in organ bath containing 10 ml de Krebs solution of the following mM composition: NaCl, 118; KCl, 5.0; MgCl2.6H2O, 1.2; CaCl2, 3.0; NaHCO3, 25; glucose, 11.1; K2HPO4, 0.9; EDTA, 0.03.The temperature was maintained at 37 C and the preparation were aerated with 95% de O2 y 5% de CO2. Experimental groups were divided in thoracic aorta rings with and without endothelium. To study relaxation, arterial rings were contracted with KCl (80 mM). After a stable contraction was obtained, relaxant effects of to N-metil imipramine (NMI), Nmethyl fluoxetine and (NMF) N-N dimethyl fluoxetine (NNDF) were compared with the elicited by desipramine (DES), -8 -4 imipramine (IMI) and fluoxetine (F) (10 M 10 M), in a cumulative manner.

RESULTS Effects of TCAs on pre-contracted aortic rings: The TCAs DES, IMI and their derivative NMI induced a dose-dependant relaxation of KCl pre-contracted vascular rings compared with vehicle (DMSO) in both experimental groups (Fig. 1A and B for rat aortic rings with and without endothelium, respectively). In both

preparations, NMI elicited less vasodilation and potency in comparison with DES and IMI. The relaxant responses were markedly increased in the aortic rings without endothelium with a similar pattern than the endothelium-denuded rings with the IMI derivatives.
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DISCUSSION Structure-activity relationship studies employing substituent modifications represent common practice in medicinal chemistry. As many as half of existing drugs on the market contain easily substituted aromatic rings. In this work we methylated imipramine, fluoxetine and N-methyl fluoxetine and obtained a series of mono-, di- and tri-methylated derivatives: desipramine (DES), fluoxetine (F), imipramine (IMI), N-methyl fluoxetine (NMF), N-metyl imipramine (NMI), and N-N dimethyl fluoxetine (NNDF) and compared their ability to relax rat aortic rings pre-contracted with KCl in an isolated tissue bath preparation. We observed relaxant potency of 2 > 1 > 3 with regard to the number of methyl substituents. In this work, substitution with 2 methyl groups seems to represent the ideal biological result that is independent of a steric effect. On the other hand, the reduced effects of tri-methylated derivatives may be attributable to their positive charge, inhibiting drugreceptor or drug-channel interactions. Similar relaxant potency was observed for both groups of compounds, suggesting they share a similar vasorelaxant mechanism. Nevertheless, tricyclic but not F derivatives, showed increased relaxant effects on the endothelium denuded thoracic aorta, suggesting that tricyclic derivatives evoke an additional mechanism in the endothelium that inhibits its effects in vascular smooth muscle. Then, tri-methylated IMI or F derivatives would potentially produce less hypotension than the parent compounds. The hypotensive effects of IMI derivatives are partially blunted by the vascular endothelium. Nevertheless contribution of the other pharmacokinetic and pharmacodynamic factors should also be taken into account.
ACKNOWLEDGMENTS Supported by CONACYT 176691 grant number.

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Figure 1A. Vasodilation of IMI and derivatives in rat aortic rings with endothelium. DES ( , n=7); IMI ( , n=7); NMI ( , n=7); DMSO ( , n=8). B. Vasodilation of IMI and derivatives in rat aortic rings without endothelium. DES ( , n=8); IMI ( , n=8); NMI ( , n=8); DMSO ( , n=5). Values are mean SEM, *p<0.050.

Effects of SSRI and derivatives on pre-contracted artery rings: For the series of SSRI, F and their derivatives, NMF and NNDF, they too induced a dose-dependant relaxation of pre-contracted vascular rings in both experimental groups (Fig. 2A and B; rat aortic rings with and without endothelium, respectively). In both preparations, NNDF elicited less vasodilation and potency compared to F and NMF, in the same manner as NMI. In this series of compounds, there are no differences in the maximum effect of dilation elicited in presence or absence of endothelium.
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Figure 2A. Vasodilation of F and derivatives in rat aortic rings with endothelium. F ( , n=8); NMF ( , n=8); NNDF ( , n=8); DMSO ( , n=7). B. Vasodilation of F and derivatives in rat aortic rings without endothelium. F ( , n=12); NMF ( , n=6); NNDF ( , n=11); DMSO ( , n=10). Values are mean SEM, *p<0.050.

REFERENCES 1. Baldessarini, R.J. (2005) Ch. 17. Brunton, Lazo, and Parker, Ed. Goodman & Gilmans The Pharmacological Basis of the Therapeutics, 11 Ed., McGraw-Hill, NY. 2. Becker, B., Morel, N., et al. (2004) Biochem Pharmacol .68, 833-842. 3. Ungvari, Z., Pacher, P., et al. (1999) Stroke. 30,19491954. 4. Garca-Ferreiro, R.E., Kerzchensteiner, D., et al. (2004) J. Gen. Physiol. 124, 301-317. 5. Wermuth CG. (2001) Specific Substituent effects. The Practice of Medicinal Chemistry. 4 Ed., Academic Press, San Diego.