NNC 55-0396 [(1S,2S)-2-(2-(N-[(3-Benzimidazol-2-yl)propyl]-N-methylamino)ethyl)-6-fluoro-1, 2,3,4-tetrahydro-1-isopropyl-2-naphtyl cyclopropanecarboxylate dihydrochloride]: A New Selective Inhibitor of T-Type Calcium Channels

Johns Hopkins University, Baltimore, Maryland, United States
Journal of Pharmacology and Experimental Therapeutics (Impact Factor: 3.86). 05/2004; 309(1):193-9. DOI: 10.1124/jpet.103.060814
Source: PubMed

ABSTRACT Mibefradil is a Ca2+ channel antagonist that inhibits both T-type and high-voltage-activated Ca2+ channels. We previously showed that block of high-voltage-activated channels by mibefradil occurs through the production of an active metabolite by intracellular hydrolysis. In the present study, we modified the structure of mibefradil to develop a nonhydrolyzable analog, (1S, 2S)-2-(2-(N-[(3-benzimidazol-2-yl)propyl]-N-methylamino)ethyl)-6-fluoro-1,2,3,4-tetrahydro-1-isopropyl-2-naphtyl cyclopropanecarboxylate dihydrochloride (NNC 55-0396), that exerts a selective inhibitory effect on T-type channels. The acute IC(50) of NNC 55-0396 to block recombinant alpha(1)G T-type channels in human embryonic kidney 293 cells was approximately 7 microM, whereas 100 microM NNC 55-0396 had no detectable effect on high-voltage-activated channels in INS-1 cells. NNC 55-0396 did not affect the voltage-dependent activation of T-type Ca2+ currents but changed the slope of the steady-state inactivation curve. Block of T-type Ca2+ current was partially relieved by membrane hyperpolarization and enhanced at a high-stimulus frequency. Washing NNC 55-0396 out of the recording chamber did not reverse the T-type Ca2+ current activity, suggesting that the compound dissolves in or passes through the plasma membrane to exert its effect; however, intracellular perfusion of the compound did not block T-type Ca2+ currents, arguing against a cytoplasmic route of action. After incubating cells from an insulin-secreting cell line (INS-1) with NNC 55-0396 for 20 min, mass spectrometry did not detect the mibefradil metabolite that causes L-type Ca2+ channel inhibition. We conclude that NNC 55-0396, by virtue of its modified structure, does not produce the metabolite that causes inhibition of L-type Ca2+ channels, thus rendering it more selective to T-type Ca2+ channels.

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    • "To analyze the role played by LT-VACCs in synaptic integration, it is necessary to count on specific blockers devoid of any effect on the high-threshold VACCs that participate in synaptic transmission. The effects of three pharmacological agents known as T-type Ca 2ϩ channel blockers were studied: amiloride (Huguenard and Prince 1992; Tang et al. 1988), Ni 2ϩ (Huguenard and Prince 1992; Lee et al. 1999; Regan 1991), and NNC, a structural analog of mibefradil (Huang et al. 2004; Johnston and Delaney 2010). "
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    ABSTRACT: Low threshold voltage-activated calcium conductances (LT-VACCs) play a substantial role in shaping the electrophysiological attributes of neurites. We have investigated how these conductances affect synaptic integration in a premotor nonspiking (NS) neuron of the leech nervous system. These cells exhibit an extensive neuritic tree, do not fire Na(+)-dependent spikes but express a LT-VACC that was sensitive to 250 µM Ni(2+) and 100 µM NNC 55-0396 (NNC). NS neurons responded to excitation of mechanosensory pressure neurons with depolarizing responses whose amplitude was a linear function of the presynaptic firing frequency. NNC decreased these synaptic responses and abolished the concomitant widespread Ca(2+) signals. Coherent with the interpretation that the LT-VACC amplified signals at the postsynaptic level, this conductance also amplified the responses of NS neurons to direct injection of sinusoidal current. Synaptic amplification thus is achieved via a positive feedback in which depolarizing signals activate a LT-VACC that, in turn, boosts these signals. The wide distribution of LT-VACC could support the active propagation of depolarizing signals, turning the complex NS neuritic tree into a relatively compact electrical compartment. Copyright © 2015, Journal of Neurophysiology.
    Journal of Neurophysiology 05/2015; 114(1):jn.00170.2015. DOI:10.1152/jn.00170.2015 · 3.04 Impact Factor
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    • "However, mibefradil also blocks high-VACCs (Fang and Osterrieder, 1991; Bezprozvanny and Tsien, 1995; Viana et al., 1997). Later on, mibefradil derivative NNC 55-0396 (NNC) emerged as a more selective T-type channel blocker (Huang et al., 2004; Li et al., 2005). Unfortunately, a selective T channel blocker with potential scarce side effects is still an unmet therapeutic goal (Arranz-Tagarro et al., 2014). "
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    ABSTRACT: Adrenal chromaffin cells (CCs) express high-voltage activated calcium channels (high-VACCs) of the L, N and PQ subtypes; additionally, T-type low-VACCs are also expressed during embryo and neonatal life. Effects of the more frequently used T channel blockers NNC 55-0396 (NNC), mibefradil, and Ni2+ on the whole-cell Ba2+ current (IBa), the K+-elicited [Ca2+]c transients and catecholamine secretion, have been studied in adult bovine CCs (BCCs) and rat embryo CCs (RECCs). NNC, mibefradil, and Ni2+ blocked BCC IBa with IC50 of 1.8, 4.9 and 70μM, while IC50 to block IBa in RECCs were 2.1, 4.4 and 41μM. Pronounced blockade of K+-elicited [Ca2+]c transients and secretion was also elicited by the three agents. However, the hypoxia-induced secretion (HIS) of catecholamine in RECCs was blocked substantially (75%) with thresholds concentrations of NCC (IC20 to block IBa); this was not the case for mibefradil and Ni2+ that required higher concentrations to block the HIS response. Thus, out of the three compounds, NNC seemed to be an adequate pharmacological tool to discern the contribution of T channels to the HIS response, without a contamination with high-VACC blockade. Copyright © 2015. Published by Elsevier B.V.
    European Journal of Pharmacology 01/2015; 751. DOI:10.1016/j.ejphar.2015.01.025 · 2.68 Impact Factor
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    • "rt and others J Physiol 592 . 1 of mibefradil ( stratum lucidum stimulation : 1 . 1 ± 4 . 9% IPSC amplitude reduction , n = 7 ; stratum granulosum stimulation : 4 . 7 ± 2 . 9% increase , n = 9 ; P > 0 . 4 ) ( Fig . 2B ) . Finally , we applied the potent inhibitor of T - type Ca 2+ channels containing Ca v 3 . 1 subunits ( NNC 55 - 0396 , 10 μM ) ( Huang et al . 2004 ) and found similar results ( stratum lucidum stimulation : 2 . 7 ± 7 . 7% IPSC amplitude increase ; stratum granulosum stimulation : 8 . 3 ± 3 . 0% decrease , n = 7 ; P > 0 . 1 ) ( Fig . 2C ) . Inter - estingly , T - type Ca 2+ channel blockers reduced the amplitude of evoked IPSCs to approximately similar levels ( NiCl 2 , stratum luc"
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    ABSTRACT: Zinc actions on synaptic transmission span the modulation of neurotransmitter receptors, transporters, activation of intracellular cascades or alterations in gene expression. Whether and how zinc affects inhibitory synaptic signalling in the dentate gyrus remains largely unexplored. We found that mono- and di-synaptic GABAergic inputs onto dentate granule cells were reversibly depressed by exogenous zinc application and enhanced by zinc chelation. Blocking T-type Ca2+ channels prevented the effect of zinc chelation. When recording from dentate fast-spiking interneurones, zinc chelation facilitated T-type Ca2+ currents, increased action potential half-width and decreased spike threshold. It also increased the offset of the input-output relation consistent with enhanced excitability. In granule cells, chelation of zinc reduced the time window for integration of glutamatergic inputs originating from perforant-path synapses resulting in reduced spike transfer. Thus, zinc-mediated modulation of dentate interneurone excitability and GABA release regulates information flow to local targets and hippocampal networks.
    The Journal of Physiology 09/2013; 592(1). DOI:10.1113/jphysiol.2013.261420 · 4.54 Impact Factor
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