C2230, a preferential use- and state-dependent CaV2.2 channel blocker, mitigates pain behaviors across multiple pain models
Cheng Tang, … , Olga A. Korczeniewska, Rajesh Khanna
Cheng Tang, … , Olga A. Korczeniewska, Rajesh Khanna
Published December 10, 2024
Citation Information: J Clin Invest. 2025;135(4):e177429. https://doi.org/10.1172/JCI177429.
View: Text | PDF
Research Article Neuroscience

C2230, a preferential use- and state-dependent CaV2.2 channel blocker, mitigates pain behaviors across multiple pain models

Abstract

Antagonists — such as Ziconotide and Gabapentin — of the CaV2.2 (N-type) calcium channels are used clinically as analgesics for chronic pain. However, their use is limited by narrow therapeutic windows, difficult dosing routes (Ziconotide), misuse, and overdoses (Gabapentin), as well as a litany of adverse effects. Expansion of novel pain therapeutics may emerge from mechanism-based interrogation of CaV2.2. Here, we report the identification of C2230, an aryloxy-hydroxypropylamine, as a CaV2.2 blocker. C2230 trapped and stabilized inactivated CaV2.2 in a slow-recovering state and accelerated the open-state inactivation of the channel, conferring an advantageous use-dependent inhibition profile. C2230 inhibited CaV2.2 during high-frequency stimulation, while sparing other voltage-gated ion channels. C2230 inhibited CaV2.2 in dorsal root and trigeminal ganglia neurons from rats, marmosets, and humans in a G-protein-coupled-receptor–independent manner. Further, C2230 reduced evoked excitatory postsynaptic currents and excitatory neurotransmitter release in the spinal cord, leading to relief of neuropathic, orofacial, and osteoarthritic pain-like behaviors via 3 different routes of administration. C2230 also decreased fiber photometry-based calcium responses in the parabrachial nucleus, mitigated aversive behavioral responses to mechanical stimuli after neuropathic injury, and preserved protective pain responses, all without affecting motor or cardiovascular function. Finally, site-directed mutation analysis demonstrated that C2230 binds differently than other known CaV2.2 blockers, making it a promising lead compound for analgesic development.

Authors

Cheng Tang, Kimberly Gomez, Yan Chen, Heather N. Allen, Sara Hestehave, Erick J. Rodríguez-Palma, Santiago Loya-Lopez, Aida Calderon-Rivera, Paz Duran, Tyler S. Nelson, Siva Rama Raju Kanumuri, Bijal Shah, Nihar R. Panigrahi, Samantha Perez-Miller, Morgan K. Schackmuth, Shivani Ruparel, Amol Patwardhan, Theodore J. Price, Paramjit S. Arora, Ravindra K. Sharma, Abhisheak Sharma, Jie Yu, Olga A. Korczeniewska, Rajesh Khanna

×

Figure 1

Identification of the aryloxy-hydroxypropylamine compound C2230 as a preferential CaV2.2 channel antagonist.

Options: View larger image (or click on image) Download as PowerPoint
Identification of the aryloxy-hydroxypropylamine compound C2230 as a pre...
(A) Chemical structure of a racemic mixture of C2230. (B) Typical current traces from CaV2.2-expressing (Rattus norvegicus) HEK293 cells in the presence and absence of 5 μM C2230 at the holding potentials (Vh) of –50 mV and –80 mV (n = 10–16 cells). (C) Dose-response relationships of C2230 inhibiting CaV2.2 currents at the 2 Vh (n = 12–14 cells). (D and E) Time-course of C2230 inhibiting the CaV2.2 currents and subsequent recovery upon compound washing off (E), the typical traces in D represent the currents at time points of 1, 2, and 3, as indicated in E. Perfusion of 0.1% DMSO served as the negative control (n = 7–8 cells). (F and G) Dose-response relationships of C2230 inhibiting the heterologously expressed KV2.1, NaV1.5, CaV1.2, CaV3.1, CaV3.2, and CaV3.3 channels, with the IC50s being determined as 28.0 ± 5.4 μM and 65.8 ± 12.2 μM for KV2.1, 8.7 ± 1.0 μM and 18.1 ± 3.5 μM for NaV1.5, 22.7 ± 6.3 μM and 26.9 ± 6.0 μM for CaV1.2, 9.2 ± 1.6 μM and 7.6 ± 0.9 μM for CaV3.1, 9.9 ± 1.7 μM and 8.3 ± 2.5 μM for CaV3.2, and 13.5 ± 2.2 μM and 10.5 ± 1.3 μM for CaV3.3, at the Vh of –80 mV (F) and –50 mV (G), respectively (n = 5–9 cells). The CaV2.2 curves were included for comparison. All data are from at least 3 independent experiments.