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.
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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

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Figure 12

C2230 does not affect sensitivity to mechanical, cold, or nociceptive heat stimulation, nor motor function.

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C2230 does not affect sensitivity to mechanical, cold, or nociceptive he...
(A) Left: Schematic representation and timeline of the experimental approach used to assess mechanical threshold and aversion time in naive mice following C2230 administration. Right: Schematic representation and timeline of the experimental approach used to evaluate thermal stimulation responses and motor function in naive mice 2 hours after C2230 administration. (B and F) Time course of von Frey mechanical thresholds after i.p. administration of vehicle or C2230 in naive male and female mice. (C and G) Time course of aversion time responses to an acetone drop following i.p. administration of vehicle or C2230 in naive male and female mice. P values as indicated; 2-way ANOVA followed by Bonferroni’s multiple comparison test; n = 8 mice per group. (D and H) Bar graphs showing withdrawal latency to a 52°C nociceptive stimulus in male and female naive mice 2 hours after C2230 injection. (E and I) Bar graphs representing latency to fall in the rotarod test for male and female naive mice 2 hours after C2230 injection. P values as indicated, Mann Whitney test; n = 8 mice per group. Values are expressed as mean ± SEM. See Supplemental Table 3 for full statistical analysis.