Justin C Burrell, Tomisin T Latona, Henry Garcia, David R Clizbe, Mykhailo M Tatarchuk, Liwen Zhou, Carlos A Toro, Alexa N Olivarez, Phuong V Nguyen, Cathy Z Yang, George D Bittner, Christopher P Cardozo, D Kacy Cullen
Wallerian degeneration of anucleated axonal segments is driven by SARM1, which depletes axonal NAD+, disrupting energy metabolism and triggering self-destruction. SARM1 inhibition is an emerging therapeutic target for traumatic nerve injuries. Boldine, a natural aporphine alkaloid from Peumus boldus , modulates connexin hemichannels, oxidative stress, and inflammation. Building on our published work showing boldine's neuroprotective effects in nerve injury models, we hypothesized that boldine also inhibits SARM1 directly. A fluorescence polarization assay revealed that boldine inhibits SARM1 NADase activity with an IC50 of approximately 7.5 uM. AI-assisted structural modeling (AlphaFold3-based Boltz-1 with GNINA docking) predicted two boldine binding sites on SARM1: the TIR catalytic site (Kd ∼ 13.5 uM) and the ARM-TIR regulatory interface (Kd ∼ 12 uM). In a sciatic nerve explant model, boldine preserved the integrity of anucleated axonal segments at 3 and 7 days post-transection relative to vehicle controls. These findings suggest boldine may act as a dual-site SARM1 inhibitor and support its development as a neuroprotective therapy after traumatic axonal injury.