Design, Synthesis, and Evaluation of Novel ROCK Inhibitors for Glaucoma Treatment: Insights into In Vitro and In Vivo Efficacy and Safety

The inhibition of Rho-associated coiled-coil kinase (ROCK)
has
emerged as a promising strategy for reducing intraocular pressure
(IOP) and treating glaucoma. Here, we report the synthesis and evaluation
of novel ROCK inhibitors, D25 and R3, which
were designed to optimize selectivity, efficacy, and ocular bioavailability. D25 potently inhibited ROCK1/2 with IC50 values
of 47.2 nM and 33.8 nM, respectively, surpassing Netarsudil. Compound R3 had weaker ROCK inhibition but demonstrated favorable lipophilicity
(logP) and good selectivity to ROCKs, which enhances
its potential and safety for ocular delivery. In human trabecular
meshwork (HTM) cells, R3 showed lower cytotoxicity than
Netarsudil and effectively mitigated oxidative damage, enhanced cellular
integrity, and reduced inflammatory cytokine secretion. In rabbit
models, D25 significantly lowered IOP, outperforming
(S)-Netarsudil. R3 exhibited weaker
IOP-lowering efficacy but better selectivity. D25 is
a promising glaucoma treatment candidate, with R3 as
a safer alternative for further optimization.