Neuropathic pain is a prevalent form of chronic pain that is associated with a reduction in glycinergic neurotransmission in the dorsal horn of the spinal cord. Currently available therapeutic treatments for this debilitating condition are either inadequate or elicit severe side effects, such as in the case of opioid analgesics. It has been established that glycinergic neurotransmission can be restored using glycine transporter 2 (GlyT2) inhibitors. ORG25543 is a commercially available and highly potent GlyT2 inhibitor, producing a substantial analgesic response in various rodent models of pain. However, it causes tremors and seizures at higher doses due to irreversible binding. A reversible analogue to ORG25543, termed Compound 1, has proven to be efficacious with a reduced side effect profile, providing proof of concept for reversible GlyT2 inhibitors as a safer analgesic. Another structural analogue, Opiranserin, has inhibitory activity at both GlyT2 and 5HT2A receptors. It is proposed that dual antagonism at GlyT2 and 5HT2A receptors provides effective analgesia through synergistic activity. Opiranserin has progressed through to phase 3 clinical trials, currently being the only GlyT2 inhibitor to reach this juncture. In this study, we compare the in vivo efficacy of these three compounds in a mouse model of neuropathic pain. We also compare potential side effects in a rotarod test of motor function. These results suggest that optimisation of the pharmacokinetics and pharmacodynamics of this class of compounds has the potential to improve their analgesic effects in vivo.