Neuropathic pain affects approximately 10% of the population and is characterised by a lack of glycinergic signalling in the central nervous system which facilitates painful perceptions in abnormal circumstances. Signalling can be normalised by inhibiting glycine transporter-1 (GlyT1) or glycine transporter-2 (GlyT2), to enhance glycinergic signalling and restore normal pain signalling. This study investigates the use of a combined in silico and in vitro approach to generate and develop glycine transport inhibitors to restore the function of glycine neurotransmission as a potential treatment for neuropathic pain. In silico screens using the deep convolutional neural network ‘Atomnet’ generated novel compounds that bind in the lipid allosteric site (LAS) of GlyT2. Phases one and two screens focussed on the head and tail pockets of the LAS respectively. A total of 20 million compounds were screened and the top 80 candidates for each screen were tested for inhibition of GlyT2 expressed in Xenopus laevis oocytes. Two-electrode voltage clamp electrophysiology was used to measure changes in currents to determine the inhibitory action of compounds on GlyT2 and characterise their reversibility and mechanism of inhibition. Two compounds were identified from the phase two screen and in phase 3, a further 94 analogues were tested. Similar studies were used to investigate the activity of lead compounds on closely related GlyT1. 4/94 analogues are moderately potent allosteric GlyT2 inhibitors with promising reversibility properties. Docking simulations confirmed that the compounds are likely to bind in the LAS of GlyT2. The 4 hit compounds also inhibit GlyT1, demonstrating a dual mechanism of action. A combined in silico and in vitro screening approach identified novel compounds that inhibit both GlyT2 and GlyT1. Future in vitro experiments will further explore the molecular basis for drug selectivity and also any inhibitory effects on other closely related transporters. Future in vivo experiments will explore the potential of dual action inhibitors as a treatment for neuropathic pain using animal models.