To develop a novel and biocompatible copper tetraamine fluoride (CTF) with antibacterial and nondiscolouring properties. This study used copper fluoride and ammonia solution to develop CTF solution. The CTF was characterized by X-ray photoelectron spectroscopy (XPS). Cytotoxicity was evaluated by stem cells from human exfoliated deciduous teeth (SHED) and human gingival fibroblasts (HGF-1). The fluoride concentration was determined using ion-selective electrode. The alkalinity was measured by a pH electrode. The human dentine blocks were treated with CTF and then incubated with Streptococcus mutans (S. mutans) to evaluate the antimicrobial and discolouring effects. The silver diamine fluoride (SDF) was employed as the positive control, and water was the negative control. The colony-forming units (CFUs) and confocal laser scanning microscopy (CLSM) were used to examine the kinetics and viability of the biofilm. The discolouring property on dentine was assessed by spectrophotometry. One-way analysis of variance with the Bonferroni post hoc test was performed to assess and compare the data. XPS confirmed synthesis of CTF solution. The half-maximal inhibitory concentration of CTF on SHED and HGF-1 was 195±16 ppm and 137±11 ppm. The fluoride concentration was 121,000±5,000 ppm. The pH value was 9. Log10 CFU of the CTF, SDF and water group were 5.0±0.2, 4.9±0.1 and 7.4±0.1 (p<0.001, CTF, SDF<Water). CLSM showed that the dead-to-live ratio of the CTF, SDF and water group were 0.8±0.2, 0.8±0.2, and 0.3±0.1 (p<0.001, CTF, SDF>Water). Spectrophotometry showed that the ΔE of the CTF, SDF and water group were 5±2, 6±3 and 45±2 (p<0.001, CTF, Water<SDF). This study developed an alkaline 58% CTF solution, which is biocompatible, antibacterial and non-discolouring. If CTF is successfully translated into clinical care, CTF can be a simple and affordable anti-caries agent for clinicians to prevent dental caries.