Additional file 2 of Efficacy and safety of GV1001 in patients with moderate-to-severe Alzheimer’s disease already receiving donepezil: a phase 2 randomized, double-blind, placebo-controlled, multicenter clinical trial

Additional file 2: Table S1. Inclusion and exclusion criteria of this clinical trial. Table S2. Participant number of each group. Table S3. Observed mean total score of each outcome. Table S4. Summary of CIBIS and CIBIC-plus distributions in full analysis set. Table S5. Least mean difference from placebo in full analysis set. Table S6. Least mean difference from placebo in per-protocol population. Table S7. Overall summary of treatment-emergent adverse events: safety set population. Table S8. Most frequent treatment-emergent adverse events that occurred in > 2 patient overall: safety set population. Figure S1. GV1001 (1 mg/kg) or an equivalent volume of 0.9% saline was subcutaneously injected into old 3xTg-AD mice (B6:129-Psen1tm1Mpm Tg[APPSwe, tauP301L]1Lfa/Mmjax) from the age of 21 months until the mice were deemed ready for sacrifice according to the CCAC guidelines on selecting an appropriate endpoint in experiments using animals for research, teaching, and testing. The injections were administered three times a week until the endpoint. The neurobehavioral functions were evaluated every 3 days until the endpoint using the Y-maze test and passive avoidance task. A. In the Y-maze test, which was used to measure the willingness of rodents to explore new environments and therefore quantify the cognitive deficits,1,2 compared with 0.9% saline, 1 mg/kg GV1001 significantly improved the percentage of spontaneous alternations among the Y-maze arms. B and C. In the passive avoidance tasks, which is a fear-aggravated test used to evaluate learning and memory, the mice received an electric shock when they entered the dark compartment3,4; therefore, the latency to enter the dark compartment and number of errors reflected the learning and memory of the mice. The latency to enter the dark compartment and number of errors were significantly improved following treatment with 1 mg/kg GV1001 (B and C, respectively). D. To confirm that GV1001 can enter the brain, 1 mg/kg of GV1001 conjugated with ferrocenecarboxylic acid, which we used in our previous study,5 was subcutaneously injected into 3xTg-AD (12-month-old) mice. It was detected as dark signals in the brain using 3 T magnetic resonance imaging (white arrows) and Prussian blue staining (black boxes).