EED-IN-5 is an orally active, EZH2-selective trisubstituted pyridine EED-H3K27me3 inhibitor and immunomodulator with anti-inflammatory properties. It targets EED with an IC50 of 28.21 nM. In mouse models, oral administration of EED-IN-5 leads to preferential and sustained accumulation in lymph nodes. This compound reduces H3K27me3 levels in dendritic cells and inhibits their migration, consequently decreasing the infiltration of specific dendritic cells, macrophages, and T cells into the spinal cord and brain. EED-IN-5 shows hERG inhibition activity, tests negative in Mini-Ames assays, and does not exhibit significant toxicity with long-term high-dose administration. EED-IN-5 is applicable for multiple sclerosis research.