Olfactory receptor164 (Olfr164) belongs to the G-protein coupled receptor (GPCR) family. It is one of the ORs expressed on neutrophils in mice and a potential therapeutic target for sepsis caused by Pseudomonas aeruginosa infection. However, the development of selective GPCR-targeting drugs is still challenging due to the complexity of GPCR dynamic features, the structural similarity between members of the GPCR family, and highly conserved residues in the orthosteric sites of GPCR. We identified that chromone-2-carboxamide derivatives could act as ligands for the Olfr164 receptor by GPCR library screening. Herein, we have rationally designed and synthesized a novel series of amide derivatives for the Olfr164 receptor. The chromone analogues were synthesized by acylation of 2-hydroxyacetophenones with diethyl oxalate, acid-catalyzed cyclization, and ester hydrolysis in 21-84% overall yield two steps. The synthesis of amine derivatives was achieved in 18-44% yield via two steps: Strecker reaction of the aldehyde, and reduction of the cyanide with LAH. Among the synthesized compounds, five compounds (KB-4814, 4824, 4829, 4839, and 4844) were identified as Olfr164 ligands at in vitro cAMP assays. Overall, the five compounds could be used as a potential hit compound for the further development of novel Olfr164 ligands.