According to the principle of bioisosterism and the Evans' pharmacophore model of 5-HT3 receptor antagonists, twelve derivatives of tetramethylpyrazine were synthesized. The svructures of the compounds were confirmed by MS, 1H NMR and HRMS. All the target compounds are unreported. The preliminary activity test showed that most of the compounds had fairly potent 5-HT3 receptor anagonistic activities, especially compound 3c. And the conformadonal analysis showed that the active conformation of the compounds was quite fitted to the Evans' pharmacophore model. This indicates that besides the three pharmacophoric elements, the aromatic ring and the substituents attached may affect the bioactivities of the compounds.