Establishment Of HEK293 Cell Lines Stably Expressing NPS Receptor And The Screening Of High Efficient Agonists

Neuropeptide S(NPS) receptor display widespread localization in the central nervous system, and the NPS/NPSR system, potently modulates wakefulness, anxiety, arousal, locomotion, food intake, memory, and drug addiction. NPSR activation will increases in intracellular calcium levels. Structure-activity studies demonstrated that:(1) The NPS(1-10) fragment is the shortest to remain the function of activate NPSR, and the C-terminal 11-20 sequence is necessary for its in vivo biological effects; (2) The N-terminal Phe2-Arg3-Asn4 sequence of NPS is very important for NPSR recognition, the sequence Gly5-Val6-Gly7 is seems to be important to maintain bioactive conformation of peptide. The best analog produced peptides with less than 2-fold increased potency in these structure-activity studies on NPS. In our experiments, we established HEK293 cells which could stably express NPSR and 50 NPS analogues were synthesized to found better NPSR agonists. Replacement of Ser1with isobutyric acid displayed a potency value increased significantly that of NPS. The best compound of our studies, analog 48, was 7.6-fold higher pontecy than the natural peptide. Finally, we demonstrated that analog 48, like NPS, at dose able to increase locomotor activity and evoked anxiolytic-like effects in mice.