Fr173657

The peptides bradykinin [BK; H-Arg¹-Pro²-Pro³-Gly⁴-Phe⁵-Ser⁶-Pro⁷-Phe⁸-Arg⁹-OH)], NG291 ([Hyp³,Thi⁵,^NChg⁷,Thi⁸]-BK; B2R agonist), HOE140 (dArg[Hyp³,Thi⁵,dtic⁷,Oic⁸]-BK; B2R antagonist), and NG29 (SarLys[dphe⁸]desArg⁹-BK; B1R agonist) were assembled on a solid support by an automated Pioneer peptide synthesizer using 9-fluorenylmethyoxy-carbonyl chemistry (54 (link), 55 (link)). Peptides were purified by analytical reversed-phase high-performance liquid chromatography (RP-HPLC) and were >95% pure, with expected mass spectra. The peptide B2R agonist RMP-7 ([Hyp³,Thi⁵,(4-Me)Tyr⁸(ΨCH₂NH)Arg⁹]-BK) was supplied by Bachem Bioscience. Peptides were stored in the powder form at −20 °C. Stock solutions (10 mM) of peptides were also prepared in Nanopure water. The nonpeptide B2R antagonist FR173657 was kindly provided by Astellas Pharma Inc (formerly Fujisawa Pharmaceutical Co) and was prepared in 50% dimethyl sulfoxide (DMSO)/50% Nanopure water.

SSR240612 (Sanofi Aventis) was used at 10 μM to block B1R and FR173657 (Astellas Pharma) at 10 μM to block B2R. The agonist desArg9-Bradykinin (DABK) (Sigma-Aldrich) at 0.1 μM was used to activate B1R. Bradykinin fragment 1–8 (BK) (Sigma-Aldrich) at 0.1 μM was used to activate B2R.