Myocardial extractions of pyridaben, a mitochondrial complex I (MC-I) inhibitor, is well correlated with blood flow. Based on the synthesis and characterization of pyridaben analogue 2-tert-butyl-5-[2-(2-[ 18 F]fluroethoxy)ethoxy]benzyloxy]-4-chloro-2H-pyridazin-3-one ([ 18 F]FP2OP), this study assessed its potential to be developed as myocardial perfusion imaging (MPI) agent.Methods: The tosylate labeling precursor 2-(2-(4-(tert-butyl-5-chloro-6-oxo-1,6-dihydro-pyridazin-4-yloxymethyl)benzyloxy)ethoxy)ethyl ester (OTs-P2OP) and the nonradioactive 2-tert-butyl-5-[2-(2-[ 19 F]fluroethoxy)ethoxy]benzyloxy]-4-chloro-2H-pyridazin-3-one ([ 19 F]FP2OP) were synthesized and characterized by IR, 1 H NMR, 13 C NMR and MS analysis. By substituting tosyl of precursor OTs-P2OP with 18 F, the radiolabeled complex [ 18 F]FP2OP was prepared and further evaluated for its in vitro physicochemical properties, in vivo biodistribution, the metabolic stability in mice, ex vivo autoradiography and cardiac PET/CT imaging.Results: Starting with [ 18 F]F − Kryptofix 2.2.2./K 2 CO 3 solution, the total reaction time for [ 18 F]FP2OP was about 100min, with final high-performance liquid chromatography purification included. Typical decay-corrected radiochemical yield stayed at 41±5.3%, the radiochemical purity, 98% or more. Biodistribution in mice showed that the heart uptake of [ 18 F]FP2OP was 41.90±4.52%ID/g at 2min post-injection time, when the ratio of heart/liver, heart/lung and heart/blood reached 6.83, 9.49 and 35.74, respectively. Lipophilic molecule was further produced by metabolized [ 18 F]FP2OP in blood and urine at 30min. Ex vivo autoradiography demonstrates that [ 18 F]FP2OP may have high affinity with MC-I and that can be blocked by [ 19 F]FP2OP or rotenone (a known MC-I inhibitor). Cardiac PET images were obtained in a Chinese mini-swine at 5, 15, 30 and 60min post-injection time with high quality.Conclusion: [ 18 F]FP2OP was synthesized with high radiochemical yield. The promising biological properties of [ 18 F]FP2OP suggest high potential as MPI agent for positron emission tomography in the future.