Background and Purpose: Adenosine is a local mediator that regulates physiological and pathological processes via activation of four GPCRs (A1, A2A, A2B, and A3). We have investigated the effect of two A1-receptor-selective agonists and the novel A1-receptor bitopic ligand VCP746 on the rat cardiovascular system. Experimental Approach: The regional haemodynamic responses of these agonist was investigated in conscious rats. Male Sprague–Dawley rats (350–450 g) were chronically implanted with pulsed Doppler flow probes on the renal, mesenteric arteries and the descending abdominal aorta and the jugular vein and caudal artery catheterized. Cardiovascular responses were measured following intravenous infusion (3 min each dose) of CCPA (120, 400, and 1,200 ng·kg−1·min−1), capadenoson or adenosine (30, 100, and 300 μg·kg−1·min−1), or VCP746 (6, 20, and 60 μg·kg−1·min−1) following pre-dosing with DPCPX (0.1 mg·kg−1, i.v.) or vehicle. Key Results: CCPA produced a significant A1-receptor-mediated decrease in heart rate that was accompanied by vasoconstrictions in the renal and mesenteric vascular beds but an increase in hindquarters vascular conductance. The partial agonist capadenoson also produced an A1-receptor-mediated bradycardia. In contrast, VCP746 produced increases in heart rate and renal and mesenteric vascular conductance that were not mediated by A1-receptors. In vitro studies confirmed that VCP746 had potent agonist activity at both A2A- and A2B-receptors. Conclusions and Implications: These results suggest VCP746 mediates its cardiovascular effects via activation of A2 rather than A1 adenosine receptors. This has implications for the design of future bitopic ligands that incorporate A1 allosteric ligand moieties.