Probenecid-D14

Probenecid-D14 (4[(Dipropylamino-d14)sulfonyl]benzoic Acid) is a deuterium-labeled isotope of Probenecid (T0457) and can be used for isotope tracing. Probenecid, a benzoic acid derivative, exhibits antihyperuricemic properties and selectively activates the transient receptor potential vanilloid receptor 2 (TRPV2) while also inhibiting the pannexin 1 channel.

Method: In Sf9 insect cell membrane vesicles expressing MRP1 or MRP2, 4 μM N-ethylmaleimide glutathione (NEM-GS) was added as a substrate, along with varying concentrations of probenecid. ATP-dependent NEM-GS uptake was measured using a rapid filtration method. In the same membrane vesicles, the effect of probenecid on MRP1 and MRP2 ATPase activities was assessed by measuring vanadate-sensitive ATPase activity.
Result: Probenecid inhibited NEM-GS uptake in both MRP1 and MRP2. Regarding ATPase activity, probenecid primarily inhibited MRP1 but significantly stimulated MRP2 ATPase activity, with maximal stimulation observed at approximately 2 mM, exhibiting a bell-shaped curve. In the presence of GSH, probenecid further increased MRP2 ATPase activity [1].
Method: A GPCR calcium mobilization assay was used to evaluate the inhibitory effects of probenecid on the bitter taste receptor subtypes hTAS2R16, hTAS2R38, and hTAS2R43. Cells were pre-incubated with probenecid prior to receptor activation to determine whether probenecid exerts its inhibitory effects via an allosteric mechanism.
Result: Probenecid specifically inhibited hTAS2R16-mediated cellular responses, but had no significant effect on hTAS2R38 or hTAS2R43. Its mechanism of action was determined to be non-competitive allosteric inhibition, providing evidence for a direct interaction between probenecid and GPCRs. Furthermore, probenecid can be used to enhance cellular signaling in GPCR calcium mobilization assays [2].

Method: Wild-type mice were injected with various doses of Probenecid (2–200 mg/kg) via the tail vein. Cardiac function was assessed using echocardiography (M-mode and B-mode). Ejection fraction (EF) and fractional shortening (FS) were measured, and a dose-response curve was generated.
Result: Probenecid dose-dependently increased the ejection fraction in wild-type mice, with an EC50 of approximately 49.33 mg/kg. The effect occurred within 5 minutes after administration and lasted for at least 1 hour [1].
Method: Wild-type mice were intravenously injected with Probenecid (30 mg/kg and 100 mg/kg). A pressure catheter was inserted into the right carotid artery to monitor changes in left ventricular +dP/dt.
Result: Probenecid rapidly increased +dP/dt at both 30 mg/kg and 100 mg/kg, indicating an enhancement of myocardial contractility [1].