Repeated Dosing with NCX1404, a Nitric Oxide-Donating Pregabalin, Re-establishes Normal Nociceptive Responses in Mice with Streptozotocin-Induced Painful Diabetic Neuropathy

Abstract

NCX1404 [(3S)-5-methyl-3-(((1-(4-(nitrooxy)butanoyloxy)ethoxy)carbonylamino) methyl)hexanoic acid] is a novel nitric oxide (NO)-donating pregabalin that is readily absorbed and processed in vivo to pregabalin and NO. We determined the antiallodynic response of NCX1404 after acute or after 7, 14, and 21 days of repeated daily oral dosing in mice with streptozotocin (STZ)-induced painful diabetic neuropathy (PDN). Pregabalin and its combination with the NO donor isosorbide mononitrate (ISMN) were used for comparison. The blood levels of pregabalin and nitrites, used as surrogate marker of NO release, after NCX1404 or pregabalin dosing were monitored in parallel experiments using liquid chromatography with tandem mass spectrometry (LC-MS/MS). NCX1404 and pregabalin resulted in similar pregabalin levels as it was their antiallodynic activity after acute dosing in STZ mice. However, NCX1404 resulted in disease-modifying properties when administered daily for 21 days, as indicated by the time- and dose-dependent reversal of STZ-induced mechanical allodynia (paw withdrawal threshold [PWT]Veh_21d= 1.3 ± 0.15 g for vehicle; PWTNCX1404_21d= 1.4 ± 0.5 g, 2.9 ± 0.2 g* and 4.1 ± 0.2 g*, respectively for 19, 63, and 190μmol/kg, oral gavage [PO] of NCX1404; *P< 0.05 versus vehicle). This effect was not shared by pregabalin at equimolar doses (190μmol/kg, PO, PWTPregab_21d= 1.4 ± 0.1 g*, *P< 0.05 versus equimolar NCX1404). In addition, the NO donor ISMN (52.3μmol/kg, PO) alone or combined with pregabalin (63μmol/kg) was active at 7 days (PWTVeh_7d= 1.7 ± 0.16 g; PWTISMN_7d= 3.9 ± 0.34 g*; PWTPregab_7d= 1.3 ± 0.07 g; PWTISMN+pregab_7d= 3.8 ± 0.29 g*; *P< 0.05) but not at later time points. The long-term effect of NCX1404 was independent of residual drug exposure and lasted for several days after the treatment was stopped. In summary, like pregabalin, NCX1404 is an effective antiallodynic agent. Differently from pregabalin, repeated dosing of NCX1404 re-established normal nociceptive responses in STZ-induced PDN in mice.