title: In vitro inflammatory/anti-inflammatory effects of nitrate esters of purines.
abstract: Six purine analogues bearing a nitrate ester group (potential NO donor) were tested on human THP-1 macrophages to investigate their effects on the inflammatory response. Only three analogues increased the basal level of IL-1β. Two analogues exacerbated the inflammatory response induced by ATP but not that induced by H2O2. Only 6-[4-(6-nitroxyacetyl)piperazin-1-yl]-9H-purine (compound MK128) abolished ATP or H2O2-induced IL-1β production in the culture medium. Similar results were reproduced on macrophages differentiated from buffy coats and stimulated with LPS. MK128 was the only analogue to release NO and leading to nitrite formation in the culture medium. The EC50 for inhibition of induced IL-1β production by the cells was estimated to be 10-12µg/ml (about 36µM) and corresponded to the production of around 30µM nitrites in the culture medium. This anti-inflammatory effect of MK128 was mimicked by trinitrin used in 10 fold higher concentrations. Preincubation of cells with NO trapper cPTIO partially abolished the beneficial effect of MK128 while MK137, a ONO2 deprived analogue of MK128, was not able to inhibit induced IL-1β production and proved to be inflammatory. Moreover, purinergic channel inhibitors (oATP and U73122) inhibited the MK137 inflammatory effect. Finally, MK128 reduced the quantity of p20 caspase-1 produced in the culture medium. We suggest that MK128 inhibits IL-1β production via NO production and subsequent inflammasome component nitrosylation. On the opposite MK137, deprived from ONO2 group, could act as agonist of purinergic receptors and could thus activate inflammasome.
authors.affiliation: INSERM U866, LPPCM, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France; Cohiro Biotechnology, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France.
authors.forename: Loranne
authors.initials: L
authors.lastname: Maugé
authors.affiliation: Institute of Biology, Medicinal Chemistry and Biotechnology, National Hellenic Research Foundation, 48 Vas. Constantinou Ave., 11635 Athens, Greece.
authors.forename: Theano
authors.initials: T
authors.lastname: Fotopoulou
authors.affiliation: Cohiro Biotechnology, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Delemasure
authors.affiliation: INSERM U866, LPPCM, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France; Cohiro Biotechnology, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Dutartre
authors.affiliation: Institute of Biology, Medicinal Chemistry and Biotechnology, National Hellenic Research Foundation, 48 Vas. Constantinou Ave., 11635 Athens, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Koufaki
authors.affiliation: INSERM U866, LPPCM, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France; Cohiro Biotechnology, Faculté de Médecine, 7 bvd Jeanne d'Arc, 21000 Dijon, France. Electronic address: connatjl@u-bourgogne.fr.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Connat
mesh_terms: D000893:Anti-Inflammatory Agents / Q000009:adverse effects / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D020170:Caspase 1 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004952:Esters; D006801:Humans; D007249:Inflammation / Q000188:drug therapy; D053583:Interleukin-1beta / Q000096:biosynthesis; D009566:Nitrates / Q000737:chemistry*; D009569:Nitric Oxide / Q000096:biosynthesis; D011687:Purines / Q000009:adverse effects / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D018048:Receptors, Purinergic P2 / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D004952:Esters; D053583:Interleukin-1beta; D009566:Nitrates; D011687:Purines; D018048:Receptors, Purinergic P2; D009569:Nitric Oxide; D020170:Caspase 1; C030985:purine
keywords: ATP; Caspase-1; H(2)O(2); Interleukin-1β; NO donor; THP-1