Reparixin

Binding assays

Isolated PMNs (107×mL) were resuspended in RPMI 1640 and incubated at 37℃ for 15 min in the presence of repertaxin (1 mM) or vehicle. After incubation cells were resuspended (2×107/mL) in binding medium (RPMI 1640 containing 10 mg/ml BSA, 20 mM HEPES, and 0.02% NaN3) in the presence of repertaxin or vehicle. Aliquots of 0.2 nM of [125I]CXCL8 and serial dilutions of unlabeled CXCL8 were added to 106 cells in 100 μL of binding medium and incubated at room temperature for 1 hr under gentle agitation. Unbound radioactivity was separated from cell-bound radioactivity by centrifugation through anoil gradient (80% silicon and 20% paraffin) on a microcentrifuge. Nonspecific binding was determined by a 200-fold molar excess of unlabeled CXCL8. Scatchard analysis was performed with the LIGAND program.

Cell lines

Human polymorphonuclear cells (PMN) and monocytes and rodent peritoneal PMN.

Preparation method

Soluble in DMSO. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

45 min (human PMN), 1 h (rodent PMN), or 2 h (monocytes).

Applications

Repertaxin inhibits human PMN migration induced by CXCL8 and CXCL1 with IC50 values of 1 nM and 400 nM respectively, which are mediated by CXCR1 and CXCR2, respectively. Repertaxin also inhibits rodent PMN chemotaxis induced by CXCL1 and CXCL2.

Animal models

Rat model of liver postischaemia RI.

Dosage form

3, 15, or 30 mg/kg; 15 min before reperfusion (i.v.) and 2 h after reperfusion (s.c.).

Applications

Repertaxin (15 mg/kg) inhibits PMN recruitment into reperfused livers by 90% and significantly reducesliver damage.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Bertini R, Allegretti M, Bizzarri C, et al. Noncompetitive allosteric inhibitors of the inflammatory chemokine receptors CXCR1 and CXCR2: prevention of reperfusion injury. Proc Natl Acad Sci U S A, 2004, 101(32): 11791-11796.