title: Deferiprone (L1) chelates pathologic iron deposits from membranes of intact thalassemic and sickle red blood cells both in vitro and in vivo.
abstract: Red blood cell (RBC) membranes from patients with the thalassemic and sickle hemoglobinopathies carry abnormal deposits of iron presumed to mediate a variety of oxidative-induced membrane dysfunctions. We hypothesized that the oral iron chelator deferiprone (L1), which has an enhanced capacity to permeate cell membranes, might be useful in chelating these pathologic iron deposits from intact RBCs. We tested this hypothesis in vitro by incubating L1 with RBCs from 15 patients with thalassemia intermedia and 6 patients with sickle cell anemia. We found that removal of RBC membrane free iron by L1 increased both as a function of time of incubation and L1 concentration. Thus, increasing the time of incubation of thalassemic RBCs with 0.5 mmol/L L1 from 0.5 to 6 hours, enhanced removal of their membrane free iron from 18% +/- 9% to 96% +/- 4%. Dose-response studies showed that incubating thalassemic RBC for 2 hours with L1 concentrations ranging from 0.125 to 0.5 mmol/L resulted in removal of membrane free iron from 28% +/- 15% to 68% +/- 11%. Parallel studies with sickle RBCs showed a similar pattern in time and dose responses. Deferoxamine (DFO), on the other hand, was ineffective in chelating membrane free iron from either thalassemic or sickle RBCs regardless of dose (maximum, 0.333 mmol/L) or time of incubation (maximum, 24 hours). In vivo efficacy of L1 was shown in six thalassemic patients whose RBC membrane free iron decreased by 50% +/- 29% following a 2-week course of L1 at a daily dose of 25 mg/kg. As the dose of L1 was increased to 50 mg/kg/d (n = 5), and then to 75 mg/kg/d (n = 4), 67% +/- 14% and 79% +/- 11%, respectively, of their RBC membrane free iron was removed. L1 therapy--both in vitro and in vivo--also significantly attenuated the malondialdehyde response of thalassemic RBC membranes to in vitro stimulation with peroxide. Remarkably, the heme content of RBC membranes from L1-treated thalassemic patients decreased by 28% +/- 10% during the 3-month study period. These results indicate that L1 can remove pathologic deposits of chelatable iron from thalassemic and sickle RBC membranes, a therapeutic potential not shared by DFO. Furthermore, membrane defects possibly mediated by catalytic iron, such as lipid peroxidation and hemichrome formation, may also be alleviated, at least in part, by L1.
authors.affiliation: Department of Medicine, Hadassah-Hebrew University Hospital, Jerusalem, Israel.
authors.forename: O
authors.initials: O
authors.lastname: Shalev
authors.forename: T
authors.initials: T
authors.lastname: Repka
authors.forename: A
authors.initials: A
authors.lastname: Goldfarb
authors.forename: L
authors.initials: L
authors.lastname: Grinberg
authors.forename: A
authors.initials: A
authors.lastname: Abrahamov
authors.forename: N F
authors.initials: NF
authors.lastname: Olivieri
authors.forename: E A
authors.initials: EA
authors.lastname: Rachmilewitz
authors.forename: R P
authors.initials: RP
authors.lastname: Hebbel
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood*; D000077543:Deferiprone; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007501:Iron / Q000097:blood*; D007502:Iron Chelating Agents / Q000494:pharmacology / Q000627:therapeutic use; D015227:Lipid Peroxidation / Q000187:drug effects; D008315:Malondialdehyde / Q000097:blood; D011728:Pyridones / Q000494:pharmacology* / Q000627:therapeutic use*; D013156:Splenectomy; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D017086:beta-Thalassemia / Q000097:blood* / Q000628:therapy
chemical_list: D007502:Iron Chelating Agents; D011728:Pyridones; D017392:Thiobarbituric Acid Reactive Substances; D000077543:Deferiprone; D008315:Malondialdehyde; D007501:Iron