A Naphthoquinone Derivative Can Induce Anemia through Phosphatidylserine Exposure-Mediated Erythrophagocytosis

Abstract

A naphthoquinone derivative, ␤-lapachone (␤L; 3,4-dihydro-2,2-dimethyl-2H-naphthol[1,2-b]pyran-5,6-dione), is receiving huge attention for its potent therapeutic effects against various diseases. However, during the preclinical safety evaluation, repeated oral treatment of ␤L in rats induced anemia, i.e., a significantly decreased erythrocyte count. In this study, in an effort to elucidate the mechanism underlying the ␤L-induced anemia, we investigated the effects of ␤L on erythrocytes with freshly isolated human erythrocytes in vitro and rat in vivo. ␤L did not induce erythrocyte hemolysis, indicating that direct hemotoxicity was not involved in ␤L-associated anemia. Meanwhile, phosphatidylserine (PS) exposure along with spherocytic shape change and microvesicle generation, important factors in the facilitation of erythrophagocytosis, were increased significantly by ␤L. The PS exposure on erythrocytes was from ␤L-induced reactive oxygen species generation and subsequent depletion of reduced glutathione and protein thiol, which culminated in the modified activities of phospholipid translocases, i.e., inhibition of flippase and activation of scramblase. It is important to note that coincubation of macrophage with ␤L-treated erythrocyte in vitro showed increased erythrophagocytosis, demonstrating that the removal of erythrocyte by macrophage can be facilitated by ␤L-induced PS exposure. In good accordance with these in vitro results, after oral administration of ␤L in rats, increased PS exposure and depletion of glutathione were observed along with enhanced splenic sequestration of erythrocytes. In conclusion, these results suggest that ␤L-induced anemia might be mediated through the PS exposure and subsequent erythrophagocytosis, providing novel insight into the drug-induced anemia.

An NQO1-Initiated and p53-Independent Apoptotic Pathway Determines the Anti-Tumor Effect of Tanshinone IIA against Non-Small Cell Lung Cancer

“…β-Lapachone (Lap), a well studied NQO1 substrate, has been identified as a promising agent for various cancer therapy [18]. However, repeated oral treatment of Lap induces anemia in both rats and humans which may greatly limit its application [19], [20]. Another limitation of Lap is its NQO1 specific activity can only be realized within a relatively narrow concentration range and exposure time window [18]; it is questionable whether such a condition can be well controlled in the in vivo states.…”

β-Lapachone induces heart morphogenetic and functional defects by promoting the death of erythrocytes and the endocardium in zebrafish embryos

“…A recent study also showed that oral treatment of rats with β-lapachone induced anemia that was attributed to increased phosphatidylserine externalization and erythrophagocytosis induced by ROS [56]. However, the extents of reduction in erythrocyte cell numbers in adult rats (22%~26%) and zebrafish embryos (75%~90%) upon β-lapachone treatment greatly differed.…”

Therapeutic potential of manipulating suicidal erythrocyte death

“…Nevertheless, any of the small molecules listed in Table 2 could, at least in theory, accelerate the eryptosis of plasmodium-infected erythrocytes and thus counteract the rise of parasitemia. It is noteworthy, that the malaria pathogen plasmodium is not expected to become resistant to therapeutic acceleration of eryptosis, which is accomplished by cell mechanisms and is Estramustine 75 --100 µM + + [67] Ferutinin 30 µM + [68] Fluoxetine 25 --50 µM + [69] FTY720 10 µM + [1,2] Fumagillin 10 --100 µM + + [70] Gambogic acid 100 --500 nM + + [1,2] Gedunin 12 µM + [71] Geldanamycin 5 --50 µM + + [72] Glycation 40 mM glucose + Honokiol 5 --15 µM + + [74] Indoxyl sulfate 50 --600 µM + + Lumefantrine 3 --24 µg/ml [76] Lysophosphatidic acid 2.5 µM + + [77] Mercury 1 µM + + [78] Methyldopa 6 µg/ml + + Mitoxantrone 10 µg/ml + + [55] Monensin 0.1 µg/ml + [1,2] Mushroom tyrosinase 7 U/mL + + [81] Naphthoquinone derivatives 10 µM + [82] Nitazoxanide 1 --50 µg/ml + [83] Novobiocin 500 µM + + [84] Nystatin 5 --15 µg/ml + [85] Ochratoxin A 2.5 --10 µM + + Patulin 2.5 --10 µM + [87] Penta-O-galloyl-b-D-glucose 10 --50 µM + + [88] Peptidoglycan 10 µg/ml + [89] Phloretin 100 µM + [90] Phorbol-12 myristate-13 acetate…”