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Prednisolone has been shown to be teratogenic in mice when given in doses 1-10 times the human ocular dose. Dexamethasone, hydrocortisone and prednisolone were applied to both eyes of pregnant mice five times per day on days 10 through 13 of gestation. Transgenic mice that constitutively express BAFF develop an SLE‐like syndrome associated with autoantibodies against nuclear antigens (4, 5). Clinical studies have demonstrated that BAFF is overexpressed, either intermittently or persistently, in many patients with SLE ( 6, 7 ), and the levels have correlated with serologic and clinical. In regard to our ENT1 model, there are no significant compensatory changes noted in gene expression of nucleoside transporter or metabolism after genetic deletion of murine ENT1. 34, 35 Likewise, previous studies have indicated that genetic deficiency of OATP1B2 is not associated with any pronounced compensatory alterations in metabolic enzyme.

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Abstract

Purpose: Nitrobenzylthioinosine (NBMPR), a potent inhibitor of nucleoside transport in many cell types, modulates the in vivo disposition of several cytotoxic nucleoside analogs. In this study, a radioligand binding assay was developed for measurement of the NBMPR content of plasma and erythrocytes. Methods: The assay was based on the competition between NBMPR and [3H]NBMPR for high-affinity sites on human erythrocyte membranes. With this assay, we followed in mice changes in the NBMPR content of blood plasma and erythrocytes, following the intraperitoneal injection of the disodium salt of NBMPR 5′-monophosphate (NBMPR-P), a prodrug form of NBMPR. Results: The radioligand binding assay was able to measure precisely as little as 2.5 pmol of NBMPR, allowing the direct determination of NBMPR concentrations in plasma as low as 16 nM. As few as 8 × 103 molecules of NBMPR per cell could be determined in erythrocytes. The NBMPR content of plasma from mice injected with NBMPR-P was maximal at about 20 min after injection and declined to <0.2% of the peak value by 10 h. Erythrocyte-associated NBMPR was also maximal at 20 min, and declined to 11% of the peak value by 10 h after injection. Time courses for the disappearance of NBMPR from plasma and erythrocytes were monoexponential and yielded half-life values of 0.39 h and 0.68 h, respectively, an apparent volume of distribution of 0.61 l/kg, and a clearance of 1.1 l/h per kg. Conclusions: The radioligand binding assay is a sensitive and facile method for monitoring NBMPR concentrations in mammalian plasma and tissue extracts.

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