Radioimmunoassay for mitoxantrone, a new antitumor agent

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Abstract
A sensitive and specific radioimmunoassay for mitoxantrone in serum has been developed. The procedure allows direct measurement of mitoxantrone in unextracted serum samples, by using antisera from rabbits immunized with mitoxantrone-BSA antigen. Tritiated mitoxantrone of high specific radioactivity (ca. 15 Ci/mmol) was used as a radio-tracer ligand. The assay allows the detection of as little as 50 pg/ml and the quantitation of 75 pg/ml in 0.5 ml serum samples. Standard curves were linear in the concentration range of 75–2500 pg/ml, at antiserum dilutions of 1:15,000. The assay shows good reproducibility: coefficients of variation of 3–6% were obtained by analyzing five samples/concentration at 75, 100, 250, 500, and 1000 pg/ml. There was no cross reactivity with the major metabolite in human serum, having concentrations of up to 10,000 pg/ml. Serum samples collected at various time intervals from rats dosed intravenously with mitoxantrone (0.5 mg/kg), were analyzed for unchanged mitoxantrone by RIA. The drug concentrations decreased from 32 ng/ml at 0.5 h to 0.45 ng/ml by 24 h after dosing. Mitoxantrone Mitoxantrone is being currently produced by American Cyanamid — Lederle laboratories under the trademark NOVANTRONETM. , a dihydroxyanthracenedione derivative (1), is an antitumor agent currently used in clinical trials with very encouraging results, especially in metastatic breast cancer, and low incidence of adverse reactions (2–4). The drug is being administered intravenously at doses up to 14 mg/m2. Preliminary pharmacokinetic studies (to be published separately) indicate rapid distribution, followed by slow clearance rates from the tissues. Mitoxantrone is being currently produced by American Cyanamid — Lederle laboratories under the trademark NOVANTRONETM. The sensitivity of the currently available (HPLC) methods (5, 6) is of about 5–20 ng/ml in serum. The purpose of this work was to develop a more sensitive method, such as radioimmunoassay, which can be utilized in pharmacokinetic studies to measure mitoxantrone levels in biological fluids, over extended periods of time after dosing.