²¹¹At-AntiCD33 in NMRI nu/nu mice

Summary

The aim of this study is to verify the in vivo stability, to determine the biodistribution and to estimate the unspecific radiotoxicity of an ²¹¹At-labelled CD33-antibody (²¹¹At-anti-CD33) in mice with a view to therapeutic application in treating leukaemia. Animals, methods: ²¹¹At was produced via the ²⁰⁹Bi(α,2n)²¹¹At reaction and was linked via 3-²¹¹At-succinimidyl-benzoate to the anti-CD33-antibody. The biodistribution and the in vivo stability in serum were determined after i.v.-injection in NMRI nu/nu-mice. For toxicity experiments, mice received either three times 315–650 kBq ²¹¹At-antiCD33 or unlabelled antibody and NaCl-solution respectively. Results: ²¹¹At-antiCD33 showed a characteristic biodistribution complying with the unspecific antibody retention in the reticular endothelial system. The largest proportion of radioactivity remained in blood and blood-rich tissues with a minor accumulation in the thyroid and stomach. After 21 h, > 85% of activity in serum still represented intact antibody. Mice showed no difference in unspecific toxicity of ²¹¹At-labelled antibodies over six months compared to those treated with unlabelled antibody and NaCl-solution respectively, with regard to histopathologic lesions, survival time, behaviour and haemograms. Conclusion: The radiolabelling method yielded adequate in vivo stability of ²¹¹At-antiCD33. Biodistribution with rapid elimination of free ²¹¹At via kidneys and urine complies with requirements for targeted therapy. Activity doses potentially required for treatment do not elicit radiotoxicity to normal organs in mice. Further development is required to enhance the apparent specific activity and to verify the efficacy in an adequate animal model before phase I clinical studies in leukaemia can be envisaged.

Zusammenfassung

Ziel der Studie war es, die in vivo Stabilität und die Biodistribution eines ²¹¹At-markierten Anti-CD33-Antikörpers (²¹¹At-AntiCD33) zu untersuchen und dessen unspezifische Radiotoxizität hinsichtlich des Einsatzes in der Leukämietherapie anhand eines Mausmodells abzuschätzen. Tiere, Methoden: ²¹¹At wurde über die ²⁰⁹Bi(α,2n)²¹¹At-Reaktion am Zyklotron erzeugt und als Succinimidyl-3-²¹¹At-benzoat an den Antikörper gebunden. Die Biodistribution und die In-vivo-Stabilität im Serum wurden nach intravenöser Injektion an NMRI-nu/ nu-Mäusen evaluiert. Für die Experimente zur Radiotoxizität wurde den Mäusen dreimal 315–650 kBq ²¹¹At-Anti-CD33 bzw. unmarkierter Antikörper oder NaCl-Lösung verabreicht. Ergebnisse: ²¹¹At-Anti-CD33 zeigte eine charakteristische Bioverteilung mit einer unspezifischen Retention des Antikörpers im retikuloendothelialen System. Der größte Anteil der Radioaktivität verblieb im Blut und in blutreichen Geweben, eine schwächere Anreicherung wurde in Schilddrüse und Magen beobachtet. Nach 21 h betrug der Anteil an intakten ²¹¹At-Anti-CD33 > 85% der Serumaktivität. Zwischen den Behandlungsgruppen war histopathologisch, in der Überlebenszeit und im Blutbild über sechs Monate kein Unterschied in der unspezifischen Toxizität des ²¹¹At-Anti-CD33 zu erkennen. Schlussfolgerung: Die Synthesemethode führt zu einem in vivo ausreichend stabilen Produkt. Die Biodistribution mit schneller renaler Ausscheidung ist für eine therapeutische Anwendung geeignet. Eine für therapeutische Zwecke angemessene Dosierung führte zu keinen signifikanten radiotoxischen Organschäden im Mausmodell. Weitere Untersuchungen zur Verbesserung der scheinbaren spezifischen Aktivität und zur Wirksamkeit sind nötig, bevor eine klinische Studie (Phase I) an Leukämie-Patienten geplant werden kann.

Keywords

Astatine - radioimmunotherapy - leukaemia - CD33 antigen - experimental animal model

Schlüsselwörter

Astat - Radioimmunotherapie - Leukämie - CD33-Antigen - tierexperimentelles Modell

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