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Further investigations of morpholino pretargeting in mice—establishing quantitative relations in tumor

  • Molecular Imaging
  • Published:
European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

This laboratory has previously published on phosphorodiamidate morpholino (MORF) pretargeting of tumor in which an anti-tumor antibody conjugated with MORF (a DNA analogue) is first administered, followed at a later time by the radiolabeled complementary MORF (cMORF) as the effector. In the present study, the pharmacokinetics of the antibody and effector were measured under different conditions in mice to establish their quantitative relationships with tumor accumulations by pretargeting.

Methods

A cytosine-free 18 mer cMORF was conjugated with MAG3 for 99mTc labeling while the anti-CEA antibody MN14 was conjugated with DTPA for 111In labeling and with MORF to impart binding affinity for radiolabeled cMORF. Mice bearing LS174T thigh tumors were used to study: (1) the pharmacokinetics of MN14-MORF by administering 111In-MN14 at doses between 10 and 100 μg with sacrifice at 2 days and at 30 μg with sacrifice between 1 and 3 days; (2) the biodistribution of 99mTc-cMORF following one to four injections (containing 0.15 μg each and separated by 1 h) to animals having received 30 μg of antibody–MORF 2 days earlier and with sacrifice at 3 h after the final injection; and (3) the influence on the biodistribution of 99mTc-cMORF of a 2 to 4 day interval between the administration of 30 μg of antibody–MORF and 0.30 μg of 99mTc-cMORF.

Results

(1) The biodistribution of antibody in percent accumulation (%ID or %ID/g) was largely independent of antibody dose but the absolute accumulation of antibody in tumor increased linearly with dose, showing no evidence of tumor saturation of CEA sites by MN14. Over 1–3 days post antibody administration, blood levels of radiolabeled antibody decreased as expected; however, tumor levels remained constant, thus showing an absence of antibody clearance in tumor over this period. (2) With fixed antibody–MORF dose and increasing number of injections of 99mTc-cMORF, cumulative percent blood levels steadily decreased in agreement with the values calculated based on the antibody–MORF in blood. In contrast, cumulative percent tumor levels stayed fairly constant over the first two injections. Thus the antibody–MORF in tumor became saturated with cMORF more slowly than that in blood owing to delivery differences. (3) As expected, percent blood levels decreased with increasing interval between injections of antibody–MORF and 99mTc-cMORF. The percent tumor accumulation, however, remained constant over the 3 day interval, thus demonstrating only slow loss of MORF expression in situ. The 99mTc-cMORF accumulation in tumor after saturation was mathematically determined based on the antibody–MORF concentration in tumor while the blood levels of 99mTc-cMORF were determined based on the concentration of antibody-MORF in blood.

Conclusion

Contrary to conclusions arrived at in our earlier study, the results of this study show that tumor CEA sites were not saturated even at the highest antibody dose investigated, that accessibility of MORF sites in tumor by 99mTc-cMORF was unhindered and that the maximum percent tumor accumulation of 99mTc-cMORF depended only on the tumor delivery efficiency of 99mTc-cMORF.

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Acknowledgements

The authors are grateful to Dr. Gary L. Griffiths of Immunomedics (Morris Plains, NJ) for providing the MN14. We also thank Loretta Lee of the UMMS Tissue Culture Center for maintaining the LS174T tumor cells. Financial support for this investigation was provided in part by the National Institutes of Health (CA79507, CA94994, and CA107360).

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Authors and Affiliations

  1. Division of Nuclear Medicine, Department of Radiology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA, 01655-0243, USA

    Guozheng Liu, Jiang He, Shuping Dou, Suresh Gupta, Mary Rusckowski & Donald J. Hnatowich

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  1. Guozheng Liu
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  2. Jiang He
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  3. Shuping Dou
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  4. Suresh Gupta
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  6. Donald J. Hnatowich
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Correspondence to Guozheng Liu.

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Liu, G., He, J., Dou, S. et al. Further investigations of morpholino pretargeting in mice—establishing quantitative relations in tumor. Eur J Nucl Med Mol Imaging 32, 1115–1123 (2005). https://doi.org/10.1007/s00259-005-1853-5

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  • DOI: https://doi.org/10.1007/s00259-005-1853-5

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