Delivery of radionuclides to pretargeted monoclonal antibodies using dihydrofolate reductase and methotrexate in an affinity system.
A novel affinity system for a two-phase delivery of radionuclides to tumor cells has been developed. In the first phase, a nontoxic bivalent monoclonal antibody conjugated to an enzyme is targeted to the tumor cells. In the second phase, a radionuclide-derivatized enzyme inhibitor, specific for the enzyme conjugated to the antibody, is administered. The model system selected for this study is the recombinant human enzyme dihydrofolate reductase (rhDHFR) and its high-affinity competitive inhibitor methotrexate (MTX). MTX was labeled with a radionuclide by covalent attachment of diethylenetriaminepentaacetic acid (DTPA) complexed with 111In. Using the gamma-carboxyl residue of MTX for the attachment of DTPA, binding of the inhibitor to rhDHFR was not affected. The inhibitory activities of nonderivatized MTX and DTPA-MTX were indistinguishable. Human K562 erythroleukemia cells were used to evaluate under in vitro conditions the DHFR-MTX affinity system for the delivery of 111In-labeled DTPA-MTX to pretargeted alpha-transferrin receptor antibody-rhDHFR conjugates (alpha-TFR-DHFR). The data demonstrate that the delivery of 111In is dose dependent and highly specific. Under saturating conditions, binding of 111In-DTPA-MTX to alpha-TFR-DHFR-treated cells was 14-fold higher than to cells treated with nonconjugated alpha-TFR antibody. Further experiments indicated that the low level of nonspecific binding of 111In-DTPA-MTX was comparable to that of 111In-DTPA, known for its complete extracellular distribution and rapid clearance through the kidneys. Based on the data of this study, antibody-conjugated rhDHFR and radionuclide-labeled DTPA-MTX complexes provide components for an alternative radioimmunotherapeutic approach that can be expected to result in improved tumor tissue ratios of both the targeting moiety and the radionuclide-labeled derivative as compared to current approaches.
Hawkins
G A
GA
Organon Teknika Biotechnology Research Institute, Rockville, Maryland 20850.
McCabe
R P
RP
Kim
C H
CH
Subramanian
R
R
Bredehorst
R
R
McCullers
G A
GA
Vogel
C W
CW
Hanna
M G
MG
Pomato
N
N
D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000627:therapeutic use; D004337:Drug Carriers; D005493:Folic Acid Antagonists; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D000922:Immunotoxins / Q000378:metabolism* / Q000627:therapeutic use; D007205:Indium Radioisotopes / Q000008:administration & dosage*; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism / Q000532:radiotherapy; D008727:Methotrexate / Q000378:metabolism / Q000493:pharmacokinetics*; D004369:Pentetic Acid / Q000378:metabolism / Q000493:pharmacokinetics*; D016499:Radioimmunotherapy; D011990:Receptors, Transferrin / Q000276:immunology; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D004337:Drug Carriers; D005493:Folic Acid Antagonists; D007074:Immunoglobulin G; D000922:Immunotoxins; D007205:Indium Radioisotopes; D011990:Receptors, Transferrin; D004369:Pentetic Acid; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate