| Abstract: |
Objective: Leukocyte function-associated antigen-1 (LFA-1) is constitutively expressed on leukocytes, including overexpression on lymphomas and leukemias. We have developed a derivative of BIRT 377, an allosteric inhibitor of LFA-1, which may be chemically tagged without affecting binding. In this study, we modified this derivative, (R)-1-(4-aminobutyl)-5-(4-bromobenzyl) -3-(3,5-dichlorophenyl)-5-methylimidazolidine-2,4-dione (butylamino-NorBIRT), and demonstrated its potential as a noninvasive imaging agent. Methods: Specific binding of fluorescein-labeled butylamino-NorBIRT to both human and murine cells was demonstrated using equilibrium binding and dissociation techniques. A radiometal, lutetium-177 (Lu-177), was incorporated into the butylamino-NorBIRT through 1,4,7,10-tetraazacyclododecane-N,N′,N″,N‴-tetraacetic acid (DOTA) as a chelator. Results: Equilibrium-binding experiments demonstrated that fluorescein-labeled butylamino-NorBIRT specifically binds human and murine LFA-1 with affinity constants of 135 and 186 nM, respectively. Dissociation kinetic experiments demonstrated an off-rate of 0.168/second -1 on murine cells, consistent with the observed affinity constant. Lutetium-177 was used for labeling, with ≥99.99% radiochemical purity and incorporation yield. This radiolabeled derivative exhibited high stability in fetal bovine serum (FBS) at 37°C over 72 hours. 177Lu-DOTA- butylamino-NorBIRT showed a binding affinity of 235 nM to human LFA-1 for equilibrium binding and competitive binding experiments. Conclusions: The radiolabeled DOTA-butylamino-NorBIRT may have potential as a noninvasive imaging or therapeutic agent in both human and mouse models. © Mary Ann Liebert, Inc. |
| Keywords: |
controlled study; leukemia; unclassified drug; human cell; nonhuman; binding affinity; antigen expression; animal cell; mouse; animals; mice; gene overexpression; animal experiment; animal model; kinetics; isotope labeling; lymphoma; molecular structure; radioisotopes; non invasive measurement; leukocyte; organometallic compounds; drug binding; radionuclide imaging; binding competition; radiochemistry; lymphocyte function-associated antigen-1; allosterism; dissociation constant; molecular stability; binding, competitive; heterocyclic compounds, 1-ring; lutetium; lymphocyte function associated antigen 1; hl-60 cells; equilibrium constant; lutetium 177; butylamino-norbirt; leukocyte function-associated antigen-1; lutetium-177; 1 (4 aminobutyl) 5 (4 bromobenzyl) 3 (3,5 dichlorophenyl) 5 methylimidazolidine 2,4 dione; 1,4,7,10 tetraazacyclododecane n,n',n'',n''' tetraacetic acid butylamino 1 (4 aminobutyl) 5 (4 bromobenzyl) 3 (3,5 dichlorophenyl) 5 methylimidazolidine 2,4 dione; 1,4,7,10 tetraazacyclododecane n,n',n'',n''',tetraacetic acid; birt 377; lymphocyte function associated antigen 1 derivative; hydantoins; imidazolidines
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