Development of radiopharmaceuticals containing anti-TNF monoclonal antibodies labeled with technetium-99m for use in the diagnosis of rheumatoid arthritis

Project no. 32087 · Status: Kapanmış (Yayın bekleniyor) · 2025

Summary

<p>Rheumatoid&nbsp;arthritis&nbsp;(RA)&nbsp;is&nbsp;a&nbsp;chronic&nbsp;inflammatory&nbsp;disease&nbsp;that&nbsp;requires&nbsp;early&nbsp;diagnosis&nbsp;and&nbsp;effective&nbsp;treatment.&nbsp;The&nbsp;development&nbsp;of&nbsp;targeted&nbsp;radiopharmaceuticals&nbsp;enabling&nbsp;molecular&nbsp;imaging&nbsp;of&nbsp;inflammation&nbsp;has&nbsp;become&nbsp;an&nbsp;important&nbsp;area&nbsp;of&nbsp;research.&nbsp;The&nbsp;aim&nbsp;of&nbsp;this&nbsp;study&nbsp;was&nbsp;to&nbsp;develop&nbsp;technetium-99m&nbsp;([^99mTc]Tc)-labeled&nbsp;radiopharmaceuticals&nbsp;based&nbsp;on&nbsp;adalimumab&nbsp;and&nbsp;golimumab&nbsp;for&nbsp;the&nbsp;diagnosis&nbsp;of&nbsp;RA&nbsp;and&nbsp;to&nbsp;compare&nbsp;the&nbsp;radiolabeling&nbsp;performance&nbsp;of&nbsp;different&nbsp;bifunctional&nbsp;chelators.</p><p>Adalimumab&nbsp;and&nbsp;golimumab&nbsp;monoclonal&nbsp;antibodies&nbsp;were&nbsp;conjugated&nbsp;with&nbsp;p-SCN-Bn-DTPA,&nbsp;p-SCN-Bn-DOTA,&nbsp;and&nbsp;S-acetyl-NHS-MAG3&nbsp;at&nbsp;different&nbsp;antibody-to-chelator&nbsp;molar&nbsp;ratios.&nbsp;The&nbsp;resulting&nbsp;conjugates&nbsp;were&nbsp;radiolabeled&nbsp;with&nbsp;[^99mTc]Tc,&nbsp;and&nbsp;radiochemical&nbsp;quality&nbsp;control&nbsp;was&nbsp;performed&nbsp;using&nbsp;instant&nbsp;thin-layer&nbsp;chromatography&nbsp;(ITLC).&nbsp;Different&nbsp;amounts&nbsp;of&nbsp;SnCl₂·2H₂O&nbsp;were&nbsp;evaluated&nbsp;during&nbsp;radiolabeling&nbsp;to&nbsp;determine&nbsp;the&nbsp;optimum&nbsp;labeling&nbsp;conditions.&nbsp;The&nbsp;radiochemical&nbsp;stability&nbsp;of&nbsp;the&nbsp;selected&nbsp;formulations&nbsp;was&nbsp;investigated&nbsp;at&nbsp;room&nbsp;temperature&nbsp;and&nbsp;in&nbsp;different&nbsp;cell&nbsp;culture&nbsp;media.&nbsp;In&nbsp;addition,&nbsp;the&nbsp;<em>in&nbsp;vitro</em>&nbsp;cellular&nbsp;uptake&nbsp;of&nbsp;the&nbsp;radiopharmaceuticals&nbsp;was&nbsp;evaluated&nbsp;using&nbsp;L929&nbsp;mouse&nbsp;fibroblast,&nbsp;RAW264.7&nbsp;mouse&nbsp;macrophage,&nbsp;and&nbsp;SW982&nbsp;human&nbsp;synovial&nbsp;cell&nbsp;lines.</p><p>Among&nbsp;the&nbsp;investigated&nbsp;bifunctional&nbsp;chelators,&nbsp;MAG3-based&nbsp;formulations&nbsp;exhibited&nbsp;the&nbsp;highest&nbsp;radiochemical&nbsp;purity&nbsp;for&nbsp;both&nbsp;monoclonal&nbsp;antibodies.&nbsp;The&nbsp;highest&nbsp;radiochemical&nbsp;purity&nbsp;was&nbsp;obtained&nbsp;with&nbsp;MAG3&nbsp;conjugates&nbsp;prepared&nbsp;at&nbsp;an&nbsp;antibody-to-chelator&nbsp;molar&nbsp;ratio&nbsp;of&nbsp;1:100&nbsp;and&nbsp;radiolabeled&nbsp;using&nbsp;100&nbsp;µg&nbsp;SnCl₂·2H₂O,&nbsp;reaching&nbsp;94.2%&nbsp;for&nbsp;adalimumab&nbsp;and&nbsp;93.5%&nbsp;for&nbsp;golimumab.&nbsp;The&nbsp;optimum&nbsp;formulations&nbsp;maintained&nbsp;radiochemical&nbsp;purity&nbsp;above&nbsp;90%&nbsp;for&nbsp;up&nbsp;to&nbsp;6&nbsp;hours&nbsp;at&nbsp;room&nbsp;temperature&nbsp;and&nbsp;for&nbsp;2&nbsp;hours&nbsp;in&nbsp;different&nbsp;cell&nbsp;culture&nbsp;media.&nbsp;Cellular&nbsp;uptake&nbsp;studies&nbsp;demonstrated&nbsp;the&nbsp;highest&nbsp;uptake&nbsp;in&nbsp;SW982&nbsp;cells,&nbsp;followed&nbsp;by&nbsp;RAW264.7&nbsp;cells,&nbsp;while&nbsp;the&nbsp;lowest&nbsp;uptake&nbsp;was&nbsp;observed&nbsp;in&nbsp;L929&nbsp;cells.&nbsp;Furthermore,&nbsp;[^99mTc]Tc-MAG3-adalimumab&nbsp;exhibited&nbsp;higher&nbsp;cellular&nbsp;uptake&nbsp;than&nbsp;[^99mTc]Tc-MAG3-golimumab&nbsp;in&nbsp;all&nbsp;tested&nbsp;cell&nbsp;lines.</p><p>In&nbsp;conclusion,&nbsp;MAG3-conjugated&nbsp;adalimumab&nbsp;and&nbsp;golimumab&nbsp;were&nbsp;successfully&nbsp;radiolabeled&nbsp;with&nbsp;[^99mTc]Tc,&nbsp;demonstrating&nbsp;high&nbsp;radiochemical&nbsp;purity,&nbsp;satisfactory&nbsp;radiochemical&nbsp;stability,&nbsp;and&nbsp;favorable&nbsp;<em>in&nbsp;vitro</em>&nbsp;cellular&nbsp;uptake&nbsp;characteristics.&nbsp;These&nbsp;findings&nbsp;indicate&nbsp;that&nbsp;the&nbsp;developed&nbsp;radiopharmaceuticals&nbsp;are&nbsp;promising&nbsp;candidates&nbsp;for&nbsp;the&nbsp;molecular&nbsp;imaging&nbsp;of&nbsp;rheumatoid&nbsp;arthritis.</p><p><strong>Keywords</strong>;&nbsp;rheumatoid&nbsp;arthritis,&nbsp;technetium-99m,&nbsp;adalimumab,&nbsp;golimumab,&nbsp;MAG3,&nbsp;radiopharmaceutical,&nbsp;molecular&nbsp;imaging.</p>

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