Complementation Dependent Enzyme Prodrug Therapy Enables Targeted Activation of Prodrug on HER2-Positive Cancer Cells

• Published in: ACS medicinal chemistry letters Vol. 13; no. 11; pp. 1769 - 1775
• Main Authors: Nervig, Christine S., Hatch, Samuel T., Owen, Shawn C.
• Format: Journal Article
• Language: English
• Published: American Chemical Society 10.11.2022
• Subjects: Letter; prodrug; targeted delivery; Antibody-directed enzyme prodrug therapy; HER2; complementation; split enzyme
• ISSN: 1948-5875; 1948-5875
• DOI: 10.1021/acsmedchemlett.2c00394

Antibodies have been explored for decades for the delivery of small molecule cytotoxins directly to diseased cells. In antibody-directed enzyme prodrug therapy (ADEPT), antibodies are armed with enzymes that activate nontoxic prodrugs at tumor sites. However, this strategy failed clinically due to off-target toxicity associated with the enzyme prematurely activating prodrug systemically. We describe here the design of an antibody-fragment split enzyme platform that regains activity after binding to HER2, allowing for site-specific activation of a small molecule prodrug. We evaluated a library of fusion constructs for efficient targeting and complementation to identify the most promising split enzyme pair. The optimal pair was screened for substrate specificity among chromogenic, fluorogenic, and prodrug substrates. Evaluation of this system on HER2-positive cells revealed 7-fold higher toxicity of the activated prodrug over prodrug treatment alone. Demonstrating the potential of this strategy against a known clinical target provides the basis for a unique therapeutic platform in oncology.