Pharmacokinetics and Catabolism of [3H]TAK-164, a Guanylyl Cyclase C Targeted Antibody-Drug Conjugate.

Abstract

TAK-164 is an antibody-drug conjugate (ADC) comprising human anti-guanylyl cyclase C (GCC) monoclonal antibody conjugated to indolinobenzodiazepine DNA alkylator IGN-P1 through a cleavable alanine-alanine dipeptide linker. TAK-164 is currently being evaluated for the treatment of gastrointestinal cancers expressing GCC. The catabolism of TAK-164 was studied using 3H-labeled ADC using GCC-expressing HEK-293 (GCC-HEK-293) cells, rat tritosomes, cathepsin B, and tumor-bearing mice. Time- and target-dependent uptake of [3H]TAK-164 was observed in GCC-HEK-293 cells with approximately 12% of radioactivity associated with DNA after 24 hours of incubation. Rat liver tritosomes and cathepsin B yielded IGN-P1 aniline, sulfonated IGN-P1 (s-IGN-P1) aniline, and a lysine conjugate of IGN-P1 (IGN-P1-Lys) aniline as catabolites. In tumor-bearing mice, [3H]TAK-164 exhibited a terminal half-life of approximately 41 and 51 hours in plasma and blood, respectively, with low plasma clearance (0.75 ml/h per kilogram). The extractable radioactivity in plasma and tumor samples revealed the presence of s-IGN-P1 aniline and IGN-P1 aniline as payload-related components. The use of a radiolabeled payload in the ADC in tumor uptake investigations provided direct and quantitative evidence for tumor uptake, DNA binding, and proof of mechanism of action of the payload. SIGNIFICANCE STATEMENT: Since payload-related species are potent cytotoxins, a thorough characterization of released products of ADCs, metabolites, and their drug interaction potential is necessary prior to clinical investigations. This study characterized in vitro and in vivo DNA binding mechanisms and released products of TAK-164. The methodologies described here will be highly useful for characterization of payload-related products of ADCs in general.

• Authors: Jayaprakasam Bolleddula, Abhi Shah, Mohammad Shadid, Afrand Kamali, Michael D Smith, Swapan K Chowdhury
• Journal: Drug metabolism and disposition: the biological fate of chemicals (Drug Metab Dispos) Vol. 48 Issue 11 Pg. 1239-1245 (11 2020) ISSN: 1521-009X [Electronic] United States
• PMID: 32843329 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 by The American Society for Pharmacology and Experimental Therapeutics.

Chemical References

• Antineoplastic Agents
• Immunoconjugates
• Recombinant Proteins
• CTSB protein, human
• Cathepsin B
• GUCY2C protein, human
• Receptors, Enterotoxin

Topics

• Animals
• Antineoplastic Agents (administration & dosage, pharmacokinetics)
• Cathepsin B (metabolism)
• Cell Line, Tumor
• Female
• HEK293 Cells
• Half-Life
• Humans
• Immunoconjugates (administration & dosage, pharmacokinetics)
• Microsomes, Liver
• Neoplasms (drug therapy, pathology)
• Rats
• Receptors, Enterotoxin (antagonists & inhibitors, metabolism)
• Recombinant Proteins (metabolism)
• Xenograft Model Antitumor Assays