Chemical Site-Specific Conjugation Platform to Improve the Pharmacokinetics and Therapeutic Index of Antibody–Drug Conjugates

• Published in: Molecular pharmaceutics 2021-11, Vol.18 (11), p.4058-4066
• Main Authors: Matsuda, Yutaka, Seki, Takuya, Yamada, Kei, Ooba, Yuri, Takahashi, Kazutoshi, Fujii, Tomohiro, Kawaguchi, Sayaka, Narita, Takahiro, Nakayama, Akira, Kitahara, Yoshiro, Mendelsohn, Brian A, Okuzumi, Tatsuya
• Format: Article
• Language: English
• Subjects: Ado-Trastuzumab Emtansine - administration & dosage; Ado-Trastuzumab Emtansine - pharmacokinetics; Animals; Antineoplastic Agents - administration & dosage; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacokinetics; Antineoplastic Agents - toxicity; Chemistry, Pharmaceutical; Drug Compounding - methods; Drug Stability; Female; Humans; Immunoconjugates - administration & dosage; Immunoconjugates - chemistry; Immunoconjugates - pharmacokinetics; Immunoconjugates - toxicity; Maximum Tolerated Dose; Mice; Neoplasms - drug therapy; Neoplasms - pathology; Rats; Therapeutic Index; Toxicity Tests, Acute; Xenograft Model Antitumor Assays
• ISSN: 1543-8384; 1543-8392
• DOI: 10.1021/acs.molpharmaceut.1c00473

To overcome a lack of selectivity during the chemical modification of native non-engineered antibodies, we have developed a technology platform termed “AJICAP” for the site-specific chemical conjugation of antibodies through the use of a class of IgG Fc-affinity reagents. To date, a limited number of antibody–drug conjugates (ADCs) have been synthesized via this approach, and no toxicological study was reported. Herein, we describe the compatibility and robustness of AJICAP technology, which enabled the synthesis of a wide variety of ADCs. A stability assessment of a thiol-modified antibody synthesized by AJICAP technology indicated no appreciable increase in aggregation or decomposition upon prolonged storage, indicating that the unexpectedly stable thiol intermediate has a great potential intermediate for payload or linker screening or large-scale manufacturing. Payload conjugation with this stable thiol intermediate generated several AJICAP-ADCs. In vivo xenograft studies indicated that the AJICAP-ADCs displayed significant tumor inhibition comparable to benchmark ADC Kadcyla. Furthermore, a rat pharmacokinetic analysis and toxicology study indicated an increase in the maximum tolerated dose, demonstrating an expansion of the AJICAP-ADC therapeutic index, compared with stochastic conjugation technology. This is the first report of the therapeutic index estimation of site-specific ADCs produced by utilizing Fc affinity reagent conjugation. The described site-specific conjugation technology is a powerful platform to enable next-generation ADCs through reduced heterogeneity and enhanced therapeutic index.