Development of novel ADCs: conjugation of tubulysin analogs to trastuzumab monitored by dual radiolabeling

Ruth Cohen, Danielle J. Vugts, Gerard W.M. Visser, Marijke Stigter-van Walsum, Marije Bolijn, Marco Spiga, Paolo Lazzari, Sreejith P. Shankar, Monica Sani, Matteo Zanda, Guus A.M.S. Van Dongen

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Abstract

Tubulysins are highly toxic tubulin-targeting agents with a narrow therapeutic window that are interesting for application in antibody–drug conjugates (ADC). For full control over drug–antibody ratio (DAR) and the effect thereof on pharmacokinetics and tumor targeting, a dual-labeling approach was developed, wherein the drug, tubulysin variants, and the antibody, the anti-HER2 monoclonal antibody (mAb) trastuzumab, are radiolabeled. 131I-radioiodination of two synthetic tubulysin A analogues, the less potent TUB-OH (IC50 > 100 nmol/L) and the potent TUB-OMOM (IC50, ∼1 nmol/L), and their direct covalent conjugation to 89Zr-trastuzumab were established. Radioiodination of tubulysins was 92% to 98% efficient and conversion to N-hydroxysuccinimide (NHS) esters more than 99%; esters were isolated in an overall yield of 68% ± 5% with radiochemical purity of more than 99.5%. Conjugation of 131I-tubulysin–NHS esters to 89Zr-trastuzumab was 45% to 55% efficient, resulting in ADCs with 96% to 98% radiochemical purity after size-exclusion chromatography. ADCs were evaluated for their tumor-targeting potential and antitumor effects in nude mice with tumors that were sensitive or resistant to trastuzumab, using ado-trastuzumab emtansine as a reference. ADCs appeared stable in vivo. An average DAR of 2 and 4 conferred pharmacokinetics and tumor-targeting behavior similar to parental trastuzumab. Efficacy studies using single-dose TUB-OMOM–trastuzumab (DAR 4) showed dose-dependent antitumor effects, including complete tumor eradications in trastuzumab-sensitive tumors in vivo. TUB-OMOM–trastuzumab (60 mg/kg) displayed efficacy similar to ado-trastuzumab emtansine (15 mg/kg) yet more effective than trastuzumab. Our findings illustrate the potential of synthetic tubulysins in ADCs for cancer treatment. Cancer Res; 74(20); 5700–10. ©2014 AACR.
Original languageEnglish
Pages (from-to)5700-5710
JournalCancer Research
Volume74
Issue number20
Early online date21 Aug 2014
DOIs
Publication statusPublished - 15 Oct 2014

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Neoplasms
Esters
Inhibitory Concentration 50
Pharmacokinetics
Trastuzumab
Poisons
Tubulin
Nude Mice
Gel Chromatography
Anti-Idiotypic Antibodies
Monoclonal Antibodies
Pharmaceutical Preparations
ado-trastuzumab emtansine
zirconium-89-trastuzumab
Therapeutics

Cite this

Cohen, R., Vugts, D. J., Visser, G. W. M., Stigter-van Walsum, M., Bolijn, M., Spiga, M., ... Van Dongen, G. A. M. S. (2014). Development of novel ADCs: conjugation of tubulysin analogs to trastuzumab monitored by dual radiolabeling. Cancer Research, 74(20), 5700-5710. https://doi.org/10.1158/0008-5472.CAN-14-1141

Development of novel ADCs : conjugation of tubulysin analogs to trastuzumab monitored by dual radiolabeling. / Cohen, Ruth; Vugts, Danielle J.; Visser, Gerard W.M.; Stigter-van Walsum, Marijke; Bolijn, Marije; Spiga, Marco; Lazzari, Paolo; Shankar, Sreejith P.; Sani, Monica; Zanda, Matteo; Van Dongen, Guus A.M.S.

In: Cancer Research, Vol. 74, No. 20, 15.10.2014, p. 5700-5710.

Research output: Contribution to journalArticle

Cohen, R, Vugts, DJ, Visser, GWM, Stigter-van Walsum, M, Bolijn, M, Spiga, M, Lazzari, P, Shankar, SP, Sani, M, Zanda, M & Van Dongen, GAMS 2014, 'Development of novel ADCs: conjugation of tubulysin analogs to trastuzumab monitored by dual radiolabeling', Cancer Research, vol. 74, no. 20, pp. 5700-5710. https://doi.org/10.1158/0008-5472.CAN-14-1141
Cohen R, Vugts DJ, Visser GWM, Stigter-van Walsum M, Bolijn M, Spiga M et al. Development of novel ADCs: conjugation of tubulysin analogs to trastuzumab monitored by dual radiolabeling. Cancer Research. 2014 Oct 15;74(20):5700-5710. https://doi.org/10.1158/0008-5472.CAN-14-1141
Cohen, Ruth ; Vugts, Danielle J. ; Visser, Gerard W.M. ; Stigter-van Walsum, Marijke ; Bolijn, Marije ; Spiga, Marco ; Lazzari, Paolo ; Shankar, Sreejith P. ; Sani, Monica ; Zanda, Matteo ; Van Dongen, Guus A.M.S. / Development of novel ADCs : conjugation of tubulysin analogs to trastuzumab monitored by dual radiolabeling. In: Cancer Research. 2014 ; Vol. 74, No. 20. pp. 5700-5710.
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