In vivo characterization of platinum (II)-based linker technology for the development of antibody-drug conjugates: Taking advantage of dual labeling with 195mPt and 89Zr

Joey A. Muns, Veronica Montserrat, Hendrik Jan Houthoff, Karlijn Codée-Van Der Schilden, Oene Zwaagstra, Niels J. Sijbrandi, Eugen Merkul, Guus A.M.S. Van Dongen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Linker instability and impaired tumor targeting can affect the tolerability and efficacy of antibody-drug conjugates (ADCs). To improve these ADC characteristics, we recently described the use of a metal-organic linker, [ethylenediamineplatinum(II)]21, herein called Lx. Initial therapy studies in xenograft-bearing mice revealed that trastuzumab-Lx-auristatin F (AF) outperformed its maleimide benchmark trastuzumab-mal-AF and the Food and Drug Administration- approved ado-trastuzumab emtansine, both containing conventional linkers. In this study, we aimed to characterize Lx-based ADCs for in vivo stability and tumor targeting using 195mPt and 89Zr. Methods: The γ-emitter 195mPt was used to produce the radiolabeled Lx [195mPt]Lx. 89Zr-Desferrioxamine (89Zr-DFO) was conjugated to trastuzumab either via [195mPt]Lx (to histidine residues) or conventionally (to lysine residues) in order to monitor the biodistribution of antibody, payload, and linker separately. Linker stability was determined by evaluating the following ADCs for biodistribution in NCI-N87 xenograft-bearing nude mice 72 h after injection: Trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx- AF, and 89Zr-DFO-(Lys)trastuzumab (control), all having drug-toantibody ratios (DARs) of 2.2-2.5. To assess the influence of DAR on biodistribution, 89Zr-DFO-(Lys)trastuzumab-Lx-AF with an AF-to-antibody ratio of 0, 2.6, or 5.2 was evaluated 96 h after injection. Results: Similar biodistributions were observed for trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx-AF, and 89Zr-DFO-(Lys)trastuzumab irrespective of the isotope used for biodistribution assessment. The fact that Lx follows the antibody biodistribution indicates that the payload-Lx bond is stable in vivo. Uptake of the 3 conjugates, as percentage injected dose (%ID) per gram of tissue, was about 30 %ID/g in tumor tissue but less than 10 % ID/g in most healthy tissues. Trastuzumab-[195mPt]Lx-AF (DAR 2.2) showed a tendency toward faster blood clearance and an elevated liver uptake, which increased significantly to 28.1 ± 4.2 %ID/g at a higher DAR of 5.2, as revealed from the biodistribution and PET imaging studies. Conclusion: As shown by 195mPt/89Zr labeling, ADCs containing the Lx linker are stable in vivo. In the case of trastuzumab-Lx-AF (DARs 2.2 and 2.6), an unimpaired biodistribution was demonstrated.

Original languageEnglish
Pages (from-to)1146-1151
Number of pages6
JournalJournal of Nuclear Medicine
Volume59
Issue number7
DOIs
Publication statusPublished - 1 Jul 2018

Cite this

@article{b9ab39cb14e74615be59042bb4e34442,
title = "In vivo characterization of platinum (II)-based linker technology for the development of antibody-drug conjugates: Taking advantage of dual labeling with 195mPt and 89Zr",
abstract = "Linker instability and impaired tumor targeting can affect the tolerability and efficacy of antibody-drug conjugates (ADCs). To improve these ADC characteristics, we recently described the use of a metal-organic linker, [ethylenediamineplatinum(II)]21, herein called Lx. Initial therapy studies in xenograft-bearing mice revealed that trastuzumab-Lx-auristatin F (AF) outperformed its maleimide benchmark trastuzumab-mal-AF and the Food and Drug Administration- approved ado-trastuzumab emtansine, both containing conventional linkers. In this study, we aimed to characterize Lx-based ADCs for in vivo stability and tumor targeting using 195mPt and 89Zr. Methods: The γ-emitter 195mPt was used to produce the radiolabeled Lx [195mPt]Lx. 89Zr-Desferrioxamine (89Zr-DFO) was conjugated to trastuzumab either via [195mPt]Lx (to histidine residues) or conventionally (to lysine residues) in order to monitor the biodistribution of antibody, payload, and linker separately. Linker stability was determined by evaluating the following ADCs for biodistribution in NCI-N87 xenograft-bearing nude mice 72 h after injection: Trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx- AF, and 89Zr-DFO-(Lys)trastuzumab (control), all having drug-toantibody ratios (DARs) of 2.2-2.5. To assess the influence of DAR on biodistribution, 89Zr-DFO-(Lys)trastuzumab-Lx-AF with an AF-to-antibody ratio of 0, 2.6, or 5.2 was evaluated 96 h after injection. Results: Similar biodistributions were observed for trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx-AF, and 89Zr-DFO-(Lys)trastuzumab irrespective of the isotope used for biodistribution assessment. The fact that Lx follows the antibody biodistribution indicates that the payload-Lx bond is stable in vivo. Uptake of the 3 conjugates, as percentage injected dose ({\%}ID) per gram of tissue, was about 30 {\%}ID/g in tumor tissue but less than 10 {\%} ID/g in most healthy tissues. Trastuzumab-[195mPt]Lx-AF (DAR 2.2) showed a tendency toward faster blood clearance and an elevated liver uptake, which increased significantly to 28.1 ± 4.2 {\%}ID/g at a higher DAR of 5.2, as revealed from the biodistribution and PET imaging studies. Conclusion: As shown by 195mPt/89Zr labeling, ADCs containing the Lx linker are stable in vivo. In the case of trastuzumab-Lx-AF (DARs 2.2 and 2.6), an unimpaired biodistribution was demonstrated.",
keywords = "Pt, Zr, ADC radiolabeling, Antibody-drug conjugates, Lx linker",
author = "Muns, {Joey A.} and Veronica Montserrat and Houthoff, {Hendrik Jan} and {Cod{\'e}e-Van Der Schilden}, Karlijn and Oene Zwaagstra and Sijbrandi, {Niels J.} and Eugen Merkul and {Van Dongen}, {Guus A.M.S.}",
year = "2018",
month = "7",
day = "1",
doi = "10.2967/jnumed.117.206672",
language = "English",
volume = "59",
pages = "1146--1151",
journal = "Journal of Nuclear Medicine",
issn = "0161-5505",
publisher = "Society of Nuclear Medicine Inc.",
number = "7",

}

In vivo characterization of platinum (II)-based linker technology for the development of antibody-drug conjugates : Taking advantage of dual labeling with 195mPt and 89Zr. / Muns, Joey A.; Montserrat, Veronica; Houthoff, Hendrik Jan; Codée-Van Der Schilden, Karlijn; Zwaagstra, Oene; Sijbrandi, Niels J.; Merkul, Eugen; Van Dongen, Guus A.M.S.

In: Journal of Nuclear Medicine, Vol. 59, No. 7, 01.07.2018, p. 1146-1151.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - In vivo characterization of platinum (II)-based linker technology for the development of antibody-drug conjugates

T2 - Taking advantage of dual labeling with 195mPt and 89Zr

AU - Muns, Joey A.

AU - Montserrat, Veronica

AU - Houthoff, Hendrik Jan

AU - Codée-Van Der Schilden, Karlijn

AU - Zwaagstra, Oene

AU - Sijbrandi, Niels J.

AU - Merkul, Eugen

AU - Van Dongen, Guus A.M.S.

PY - 2018/7/1

Y1 - 2018/7/1

N2 - Linker instability and impaired tumor targeting can affect the tolerability and efficacy of antibody-drug conjugates (ADCs). To improve these ADC characteristics, we recently described the use of a metal-organic linker, [ethylenediamineplatinum(II)]21, herein called Lx. Initial therapy studies in xenograft-bearing mice revealed that trastuzumab-Lx-auristatin F (AF) outperformed its maleimide benchmark trastuzumab-mal-AF and the Food and Drug Administration- approved ado-trastuzumab emtansine, both containing conventional linkers. In this study, we aimed to characterize Lx-based ADCs for in vivo stability and tumor targeting using 195mPt and 89Zr. Methods: The γ-emitter 195mPt was used to produce the radiolabeled Lx [195mPt]Lx. 89Zr-Desferrioxamine (89Zr-DFO) was conjugated to trastuzumab either via [195mPt]Lx (to histidine residues) or conventionally (to lysine residues) in order to monitor the biodistribution of antibody, payload, and linker separately. Linker stability was determined by evaluating the following ADCs for biodistribution in NCI-N87 xenograft-bearing nude mice 72 h after injection: Trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx- AF, and 89Zr-DFO-(Lys)trastuzumab (control), all having drug-toantibody ratios (DARs) of 2.2-2.5. To assess the influence of DAR on biodistribution, 89Zr-DFO-(Lys)trastuzumab-Lx-AF with an AF-to-antibody ratio of 0, 2.6, or 5.2 was evaluated 96 h after injection. Results: Similar biodistributions were observed for trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx-AF, and 89Zr-DFO-(Lys)trastuzumab irrespective of the isotope used for biodistribution assessment. The fact that Lx follows the antibody biodistribution indicates that the payload-Lx bond is stable in vivo. Uptake of the 3 conjugates, as percentage injected dose (%ID) per gram of tissue, was about 30 %ID/g in tumor tissue but less than 10 % ID/g in most healthy tissues. Trastuzumab-[195mPt]Lx-AF (DAR 2.2) showed a tendency toward faster blood clearance and an elevated liver uptake, which increased significantly to 28.1 ± 4.2 %ID/g at a higher DAR of 5.2, as revealed from the biodistribution and PET imaging studies. Conclusion: As shown by 195mPt/89Zr labeling, ADCs containing the Lx linker are stable in vivo. In the case of trastuzumab-Lx-AF (DARs 2.2 and 2.6), an unimpaired biodistribution was demonstrated.

AB - Linker instability and impaired tumor targeting can affect the tolerability and efficacy of antibody-drug conjugates (ADCs). To improve these ADC characteristics, we recently described the use of a metal-organic linker, [ethylenediamineplatinum(II)]21, herein called Lx. Initial therapy studies in xenograft-bearing mice revealed that trastuzumab-Lx-auristatin F (AF) outperformed its maleimide benchmark trastuzumab-mal-AF and the Food and Drug Administration- approved ado-trastuzumab emtansine, both containing conventional linkers. In this study, we aimed to characterize Lx-based ADCs for in vivo stability and tumor targeting using 195mPt and 89Zr. Methods: The γ-emitter 195mPt was used to produce the radiolabeled Lx [195mPt]Lx. 89Zr-Desferrioxamine (89Zr-DFO) was conjugated to trastuzumab either via [195mPt]Lx (to histidine residues) or conventionally (to lysine residues) in order to monitor the biodistribution of antibody, payload, and linker separately. Linker stability was determined by evaluating the following ADCs for biodistribution in NCI-N87 xenograft-bearing nude mice 72 h after injection: Trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx- AF, and 89Zr-DFO-(Lys)trastuzumab (control), all having drug-toantibody ratios (DARs) of 2.2-2.5. To assess the influence of DAR on biodistribution, 89Zr-DFO-(Lys)trastuzumab-Lx-AF with an AF-to-antibody ratio of 0, 2.6, or 5.2 was evaluated 96 h after injection. Results: Similar biodistributions were observed for trastuzumab-[195mPt]Lx-DFO-89Zr, trastuzumab-[195mPt]Lx-AF, and 89Zr-DFO-(Lys)trastuzumab irrespective of the isotope used for biodistribution assessment. The fact that Lx follows the antibody biodistribution indicates that the payload-Lx bond is stable in vivo. Uptake of the 3 conjugates, as percentage injected dose (%ID) per gram of tissue, was about 30 %ID/g in tumor tissue but less than 10 % ID/g in most healthy tissues. Trastuzumab-[195mPt]Lx-AF (DAR 2.2) showed a tendency toward faster blood clearance and an elevated liver uptake, which increased significantly to 28.1 ± 4.2 %ID/g at a higher DAR of 5.2, as revealed from the biodistribution and PET imaging studies. Conclusion: As shown by 195mPt/89Zr labeling, ADCs containing the Lx linker are stable in vivo. In the case of trastuzumab-Lx-AF (DARs 2.2 and 2.6), an unimpaired biodistribution was demonstrated.

KW - Pt

KW - Zr

KW - ADC radiolabeling

KW - Antibody-drug conjugates

KW - Lx linker

UR - http://www.scopus.com/inward/record.url?scp=85049402989&partnerID=8YFLogxK

U2 - 10.2967/jnumed.117.206672

DO - 10.2967/jnumed.117.206672

M3 - Article

VL - 59

SP - 1146

EP - 1151

JO - Journal of Nuclear Medicine

JF - Journal of Nuclear Medicine

SN - 0161-5505

IS - 7

ER -