ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Figure 1Loading Img

Preclinical Development of an anti-5T4 Antibody–Drug Conjugate: Pharmacokinetics in Mice, Rats, and NHP and Tumor/Tissue Distribution in Mice

View Author Information
Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Pearl River, New York 10965, United States
Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Groton, Connecticut 06340, United States
§ Pharmacokinetics, Dynamics and Metabolism, Pfizer Inc., Andover, Massachusetts 01810, United States
Oncology Research Unit, Pfizer Inc., Pearl River, New York 10965, United States
*E-mail: [email protected]. Tel: 1 845 602 2667.
Cite this: Bioconjugate Chem. 2015, 26, 11, 2223–2232
Publication Date (Web):July 16, 2015
Copyright © 2015 American Chemical Society

    Article Views





    Other access options
    Supporting Info (1)»


    Abstract Image

    The pharmacokinetics of an antibody (huA1)–drug (auristatin microtubule disrupting MMAF) conjugate, targeting 5T4-expressing cells, were characterized during the discovery and development phases in female nu/nu mice and cynomolgus monkeys after a single dose and in S-D rats and cynomolgus monkeys from multidose toxicity studies. Plasma/serum samples were analyzed using an ELISA-based method for antibody and conjugate (ADC) as well as for the released payload using an LC-MS/MS method. In addition, the distribution of the Ab, ADC, and released payload (cys-mcMMAF) was determined in a number of tissues (tumor, lung, liver, kidney, and heart) in two tumor mouse models (H1975 and MDA-MB-361-DYT2 models) using similar LBA and LC-MS/MS methods. Tissue distribution studies revealed preferential tumor distribution of cys-mcMMAF and its relative specificity to the 5T4 target containing tissue (tumor). Single dose studies suggests lower CL values at the higher doses in mice, although a linear relationship was seen in cynomolgus monkeys at doses from 0.3 to 10 mg/kg with no evidence of TMDD. Evaluation of DAR (drug–antibody ratio) in cynomolgus monkeys (at 3 mg/kg) indicated that at least half of the payload was still on the ADC 1 to 2 weeks after IV dosing. After multiple doses, the huA1 and conjugate data in rats and monkeys indicate that exposure (AUC) increases with increasing dose in a linear fashion. Systemic exposure (as assessed by Cmax and AUC) of the released payload increased with increasing dose, although exposure was very low and its pharmacokinetics appeared to be formation rate limited. The incidence of ADA was generally low in rats and monkeys. We will discuss cross species comparison, relationships between the Ab, ADC, and released payload exposure after multiple dosing, and insights into the distribution of this ADC with a focus on experimental design as a way to address or bypass apparent obstacles and its integration into predictive models.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Supporting Information

    Jump To

    Supplemental figures. The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.bioconjchem.5b00205.

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system:

    Cited By

    This article is cited by 20 publications.

    1. Ettore Gilardoni, Aureliano Zana, Andrea Galbiati, Theo Sturm, Jacopo Millul, Samuele Cazzamalli, Dario Neri, Riccardo Stucchi. Mass Spectrometry-Based Method for the Determination of the Biodistribution of Tumor-Targeting Small Molecule–Metal Conjugates. Analytical Chemistry 2022, 94 (30) , 10715-10721.
    2. Yan Zheng, Ruolin Xu, Hong Cheng, Wanyi Tai. Mono-amino acid linkers enable highly potent small molecule-drug conjugates by conditional release. Molecular Therapy 2024, 22
    3. Siddhanth Hejmady, Rajesh Pradhan, Shobha Kumari, Meghna Pandey, Sunil K Dubey, Rajeev Taliyan. Pharmacokinetics and toxicity considerations for antibody–drug conjugates: an overview. Bioanalysis 2023, 15 (19) , 1193-1202.
    4. Aureliano Zana, Andrea Galbiati, Ettore Gilardoni, Matilde Bocci, Jacopo Millul, Theo Sturm, Riccardo Stucchi, Abdullah Elsayed, Lisa Nadal, Martina Cirillo, Wolfgang Roll, Lars Stegger, Inga Asmus, Philipp Backhaus, Michael Schäfers, Dario Neri, Samuele Cazzamalli. Fibroblast Activation Protein Triggers Release of Drug Payload from Non-internalizing Small Molecule Drug Conjugates in Solid Tumors. Clinical Cancer Research 2022, 28 (24) , 5440-5454.
    5. Beverly A. Teicher, Joel Morris. Antibody-drug Conjugate Targets, Drugs, and Linkers. Current Cancer Drug Targets 2022, 22 (6) , 463-529.
    6. Iftekhar Mahmood. A Simple Method for the Prediction of Human Concentration–Time Profiles and Pharmacokinetics of Antibody–Drug Conjugates (ADC) from Rats or Monkeys. Antibodies 2022, 11 (2) , 42.
    7. Breanna S. Vollmar, Chris Frantz, Melissa M. Schutten, Fiona Zhong, Geoffrey del Rosario, Mary Ann T. Go, Shang-Fan Yu, Douglas D. Leipold, Amrita V. Kamath, Carl Ng, Keyang Xu, Josefa dela Cruz-Chuh, Katherine R. Kozak, Jinhua Chen, Zijin Xu, John Wai, Pragya Adhikari, Hans K. Erickson, Peter S. Dragovich, Andrew G. Polson, Thomas H. Pillow. Calicheamicin Antibody–Drug Conjugates with Improved Properties. Molecular Cancer Therapeutics 2021, 20 (6) , 1112-1120.
    8. Iftekhar Mahmood. Interspecies Scaling of Antibody–Drug Conjugates (ADC) for the Prediction of Human Clearance. Antibodies 2021, 10 (1) , 1.
    9. Héloïse Cahuzac, Laurent Devel. Analytical Methods for the Detection and Quantification of ADCs in Biological Matrices. Pharmaceuticals 2020, 13 (12) , 462.
    10. Changyong Yang, Xiaoping Zhao, Xing Sun, Jinlong Li, Weiqiang Wang, Lianshan Zhang, Shaohua Gou. Preclinical pharmacokinetics of a novel anti-c-Met antibody–drug conjugate, SHR-A1403, in rodents and non-human primates. Xenobiotica 2019, 49 (9) , 1097-1105.
    11. Giuseppe Merlino, Alessio Fiascarelli, Mario Bigioni, Alessandro Bressan, Corrado Carrisi, Daniela Bellarosa, Massimiliano Salerno, Rossana Bugianesi, Rosanna Manno, Cristina Bernadó Morales, Joaquin Arribas, Arnima Bisht, Angelo Kaplan, Robert Boyd, Uyen T. Do, Sudha Swaminathan, Nickolas Attanasio, San Lin Lou, Rachel L. Dusek, James E. Ackroyd, Phuoc Huy Pham, Rahel Awdew, Dee Aud, Michael Trang, Carmel M. Lynch, Jonathan Terrett, Keith E. Wilson, Christian Rohlff, Stefano Manzini, Andrea Pellacani, Monica Binaschi. MEN1309/OBT076, a First-In-Class Antibody–Drug Conjugate Targeting CD205 in Solid Tumors. Molecular Cancer Therapeutics 2019, 18 (9) , 1533-1543.
    12. Y. Louise Wan, Puja Sapra, James Bolton, Jia Xin Chua, Lindy G. Durrant, Peter L. Stern. Combination Treatment with an Antibody–Drug Conjugate (A1mcMMAF) Targeting the Oncofetal Glycoprotein 5T4 and Carboplatin Improves Survival in a Xenograft Model of Ovarian Cancer. Targeted Oncology 2019, 14 (4) , 465-477.
    13. Chao Li, Linlin Dong, Afrand Kamali, Hiroshi Sugimoto, Kojo Abdul-Hadi, Susan Chen, Adnan Abu-Yousif, Mark G. Qian. An LC/MS based method to quantify DNA adduct in tumor and organ tissues. Analytical Biochemistry 2019, 568 , 1-6.
    14. Brandon Bumbaca, Zhe Li, Dhaval K. Shah. Pharmacokinetics of protein and peptide conjugates. Drug Metabolism and Pharmacokinetics 2019, 34 (1) , 42-54.
    15. David A. Eavarone, Linah Al-Alem, Alexey Lugovskoy, Jillian M. Prendergast, Rawan I. Nazer, Jenna N. Stein, Daniel T. Dransfield, Jeff Behrens, Bo R. Rueda, . Humanized anti-Sialyl-Tn antibodies for the treatment of ovarian carcinoma. PLOS ONE 2018, 13 (7) , e0201314.
    16. Cong Wei, Dian Su, Jian Wang, Wenying Jian, Donglu Zhang. LC–MS Challenges in Characterizing and Quantifying Monoclonal Antibodies (mAb) and Antibody-Drug Conjugates (ADC) in Biological Samples. Current Pharmacology Reports 2018, 4 (1) , 45-63.
    17. Mary Jane Masson Hinrichs, Pauline M. Ryan, Bo Zheng, Shameen Afif-Rider, Xiang Qing Yu, Michele Gunsior, Haihong Zhong, Jay Harper, Binyam Bezabeh, Kapil Vashisht, Marlon Rebelatto, Molly Reed, Patricia C. Ryan, Shannon Breen, Neki Patel, Cui Chen, Luke Masterson, Arnaud Tiberghien, Phillip W. Howard, Nazzareno Dimasi, Rakesh Dixit. Fractionated Dosing Improves Preclinical Therapeutic Index of Pyrrolobenzodiazepine-Containing Antibody Drug Conjugates. Clinical Cancer Research 2017, 23 (19) , 5858-5868.
    18. Xiaogang Han, Steven Hansel, Lindsay King. Bioanalytical Strategies Enabling Successful ADC Translation. 2016, 177-205.
    19. Anand Giddabasappa, Vijay R. Gupta, Rand Norberg, Parul Gupta, Mary E. Spilker, Joann Wentland, Brian Rago, Jeetendra Eswaraka, Mauricio Leal, Puja Sapra. Biodistribution and Targeting of Anti-5T4 Antibody–Drug Conjugate Using Fluorescence Molecular Tomography. Molecular Cancer Therapeutics 2016, 15 (10) , 2530-2540.
    20. D. Zhang, S.-F. Yu, Y. Ma, K. Xu, P. S. Dragovich, T. H. Pillow, L. Liu, G. Del Rosario, J. He, Z. Pei, J. D. Sadowsky, H. K. Erickson, C. E. C. A. Hop, S. C. Khojasteh. Chemical Structure and Concentration of Intratumor Catabolites Determine Efficacy of Antibody Drug Conjugates. Drug Metabolism and Disposition 2016, 44 (9) , 1517-1523.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect