Skip to main content
SpringerLink
Log in

Non-linear IV pharmacokinetics of the ATR inhibitor berzosertib (M6620) in mice

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Background

The Ataxia Telangiectasia and Rad3-related (ATR) protein complex is an apical initiator of DNA damage response pathways. Several ATR inhibitors (ATRi) are in clinical development including berzosertib (formerly M6620, VX-970). Although clinical studies have examined plasma pharmacokinetics (PK) in humans, little is known regarding dose/exposure relationships and tissue distribution. To understand these concepts, we extensively characterized the PK of berzosertib in mouse plasma and tissues.

Methods

A highly sensitive LC–MS/MS method was utilized to quantitate berzosertib in plasma and tissues. Dose proportionality was assessed in female BALB/c mice following single IV doses (2, 6, 20 or 60 mg/kg). A more extensive PK study was conducted in tumor-bearing mice following a single IV dose of 20 mg/kg to evaluate distribution to tissues. PK parameters were calculated by non-compartmental analysis (NCA). A compartmental model was developed to describe the PK behavior of berzosertib. Plasma protein binding was determined in vitro.

Results

Increased doses of berzosertib were associated with less than proportional increases in early plasma concentrations and greater than proportional increase in tissue exposure, attributable to saturation of plasma protein binding. Berzosertib extensively distributed into bone marrow, tumor, thymus, and lymph nodes, however; brain and spinal cord exposure was less than plasma.

Conclusion

The nonlinear PK of berzosertib displayed here can be attributed to saturation of plasma protein binding and occurred at concentrations close to those observed in clinical trials. Our results will help to understand preclinical pharmacodynamic and toxicity data and to inform optimal dosing and deployment of berzosertib.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Data availability

Data is provided within the manuscript or supplementary information files.

Abbreviations

ATR:

Ataxia telangiectasia and Rad3-related

ATRi:

Ataxia telangiectasia and Rad3-related inhibitor

DDR:

DNA damage response

LC–MS/MS:

Liquid chromatography-tandem mass spectrometry

LLOQ:

Lower limit of quantitation

PK:

Pharmacokinetics

NCA:

Noncompartmental Analysis

Cmax :

Maximum concentration

Tmax :

Time to maximum concentration

Tlast :

Time of last measurable concentration

CL:

Clearance

Vd :

Volume of distribution

T1/2 :

Half-life

AUC:

Area under the curve

Fu,p :

Fraction unbound in plasma

References

  1. Jackson SP, Bartek J (2009) The DNA-damage response in human biology and disease. Nature 461(7267):1071–1078. https://doi.org/10.1038/nature08467

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Ciccia A, Elledge SJ (2010) The DNA damage response: making it safe to play with knives. Mol Cell 40(2):179–204. https://doi.org/10.1016/j.molcel.2010.09.019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Liao H, Ji F, Helleday T, Ying S (2018) Mechanisms for stalled replication fork stabilization: new targets for synthetic lethality strategies in cancer treatments. EMBO Rep. https://doi.org/10.15252/embr.201846263

    Article  PubMed  PubMed Central  Google Scholar 

  4. Smith HL, Southgate H, Tweddle DA, Curtin NJ (2020) DNA damage checkpoint kinases in cancer. Expert Rev Mol Med. https://doi.org/10.1017/erm.2020.3

    Article  PubMed  Google Scholar 

  5. Hoffmann I, Draetta G, Karsenti E (1994) Activation of the phosphatase activity of human cdc25A by a cdk2-cyclin E dependent phosphorylation at the G1/S transition. EMBO J 13(18):4302–4310. https://doi.org/10.1002/j.1460-2075.1994.tb06750.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Bartek J, Lukas J (2003) Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell 3(5):421–429. https://doi.org/10.1016/s1535-6108(03)00110-7

    Article  CAS  PubMed  Google Scholar 

  7. Vendetti FP, Pandya P, Clump DA, Schamus-Haynes S, Tavakoli M, diMayorca M, Islam NM, Chang J, Delgoffe GM, Beumer JH, Bakkenist CJ (2023) The schedule of ATR inhibitor AZD6738 can potentiate or abolish antitumor immune responses to radiotherapy. JCI Insight. https://doi.org/10.1172/jci.insight.165615

    Article  PubMed  PubMed Central  Google Scholar 

  8. Vendetti FP, Karukonda P, Clump DA, Teo T, Lalonde R, Nugent K, Ballew M, Kiesel BF, Beumer JH, Sarkar SN, Conrads TP, O’Connor MJ, Ferris RL, Tran PT, Delgoffe GM, Bakkenist CJ (2018) ATR kinase inhibitor AZD6738 potentiates CD8+ T cell-dependent antitumor activity following radiation. J Clin Invest 128(9):3926–3940. https://doi.org/10.1172/JCI96519

    Article  PubMed  PubMed Central  Google Scholar 

  9. Sheng H, Huang Y, Xiao Y, Zhu Z, Shen M, Zhou P, Guo Z, Wang J, Wang H, Dai W, Zhang W, Sun J, Cao C (2020) ATR inhibitor AZD6738 enhances the antitumor activity of radiotherapy and immune checkpoint inhibitors by potentiating the tumor immune microenvironment in hepatocellular carcinoma. J Immunother Cancer. https://doi.org/10.1136/jitc-2019-000340

    Article  PubMed  PubMed Central  Google Scholar 

  10. Reaper PM, Griffiths MR, Long JM, Charrier JD, Maccormick S, Charlton PA, Golec JM, Pollard JR (2011) Selective killing of ATM- or p53-deficient cancer cells through inhibition of ATR. Nat Chem Biol 7(7):428–430. https://doi.org/10.1038/nchembio.573

    Article  CAS  PubMed  Google Scholar 

  11. Fokas E, Prevo R, Pollard JR, Reaper PM, Charlton PA, Cornelissen B, Vallis KA, Hammond EM, Olcina MM, Gillies McKenna W, Muschel RJ, Brunner TB (2012) Targeting ATR in vivo using the novel inhibitor VE-822 results in selective sensitization of pancreatic tumors to radiation. Cell Death Dis 3(12):e441. https://doi.org/10.1038/cddis.2012.181

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Wilson Z, Odedra R, Wallez Y, Wijnhoven PWG, Hughes AM, Gerrard J, Jones GN, Bargh-Dawson H, Brown E, Young LA, O’Connor MJ, Lau A (2022) ATR inhibitor AZD6738 (ceralasertib) exerts antitumor activity as a monotherapy and in combination with chemotherapy and the PARP inhibitor olaparib. Cancer Res 82(6):1140–1152. https://doi.org/10.1158/0008-5472.CAN-21-2997

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Kiesel BF, Guo J, Parise RA, Venkataramanan R, Clump DA, Bakkenist CJ, Beumer JH (2022) Dose-dependent bioavailability and tissue distribution of the ATR inhibitor AZD6738 (ceralasertib) in mice. Cancer Chemother Pharmacol 89(2):231–242. https://doi.org/10.1007/s00280-021-04388-x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Kiesel BF, Deppas JJ, Guo J, Parise RA, Clump DA, Bakkenist CJ, Beumer JH (2022) Dose-dependent bioavailability, absorption-rate limited elimination, and tissue distribution of the ATR inhibitor BAY-1895344 (elimusertib) in mice. Cancer Chemother Pharmacol 89(6):795–807. https://doi.org/10.1007/s00280-022-04436-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kiesel BF, Scemama J, Parise RA, Villaruz L, Iffland A, Doyle A, Ivy P, Chu E, Bakkenist CJ, Beumer JH (2017) LC-MS/MS assay for the quantitation of the ATR kinase inhibitor VX-970 in human plasma. J Pharm Biomed Anal 146:244–250. https://doi.org/10.1016/j.jpba.2017.08.037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Bailer AJ (1988) Testing for the equality of area under the curves when using destructive measurement techniques. J Pharmacokinet Biopharm 16(3):303–309. https://doi.org/10.1007/BF01062139

    Article  CAS  PubMed  Google Scholar 

  17. Hummel J, McKendrick S, Brindley C, French R (2009) Exploratory assessment of dose proportionality: review of current approaches and proposal for a practical criterion. Pharm Stat 8(1):38–49. https://doi.org/10.1002/pst.326

    Article  PubMed  Google Scholar 

  18. Zaias J, Mineau M, Cray C, Yoon D, Altman NH (2009) Reference values for serum proteins of common laboratory rodent strains. J Am Assoc Lab Anim Sci 48(4):387–390

    CAS  PubMed  PubMed Central  Google Scholar 

  19. D’Argenio DZS, A; Wang, X, (2009) ADAPT 5 user’s guide: Pharmacokinetic/Pharmacodynamic systems analysis software. Biomedical Simulations Resource, Los Angeles

    Google Scholar 

  20. U.S. Department of Health and Human Services Food and Drug Administration (2018) Guidance for Industry-Bioanalytical Method Validation. U.S.Department of Health and Human Services; Food and Drug Administration; Center for Drug Evaluation and Research (CDER); Center for Veterinary Medicine (CVM)

  21. Yap TA, O’Carrigan B, Penney MS, Lim JS, Brown JS, de Miguel Luken MJ, Tunariu N, Perez-Lopez R, Rodrigues DN, Riisnaes R, Figueiredo I, Carreira S, Hare B, McDermott K, Khalique S, Williamson CT, Natrajan R, Pettitt SJ, Lord CJ, Banerji U, Pollard J, Lopez J, de Bono JS (2020) Phase I trial of first-in-class ATR inhibitor M6620 (VX-970) as monotherapy or in combination with carboplatin in patients with advanced solid tumors. J Clin Oncol 38(27):3195–3204. https://doi.org/10.1200/JCO.19.02404

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Talele S, Zhang W, Burgenske DM, Kim M, Mohammad AS, Dragojevic S, Gupta SK, Bindra RS, Sarkaria JN, Elmquist WF (2021) Brain distribution of berzosertib: an ataxia telangiectasia and Rad3-related protein inhibitor for the treatment of glioblastoma. J Pharmacol Exp Ther 379(3):343–357. https://doi.org/10.1124/jpet.121.000845

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Pal SK, Frankel PH, Mortazavi A, Milowsky M, Vaishampayan U, Parikh M, Lyou Y, Weng P, Parikh R, Teply B, Dreicer R, Emamekhoo H, Michaelson D, Hoimes C, Zhang T, Srinivas S, Kim WY, Cui Y, Newman E, Lara PN Jr (2021) Effect of cisplatin and gemcitabine with or without berzosertib in patients with advanced urothelial carcinoma: a phase 2 randomized clinical trial. JAMA Oncol 7(10):1536–1543. https://doi.org/10.1001/jamaoncol.2021.3441

    Article  PubMed  Google Scholar 

  24. Konstantinopoulos PA, Cheng SC, Wahner Hendrickson AE, Penson RT, Schumer ST, Doyle LA, Lee EK, Kohn EC, Duska LR, Crispens MA, Olawaiye AB, Winer IS, Barroilhet LM, Fu S, McHale MT, Schilder RJ, Farkkila A, Chowdhury D, Curtis J, Quinn RS, Bowes B, D’Andrea AD, Shapiro GI, Matulonis UA (2020) Berzosertib plus gemcitabine versus gemcitabine alone in platinum-resistant high-grade serous ovarian cancer: a multicentre, open-label, randomised, phase 2 trial. Lancet Oncol 21(7):957–968. https://doi.org/10.1016/S1470-2045(20)30180-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Middleton MR, Dean E, Evans TRJ, Shapiro GI, Pollard J, Hendriks BS, Falk M, Diaz-Padilla I, Plummer R (2021) Phase 1 study of the ATR inhibitor berzosertib (formerly M6620, VX-970) combined with gemcitabine +/- cisplatin in patients with advanced solid tumours. Br J Cancer 125(4):510–519. https://doi.org/10.1038/s41416-021-01405-x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Telli ML, Tolaney SM, Shapiro GI, Middleton M, Lord SR, Arkenau HT, Tutt A, Abramson V, Dean E, Haddad TC, Wesolowski R, Ferrer-Playan J, Goddemeier T, Grombacher T, Dong J, Fleuranceau-Morel P, Diaz-Padilla I, Plummer R (2022) Phase 1b study of berzosertib and cisplatin in patients with advanced triple-negative breast cancer. NPJ Breast Cancer 8(1):45. https://doi.org/10.1038/s41523-022-00406-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Thomas A, Redon CE, Sciuto L, Padiernos E, Ji J, Lee MJ, Yuno A, Lee S, Zhang Y, Tran L, Yutzy W, Rajan A, Guha U, Chen H, Hassan R, Alewine CC, Szabo E, Bates SE, Kinders RJ, Steinberg SM, Doroshow JH, Aladjem MI, Trepel JB, Pommier Y (2018) Phase I study of ATR Inhibitor M6620 in combination with topotecan in patients with advanced solid tumors. J Clin Oncol 36(16):1594–1602. https://doi.org/10.1200/JCO.2017.76.6915

    Article  CAS  PubMed  Google Scholar 

  28. U.S. Department of Health and Human Services Food and Drug Administration (2005) Guidance for Industry-Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers. U.S. Department of Health and Human Services; Food and Drug Administration; Center for Drug Evaluation and Research (CDER)

  29. Plummer R, Dean E, Arkenau HT, Redfern C, Spira AI, Melear JM, Chung KY, Ferrer-Playan J, Goddemeier T, Locatelli G, Dong J, Fleuranceau-Morel P, Diaz-Padilla I, Shapiro GI (2022) A phase 1b study evaluating the safety and preliminary efficacy of berzosertib in combination with gemcitabine in patients with advanced non-small cell lung cancer. Lung Cancer 163:19–26. https://doi.org/10.1016/j.lungcan.2021.11.011

    Article  CAS  PubMed  Google Scholar 

  30. Takahashi N, Hao Z, Villaruz LC, Zhang J, Ruiz J, Petty WJ, Mamdani H, Riess JW, Nieva J, Pachecho JM, Fuld AD, Shum E, Chauhan A, Nichols S, Shimellis H, McGlone J, Sciuto L, Pinkiert D, Graham C, Shelat M, Kattappuram R, Abel M, Schroeder B, Upadhyay D, Krishnamurthy M, Sharma AK, Kumar R, Malin J, Schultz CW, Goyal S, Redon CE, Pommier Y, Aladjem MI, Gore SD, Steinberg SM, Vilimas R, Desai P, Thomas A (2023) Berzosertib plus topotecan vs topotecan alone in patients with relapsed small cell lung cancer: a randomized clinical trial. JAMA Oncol 9(12):1669–1677. https://doi.org/10.1001/jamaoncol.2023.4025

    Article  PubMed  Google Scholar 

  31. Shapiro GI, Wesolowski R, Devoe C, Lord S, Pollard J, Hendriks BS, Falk M, Diaz-Padilla I, Plummer R, Yap TA (2021) Phase 1 study of the ATR inhibitor berzosertib in combination with cisplatin in patients with advanced solid tumours. Br J Cancer 125(4):520–527. https://doi.org/10.1038/s41416-021-01406-w

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported in-part by the National Institutes of Health grants R50CA211241 and R01CA266172. This project was funded, in-part, under a CURE Grant with the Pennsylvania Department of Health. The Department specifically disclaims responsibility for any analyses, interpretations, or conclusions. Research reported in this publication was also supported in-part by the National Center For Advancing Translational Sciences of the National Institutes of Health under Award Number TL1TR001858. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. JJD was also supported in-part by a scholarship from the Community Foundation of Warren County (PA). This project used the UPMC Hillman Cancer Center, Cancer Pharmacokinetics and Pharmacodynamics Facility (CPPF), Animal Facility and used Hillman shared resources supported in part by award P30CA047904. Phoenix WinNonlin was generously provided to the University of Pittsburgh School of Pharmacy by Certara, Inc., through the Center of Excellence Program for academic institutions.

Funding

National Institutes of Health, TL1TR001858, TL1TR001858,R50CA211241, R01CA266172, P30CA047904

Author information

Authors and Affiliations

  1. Cancer Therapeutics Program, UPMC Hillman Cancer Center, University of Pittsburgh, Room G27e 5117 Centre Ave, Pittsburgh, PA, 15213, USA

    Joshua J. Deppas, Brian F. Kiesel, Jianxia Guo, Robert A. Parise & Jan H. Beumer

  2. Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, PA, USA

    Joshua J. Deppas, Brian F. Kiesel & Jan H. Beumer

  3. Department of Radiation Oncology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    D. Andy Clump & Christopher J. Bakkenist

  4. Department of Biomedical Engineering, University of Southern California, Los Angeles, CA, USA

    David Z. D’Argenio

  5. Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Christopher J. Bakkenist

  6. Division of Hematology/Oncology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA

    Jan H. Beumer

Authors
  1. Joshua J. Deppas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Brian F. Kiesel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianxia Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Robert A. Parise
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. D. Andy Clump
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. David Z. D’Argenio
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christopher J. Bakkenist
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jan H. Beumer
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Joshua J. Deppas: Conceptualization, Investigation, Writing - Original Draft, Writing - Review & Editing, Project administration Brian F. Kiesel: Conceptualization, Investigation, Writing - Original Draft, Writing - Review & Editing, Project administration Jianxia Guo: Investigation, Writing - Review & Editing, Project administration Robert. A Parise: Investigation, Writing - Review & Editing D. Andy Clump: Investigation, Writing - Review & Editing David Z. D’Argenio: Conceptualization, Investigation, Writing - Review & Editing Christopher J. Bakkenist: Conceptualization, Writing - Review & Editing, Project administration Jan H. Beumer: Conceptualization, Investigation, Writing - Original Draft, Writing - Review & Editing, Project administration

Corresponding author

Correspondence to Jan H. Beumer.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 455 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Deppas, J.J., Kiesel, B.F., Guo, J. et al. Non-linear IV pharmacokinetics of the ATR inhibitor berzosertib (M6620) in mice. Cancer Chemother Pharmacol (2024). https://doi.org/10.1007/s00280-024-04675-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00280-024-04675-3

Keywords

Search

Navigation