Top

Published in:

Open Access 01-05-2016

Current Status: Site-Specific Antibody Drug Conjugates

Authors: Dominik Schumacher, Christian P. R. Hackenberger, Heinrich Leonhardt, Jonas Helma

Published in: Journal of Clinical Immunology | Special Issue 1/2016

Abstract

Antibody drug conjugates (ADCs), a promising class of cancer biopharmaceuticals, combine the specificity of therapeutic antibodies with the pharmacological potency of chemical, cytotoxic drugs. Ever since the first ADCs on the market, a plethora of novel ADC technologies has emerged, covering as diverse aspects as antibody engineering, chemical linker optimization and novel conjugation strategies, together aiming at constantly widening the therapeutic window for ADCs. This review primarily focuses on novel chemical and biotechnological strategies for the site-directed attachment of drugs that are currently validated for 2nd generation ADCs to promote conjugate homogeneity and overall stability.

Agarwal P, Bertozzi CR. Site-specific antibody-drug conjugates: the nexus of bioorthogonal chemistry, protein engineering, and drug development. Bioconjug Chem. 2015. doi:10.1021/bc5004982.PubMedPubMedCentral

Chari RV, Miller ML, Widdison WC. Antibody-drug conjugates: an emerging concept in cancer therapy. Angew Chem Int Ed Engl. 2014;53:3796–827. doi:10.1002/anie.201307628.CrossRefPubMed

Sochaj AM, Swiderska KW, Otlewski J. Current methods for the synthesis of homogeneous antibody-drug conjugates. Biotechnol Adv. 2015;33:775–84. doi:10.1016/j.biotechadv.2015.05.001.CrossRefPubMed

Polakis P Antibody drug conjugates for cancer therapy. Pharmacol Rev. 2016;68:3–19. doi:10.1124/pr.114.009373.CrossRefPubMed

Mathur R, Weiner GJ. Picking the optimal target for antibody-drug conjugates. Am Soc Clin Oncol Educ Book. 2013. doi:10.1200/EdBook_AM.2013.33.e103.PubMed

Slamon DJ, Leyland-Jones B, Shak S, Fuchs H, Paton V, Bajamonde A, et al. Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med. 2001;344:783–92. doi:10.1056/NEJM200103153441101.CrossRefPubMed

Hamblett KJ, Senter PD, Chace DF, Sun MM, Lenox J, Cerveny CG, et al. Effects of drug loading on the antitumor activity of a monoclonal antibody drug conjugate. Clin Cancer Res. 2004;10:7063–70. doi:10.1158/1078-0432.CCR-04-0789.CrossRefPubMed

Lyon RP, Bovee TD, Doronina SO, Burke PJ, Hunter JH, Neff-LaFord HD, et al. Reducing hydrophobicity of homogeneous antibody-drug conjugates improves pharmacokinetics and therapeutic index. Nat Biotechnol. 2015;33:733–5. doi:10.1038/nbt.3212.CrossRefPubMed

Adem YT, Schwarz KA, Duenas E, Patapoff TW, Galush WJ, Esue O. Auristatin antibody drug conjugate physical instability and the role of drug payload. Bioconjug Chem. 2014;25:656–64. doi:10.1021/bc400439x.CrossRefPubMed

10.

Zhao RY, Wilhelm SD, Audette C, Jones G, Leece BA, Lazar AC, et al. Synthesis and evaluation of hydrophilic linkers for antibody-maytansinoid conjugates. J Med Chem. 2011;54:3606–23. doi:10.1021/jm2002958.CrossRefPubMed

11.

Wang L, Amphlett G, Blattler WA, Lambert JM, Zhang W. Structural characterization of the maytansinoid-monoclonal antibody immunoconjugate, huN901-DM1, by mass spectrometry. Protein Sci. 2005;14:2436–46. doi:10.1110/ps.051478705.CrossRefPubMedPubMedCentral

12.

Junutula JR, Raab H, Clark S, Bhakta S, Leipold DD, Weir S, et al. Site-specific conjugation of a cytotoxic drug to an antibody improves the therapeutic index. Nat Biotechnol. 2008;26:925–32. doi:10.1038/nbt.1480.CrossRefPubMed

13.

Ricart AD. Antibody-drug conjugates of calicheamicin derivative: gemtuzumab ozogamicin and inotuzumab ozogamicin. Clin Cancer Res. 2011;17:6417–27. doi:10.1158/1078-0432.CCR-11-0486.CrossRefPubMed

14.

Bross PF, Beitz J, Chen G, Chen XH, Duffy E, Kieffer L, et al. Approval summary: gemtuzumab ozogamicin in relapsed acute myeloid leukemia. Clin Cancer Res. 2001;7:1490–6.PubMed

15.

Kim MT, Chen Y, Marhoul J, Jacobson F. Statistical modeling of the drug load distribution on trastuzumab emtansine (Kadcyla), a lysine-linked antibody drug conjugate. Bioconjug Chem. 2014;25:1223–32. doi:10.1021/bc5000109.CrossRefPubMed

16.

Doronina SO, Toki BE, Torgov MY, Mendelsohn BA, Cerveny CG, Chace DF, et al. Development of potent monoclonal antibody auristatin conjugates for cancer therapy. Nature Biotechnol. 2003;21:778–84. doi:10.1038/nbt832.CrossRef

17.

Sun MM, Beam KS, Cerveny CG, Hamblett KJ, Blackmore RS, Torgov MY, et al. Reduction-alkylation strategies for the modification of specific monoclonal antibody disulfides. Bioconjug Chem. 2005;16:1282–90. doi:10.1021/bc050201y.CrossRefPubMedPubMedCentral

18.

Shen BQ, Xu K, Liu L, Raab H, Bhakta S, Kenrick M, et al. Conjugation site modulates the in vivo stability and therapeutic activity of antibody-drug conjugates. Nature Biotechnol. 2012;30:184–9. doi:10.1038/nbt.2108.CrossRef

19.

Kalia D, Malekar PV, Parthasarathy M. Exocyclic olefinic maleimides: synthesis and application for stable and thiol-selective bioconjugation. Angew Chem Int Ed Engl. 2016;55:1432–5. doi:10.1002/anie.201508118.CrossRefPubMed

20.

Badescu G, Bryant P, Bird M, Henseleit K, Swierkosz J, Parekh V, et al. Bridging disulfides for stable and defined antibody drug conjugates. Bioconjug Chem. 2014;25:1124–36. doi:10.1021/bc500148x.CrossRefPubMed

21.

Jones MW, Strickland RA, Schumacher FF, Caddick S, Baker JR, Gibson MI, et al. Polymeric dibromomaleimides as extremely efficient disulfide bridging bioconjugation and pegylation agents. J Am Chem Soc. 2012;134:1847–52. doi:10.1021/ja210335f.CrossRefPubMed

22.

Maruani A, Smith ME, Miranda E, Chester KA, Chudasama V, Caddick S. A plug-and-play approach to antibody-based therapeutics via a chemoselective dual click strategy. Nat Commun. 2015;6:6645. doi:10.1038/ncomms7645.CrossRefPubMedPubMedCentral

23.

Lyons A, King DJ, Owens RJ, Yarranton GT, Millican A, Whittle NR, et al. Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. Protein Eng. 1990;3:703–8. doi:10.1093/protein/3.8.703.CrossRefPubMed

24.

Stimmel JB, Merrill BM, Kuyper LF, Moxham CP, Hutchins JT, Fling ME, et al. Site-specific conjugation on serine right-arrow cysteine variant monoclonal antibodies. J Biol Chem. 2000;275:30445–50. doi:10.1074/jbc.M001672200.CrossRefPubMed

25.

Sukumaran S, Gadkar K, Zhang C, Bhakta S, Liu L, Xu K, et al. Mechanism-based pharmacokinetic/pharmacodynamic model for THIOMAB drug conjugates. Pharm Res. 2015;32:1884–93. doi:10.1007/s11095-014-1582-1.CrossRefPubMed

26.

Casi G, Huguenin-Dezot N, Zuberbuhler K, Scheuermann J, Neri D. Site-specific traceless coupling of potent cytotoxic drugs to recombinant antibodies for pharmacodelivery. J Am Chem Soc. 2012;134:5887–92. doi:10.1021/ja211589m.CrossRefPubMed

27.

Zhang C, Welborn M, Zhu T, Yang NJ, Santos MS, Van Voorhis T, et al. pi-Clamp-mediated cysteine conjugation. Nat Chem. 2016;8:120–8. doi:10.1038/nchem.2413.PubMedPubMedCentral

28.

Wang L, Brock A, Herberich B, Schultz PG. Expanding the genetic code of Escherichia coli. Science. 2001;292:498–500. doi:10.1126/science.1060077.CrossRefPubMed

29.

Liu W, Brock A, Chen S, Chen S, Schultz PG. Genetic incorporation of unnatural amino acids into proteins in mammalian cells. Nat Methods. 2007;4:239–44. doi:10.1038/nmeth 1016.CrossRefPubMed

30.

Liu CC, Schultz PG. Adding new chemistries to the genetic code. Annu Rev Biochem. 2010;79:413–44. doi:10.1146/annurev.biochem.052308.105824.CrossRefPubMed

31.

Axup JY, Bajjuri KM, Ritland M, Hutchins BM, Kim CH, Kazane SA, et al. Synthesis of site-specific antibody-drug conjugates using unnatural amino acids. Proc Natl Acad Sci. 2012;109:16101–6. doi:10.1073/pnas.1211023109.CrossRefPubMedPubMedCentral

32.

Tian F, Lu Y, Manibusan A, Sellers A, Tran H, Sun Y, et al. A general approach to site-specific antibody drug conjugates. Proc Natl Acad Sci. 2014;111:1766–71. doi:10.1073/pnas.1321237111.CrossRefPubMedPubMedCentral

33.

Zimmerman ES, Heibeck TH, Gill A, Li X, Murray CJ, Madlansacay MR, et al. Production of site-specific antibody-drug conjugates using optimized non-natural amino acids in a cell-free expression system. Bioconjug Chem. 2014;25:351–61. doi:10.1021/bc400490z.CrossRefPubMed

34.

Jung ST, Kang TH, Kelton W, Georgiou G. Bypassing glycosylation: engineering aglycosylated full-length IgG antibodies for human therapy. Curr Opin Biotechnol. 2011;22:858–67. doi:10.1016/j.copbio.2011.03.002.CrossRefPubMed

35.

Ramakrishnan B, Qasba PK. Structure-based design of beta 1,4-galactosyltransferase I (beta 4Gal-T1) with equally efficient N-acetylgalactosaminyltransferase activity: point mutation broadens beta 4Gal-T1 donor specificity. J Biol Chem. 2002;277:20833–9. doi:10.1074/jbc.M111183200.CrossRefPubMed

36.

Boeggeman E, Ramakrishnan B, Pasek M, Manzoni M, Puri A, Loomis KH, et al. Site specific conjugation of fluoroprobes to the remodeled Fc N-glycans of monoclonal antibodies using mutant glycosyltransferases: application for cell surface antigen detection. Bioconjug Chem. 2009;20:1228–36. doi:10.1021/bc900103p.CrossRefPubMedPubMedCentral

37.

Zeglis BM, Davis CB, Aggeler R, Kang HC, Chen A, Agnew BJ, et al. Enzyme-mediated methodology for the site-specific radiolabeling of antibodies based on catalyst-free click chemistry. Bioconjug Chem. 2013;24:1057–67. doi:10.1021/bc400122c.CrossRefPubMedPubMedCentral

38.

Van DFL. Van Gr. Google Patents: WIJDEVEN MA; 2014.

39.

Zhou Q, Stefano JE, Manning C, Kyazike J, Chen B, Gianolio DA, et al. Site-specific antibody-drug conjugation through glycoengineering. Bioconjug Chem. 2014;25:510–20. doi:10.1021/bc400505q.CrossRefPubMed

40.

Ghaderi D, Taylor RE, Padler-Karavani V, Diaz S, Varki A. Implications of the presence of N-glycolylneuraminic acid in recombinant therapeutic glycoproteins. Nat Biotechnol. 2010;28:863–7. doi:10.1038/nbt.1651.CrossRefPubMedPubMedCentral

41.

Du J, Meledeo MA, Wang Z, Khanna HS, Paruchuri VD, Yarema KJ. Metabolic glycoengineering: sialic acid and beyond. Glycobiology. 2009;19:1382–401. doi:10.1093/glycob/cwp115.CrossRefPubMedPubMedCentral

42.

Jeger S, Zimmermann K, Blanc A, Grunberg J, Honer M, Hunziker P, et al. Site-specific and stoichiometric modification of antibodies by bacterial transglutaminase. Angew Chem Int Ed Engl. 2010;49:9995–7. doi:10.1002/anie.201004243.CrossRefPubMed

43.

Dennler P, Chiotellis A, Fischer E, Bregeon D, Belmant C, Gauthier L, et al. Transglutaminase-based chemo-enzymatic conjugation approach yields homogeneous antibody-drug conjugates. Bioconjug Chem. 2014;25:569–78. doi:10.1021/bc400574z.CrossRefPubMed

44.

Strop P, Liu SH, Dorywalska M, Delaria K, Dushin RG, Tran TT, et al. Location matters: site of conjugation modulates stability and pharmacokinetics of antibody drug conjugates. Chem Biol. 2013;20:161–7. doi:10.1016/j.chembiol.2013.01.010.CrossRefPubMed

45.

Farias SE, Strop P, Delaria K, Galindo Casas M, Dorywalska M, Shelton DL, et al. Mass spectrometric characterization of transglutaminase based site-specific antibody-drug conjugates. Bioconjug Chem. 2014;25:240–50. doi:10.1021/bc4003794.CrossRefPubMed

46.

Siegmund V, Schmelz S, Dickgiesser S, Beck J, Ebenig A, Fittler H, et al. Locked by design: A conformationally constrained transglutaminase tag enables efficient site-specific conjugation. Angew Chem Int Ed Engl. 2015;54:13420–4. doi:10.1002/anie.201504851.CrossRefPubMed

47.

Mao H, Hart SA, Schink A, Pollok BA. Sortase-mediated protein ligation: a new method for protein engineering. J Am Chem Soc. 2004;126:2670–1. doi:10.1021/ja039915e.CrossRefPubMed

48.

Popp MW, Antos JM, Grotenbreg GM, Spooner E, Ploegh HL. Sortagging: a versatile method for protein labeling. Nat Chem Biol. 2007;3:707–8. doi:10.1038/nchembio.2007.31.CrossRefPubMed

49.

Mohlmann S, Mahlert C, Greven S, Scholz P, Harrenga A. In vitro sortagging of an antibody fab fragment: overcoming unproductive reactions of sortase with water and lysine side chains. Chembiochem. 2011;12:1774–80. doi:10.1002/cbic.201100002.CrossRefPubMed

50.

Williamson DJ, Fascione MA, Webb ME, Turnbull WB. Efficient N-terminal labeling of proteins by use of sortase. Angew Chem Int Ed Engl. 2012;51:9377–80. doi:10.1002/anie.201204538.CrossRefPubMed

51.

Chen I, Dorr BM, Liu DR. A general strategy for the evolution of bond-forming enzymes using yeast display. Proc Natl Acad Sci. 2011;108:11399–404. doi:10.1073/pnas.1101046108.CrossRefPubMedPubMedCentral

52.

Beerli RR, Hell T, Merkel AS, Grawunder U. Sortase enzyme-mediated generation of site-specifically conjugated antibody drug conjugates with high In Vitro and In vivo potency. PLoS ONE. 2015;10:e0131177. doi:10.1371/journal.pone.0131177.CrossRefPubMedPubMedCentral

53.

Dierks T, Dickmanns A, Preusser-Kunze A, Schmidt B, Mariappan M, von Figura K, et al. Molecular basis for multiple sulfatase deficiency and mechanism for formylglycine generation of the human formylglycine-generating enzyme. Cell. 2005;121:541–52. doi:10.1016/j.cell.2005.03.001.CrossRefPubMed

54.

Carrico IS, Carlson BL, Bertozzi CR. Introducing genetically encoded aldehydes into proteins. Nat Chem Biol. 2007;3:321–2. doi:10.1038/nchembio878.CrossRefPubMed

55.

Agarwal P, van der Weijden J, Sletten EM, Rabuka D, Bertozzi CR. A pictet-spengler ligation for protein chemical modification. Proc Natl Acad Sci. 2013;110:46–51. doi:10.1073/pnas.1213186110.CrossRefPubMedPubMedCentral

56.

Agarwal P, Kudirka R, Albers AE, Barfield RM, de Hart GW, Drake PM, et al. Hydrazino-pictet-spengler ligation as a biocompatible method for the generation of stable protein conjugates. Bioconjug Chem. 2013;24:846–51. doi:10.1021/bc400042a.CrossRefPubMed

57.

Drake PM, Albers AE, Baker J, Banas S, Barfield RM, Bhat AS, et al. Aldehyde tag coupled with HIPS chemistry enables the production of ADCs conjugated site-specifically to different antibody regions with distinct in vivo efficacy and PK outcomes. Bioconjug Chem. 2014;25:1331–41. doi:10.1021/bc500189z.CrossRefPubMedPubMedCentral

58.

Holder PG, Jones LC, Drake PM, Barfield RM, Banas S, de Hart GW, et al. Reconstitution of formylglycine-generating enzyme with copper(II) for aldehyde tag conversion. J Biol Chem. 2015;290:15730–45. doi:10.1074/jbc.M115.652669.CrossRefPubMedPubMedCentral

59.

Schumacher D, Helma J, Mann FA, Pichler G, Natale F, Krause E, et al. Versatile and efficient site-specific protein functionalization by tubulin tyrosine ligase. Angew Chem Int Ed Engl. 2015;54:13787–91. doi:10.1002/anie.201505456.CrossRefPubMed

60.

Janke C The tubulin code: molecular components, readout mechanisms, and functions. J Cell Biol. 2014;206:461–72. doi:10.1083/jcb.201406055.CrossRefPubMedPubMedCentral

61.

Prota AE, Magiera MM, Kuijpers M, Bargsten K, Frey D, Wieser M, et al. Structural basis of tubulin tyrosination by tubulin tyrosine ligase. J Cell Biol. 2013;200:259–70. doi:10.1083/jcb.201211017.CrossRefPubMedPubMedCentral

62.

Lehar SM, Pillow T, Xu M, Staben L, Kajihara KK, Vandlen R, et al. Novel antibody-antibiotic conjugate eliminates intracellular S. aureus. Nature. 2015;527:323–8. doi:10.1038/nature16057.CrossRefPubMed

Title: Current Status: Site-Specific Antibody Drug Conjugates
Authors: Dominik Schumacher
Christian P. R. Hackenberger
Heinrich Leonhardt
Jonas Helma
Publication date: 01-05-2016
Publisher: Springer US
Published in: Journal of Clinical Immunology / Issue Special Issue 1/2016
Print ISSN: 0271-9142
Electronic ISSN: 1573-2592
DOI: https://doi.org/10.1007/s10875-016-0265-6

ACC 2024 Congress Coverage

Heart Failure Video

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Current Status: Site-Specific Antibody Drug Conjugates

Abstract

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Special Issue 1/2016

Metabolic Control of Plasma Cell Differentiation- What We Know and What We Don't Know

Autophagy in Plasma Cell Ontogeny and Malignancy

Translating Inhibitory Fc Receptor Biology into Novel Therapeutic Approaches

Erratum to: The Elements Steering Pathogenesis in IgG-Mediated Alloimmune Diseases

The FcγR/IgG Interaction as Target for the Treatment of Autoimmune Diseases

Dissecting Epigenetic Dysregulation of Primary Antibody Deficiencies

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page