Bivalent EGFR-Targeting DARPin-MMAE Conjugates
Karsten L, Janson N, Le Joncour V, Alam S, Müller B, Ramanathan JT, Laakkonen P, Sewald N, Müller K (2022)
International Journal of Molecular Sciences 23(5): 2468.
Zeitschriftenaufsatz
| Veröffentlicht | Englisch
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ijms-23-02468-v2.pdf
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Autor*in
Karsten, LennardUniBi;
Janson, NilsUniBi;
Le Joncour, Vadim;
Alam, SarfarazUniBi;
Müller, BenjaminUniBi;
Ramanathan, Jayendrakishore Tanjore;
Laakkonen, Pirjo;
Sewald, NorbertUniBi ;
Müller, KristianUniBi
Einrichtung
Abstract / Bemerkung
Epidermal growth factor receptor (EGFR) is a validated tumor marker overexpressed in various cancers such as squamous cell carcinoma (SSC) of the head and neck and gliomas. We constructed protein-drug conjugates based on the anti-EGFR designed ankyrin repeat protein (DARPin) E01, and compared the bivalent DARPin dimer (DD1) and a DARPin-Fc (DFc) to the monomeric DARPin (DM) and the antibody derived scFv425-Fc (scFvFc) in cell culture and a mouse model. The modular conjugation system, which was successfully applied for the preparation of protein-drug and -dye conjugates, uses bio-orthogonal protein-aldehyde generation by the formylglycine-generating enzyme (FGE). The generated carbonyl moiety is addressed by a bifunctional linker with a pyrazolone for a tandem Knoevenagel reaction and an azide for strain-promoted azide-alkyne cycloaddition (SPAAC). The latter reaction with a PEGylated linker containing a dibenzocyclooctyne (DBCO) for SPAAC and monomethyl auristatin E (MMAE) as the toxin provided the stable conjugates DD1-MMAE (drug-antibody ratio, DAR = 2.0) and DFc-MMAE (DAR = 4.0) with sub-nanomolar cytotoxicity against the human squamous carcinoma derived A431 cells. In vivo imaging of Alexa Fluor 647-dye conjugates in A431-xenografted mice bearing subcutaneous tumors as the SCC model revealed unspecific binding of bivalent DARPins to the ubiquitously expressed EGFR. Tumor-targeting was verified 6 h post-injection solely for DD1 and scFvFc. The total of four administrations of 6.5 mg/kg DD1-MMAE or DFc-MMAE twice weekly did not cause any sequela in mice. MMAE conjugates showed no significant anti-tumor efficacy in vivo, but a trend towards increased necrotic areas (p = 0.2213) was observed for the DD1-MMAE (n = 5).
Stichworte
EGFR;
DARPin;
antibody-drug conjugates;
MMAE conjugates;
Knoevenagel ligation;
formylglycine-generating enzyme;
cytotoxicity;
in vivo imaging;
xenograft;
SCC model
Erscheinungsjahr
2022
Zeitschriftentitel
International Journal of Molecular Sciences
Band
23
Ausgabe
5
Art.-Nr.
2468
Urheberrecht / Lizenzen
eISSN
1422-0067
Finanzierungs-Informationen
Open-Access-Publikationskosten wurden durch die Universität Bielefeld gefördert.
Page URI
https://pub.uni-bielefeld.de/record/2961406
Zitieren
Karsten L, Janson N, Le Joncour V, et al. Bivalent EGFR-Targeting DARPin-MMAE Conjugates. International Journal of Molecular Sciences. 2022;23(5): 2468.
Karsten, L., Janson, N., Le Joncour, V., Alam, S., Müller, B., Ramanathan, J. T., Laakkonen, P., et al. (2022). Bivalent EGFR-Targeting DARPin-MMAE Conjugates. International Journal of Molecular Sciences, 23(5), 2468. https://doi.org/10.3390/ijms23052468
Karsten, Lennard, Janson, Nils, Le Joncour, Vadim, Alam, Sarfaraz, Müller, Benjamin, Ramanathan, Jayendrakishore Tanjore, Laakkonen, Pirjo, Sewald, Norbert, and Müller, Kristian. 2022. “Bivalent EGFR-Targeting DARPin-MMAE Conjugates”. International Journal of Molecular Sciences 23 (5): 2468.
Karsten, L., Janson, N., Le Joncour, V., Alam, S., Müller, B., Ramanathan, J. T., Laakkonen, P., Sewald, N., and Müller, K. (2022). Bivalent EGFR-Targeting DARPin-MMAE Conjugates. International Journal of Molecular Sciences 23:2468.
Karsten, L., et al., 2022. Bivalent EGFR-Targeting DARPin-MMAE Conjugates. International Journal of Molecular Sciences, 23(5): 2468.
L. Karsten, et al., “Bivalent EGFR-Targeting DARPin-MMAE Conjugates”, International Journal of Molecular Sciences, vol. 23, 2022, : 2468.
Karsten, L., Janson, N., Le Joncour, V., Alam, S., Müller, B., Ramanathan, J.T., Laakkonen, P., Sewald, N., Müller, K.: Bivalent EGFR-Targeting DARPin-MMAE Conjugates. International Journal of Molecular Sciences. 23, : 2468 (2022).
Karsten, Lennard, Janson, Nils, Le Joncour, Vadim, Alam, Sarfaraz, Müller, Benjamin, Ramanathan, Jayendrakishore Tanjore, Laakkonen, Pirjo, Sewald, Norbert, and Müller, Kristian. “Bivalent EGFR-Targeting DARPin-MMAE Conjugates”. International Journal of Molecular Sciences 23.5 (2022): 2468.
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