Synthesis and Application of Two-Photon Active Fluorescent Rhodol Dyes for Antibody Conjugation and In Vitro Cell Imaging

Dénes Szepesi Kovács; Balázs Chiovini; Dalma Müller; Estilla Zsófia Tóth; Anna Fülöp; Péter Ábrányi-Balogh; Lucia Wittner; György Várady; Ödön Farkas; Gábor Turczel; Gergely Katona; Balázs Győrffy; György Miklós Keserű; Zoltán Mucsi; Balázs J Rózsa; Ervin Kovács

doi:10.1021/acsomega.3c01796

Synthesis and Application of Two-Photon Active Fluorescent Rhodol Dyes for Antibody Conjugation and In Vitro Cell Imaging

ACS Omega. 2023 Jun 14;8(25):22836-22843. doi: 10.1021/acsomega.3c01796. eCollection 2023 Jun 27.

Authors

Dénes Szepesi Kovács^{1

2

3}, Balázs Chiovini⁴, Dalma Müller^{5

6

7}, Estilla Zsófia Tóth^{3

7

8}, Anna Fülöp⁹, Péter Ábrányi-Balogh^{1

2

3}, Lucia Wittner^{3

8}, György Várady¹⁰, Ödön Farkas¹¹, Gábor Turczel¹², Gergely Katona⁴, Balázs Győrffy^{3

5

6

13}, György Miklós Keserű^{1

2

3}, Zoltán Mucsi^{14

15}, Balázs J Rózsa^{4

14

16}, Ervin Kovács^{9

17}

Affiliations

¹ Medicinal Chemistry Research Group, Research Centre for Natural Sciences, H-1117 Budapest, Hungary.
² Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, H-1111 Budapest, Hungary.
³ National Laboratory for Drug Research and Development, H-1117 Budapest, Hungary.
⁴ Faculty of Information Technology and Bionics, Pázmány Péter Catholic University, H-1444 Budapest, Hungary.
⁵ Oncology Biomarker Research Group, Research Centre for Natural Sciences, H-1117 Budapest, Hungary.
⁶ Department of Bioinformatics, Semmelweis University, H-1094 Budapest, Hungary.
⁷ Semmelweis University Doctoral School, H-1085 Budapest Hungary.
⁸ Integrative Neuroscience Research Group, Research Centre for Natural Sciences, H-1117 Budapest, Hungary.
⁹ Femtonics Ltd., H-1094 Budapest, Hungary.
¹⁰ Molecular Cell Biology Research Group, Research Centre for Natural Sciences, H-1117 Budapest, Hungary.
¹¹ Department of Organic Chemistry, Eötvös Loránd University, H-1117 Budapest, Hungary.
¹² NMR Research Laboratory, Research Centre for Natural Sciences, H-1117 Budapest, Hungary.
¹³ Department of Pediatrics, Semmelweis University, H-1094 Budapest, Hungary.
¹⁴ Brain Vision Center, H-1094 Budapest, Hungary.
¹⁵ Faculty of Materials and Chemical Sciences, University of Miskolc, Miskolc H-3515, Hungary.
¹⁶ Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, H-1083 Budapest, Hungary.
¹⁷ Polymer Chemistry and Physics Research Group, Research Centre for Natural Sciences, H-1117 Budapest, Hungary.

Abstract

A novel family of julolidine-containing fluorescent rhodols equipped with a wide variety of substituents was synthesized in a versatile two-step process. The prepared compounds were fully characterized and exhibited excellent fluorescence properties for microscopy imaging. The best candidate was conjugated to the therapeutic antibody trastuzumab through a copper-free strain-promoted azide-alkyne click reaction. The rhodol-labeled antibody was successfully applied for in vitro confocal and two-photon microscopy imaging of Her2+ cells.