Discovery of Bivalent GalNAc-Conjugated Betulin as a Potent ASGPR-Directed Agent against Hepatocellular Carcinoma

Emil Yu Yamansarov, Elena V. Lopatukhina, Sergei A. Evteev, Dmitry A. Skvortsov, Anton V. Lopukhov, Sergey V. Kovalev, Alexander N. Vaneev, Dmitry O. Shkil', Roman A. Akasov, Alexander N. Lobov, Victor A. Naumenko, Ekaterina N. Pavlova, Oxana O. Ryabaya, Olga Yu Burenina, Yan A. Ivanenkov, Natalia L. Klyachko, Alexander S. Erofeev, Petr V. Gorelkin, Elena K. Beloglazkina, Alexander G. Majouga

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


Herein, we describe the design, synthesis, and biological evaluation of novel betulin and N-acetyl-d-galactosamine (GalNAc) glycoconjugates and suggest them as targeted agents against hepatocellular carcinoma. We prepared six conjugates derived via the C-3 and C-28 positions of betulin with one or two saccharide ligands. These molecules demonstrate high affinity to the asialoglycoprotein receptor (ASGPR) of hepatocytes assessed by in silico modeling and surface plasmon resonance tests. Cytotoxicity studies in vitro revealed a bivalent conjugate with moderate activity, selectivity of action, and cytostatic properties against hepatocellular carcinoma cells HepG2. An additional investigation confirmed the specific engagement with HepG2 cells by the enhanced generation of reactive oxygen species. Stability tests demonstrated its lability to acidic media and to intracellular enzymes. Therefore, the selected bivalent conjugate represents a new potential agent targeted against hepatocellular carcinoma. Further extensive studies of the cellular uptake in vitro and the real-time microdistribution in the murine liver in vivo for fluorescent dye-labeled analogue showed its selective internalization into hepatocytes due to the presence of GalNAc ligand in comparison with reference compounds. The betulin and GalNAc glycoconjugates can therefore be considered as a new strategy for developing therapeutic agents based on natural triterpenoids.

Original languageEnglish
Pages (from-to)763-781
Number of pages19
JournalBioconjugate Chemistry
Issue number4
Publication statusPublished - 21 Apr 2021


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