TY - JOUR

T1 - Antisense oligonucleotide gapmers containing phosphoryl guanidine groups reverse MDR1-mediated multiple drug resistance of tumor cells

AU - Kupryushkin, Maxim S.

AU - Filatov, Anton V.

AU - Mironova, Nadezhda L.

AU - Patutina, Olga A.

AU - Chernikov, Ivan V.

AU - Chernolovskaya, Elena L.

AU - Zenkova, Marina A.

AU - Pyshnyi, Dmitrii V.

AU - Stetsenko, Dmitry A.

AU - Altman, Sidney

AU - Vlassov, Valentin V.

N1 - Funding Information: The study was initiated in the framework of Russian Government Support for Research Projects implemented under supervision of world's leading scientists (agreement no. 14.B25.31.0028 with S.A. as the leading scientist). This work was funded by the Russian Science Foundation (grant no. 19-74-30011 ), the Russian Foundation for Basic Research (grant nos. 18-515-57006 , 18-29-09045 , and 18-29-08062 to D.A.S.), and the Russian Government-funded budget project of ICBFM SB RAS no. 121031300042-1 . The authors thank Ms A. Vladimirova for cell maintenance and Dr. O. Markov for the help with the analysis of flow cytometry data. Publisher Copyright: © 2021 Institute of Chemical Biology and Fundamental Medicine SB RAS

PY - 2022/3/8

Y1 - 2022/3/8

N2 - Antisense gapmer oligonucleotides containing phosphoryl guanidine (PG) groups, e.g., 1,3-dimethylimidazolidin-2-imine, at three to five internucleotidic positions adjacent to the 3′ and 5′ ends were prepared via the Staudinger chemistry, which is compatible with conditions of standard automated solid-phase phosphoramidite synthesis for phosphodiester and, notably, phosphorothioate linkages, and allows one to design a variety of gapmeric structures with alternating linkages, and deoxyribose or 2′-O-methylribose backbone. PG modifications increased nuclease resistance in serum-containing medium for more than 21 days. Replacing two internucleotidic phosphates by PG groups in phosphorothioate-modified oligonucleotides did not decrease their cellular uptake in the absence of lipid carriers. Increasing the number of PG groups from two to seven per oligonucleotide reduced their ability to enter the cells in the carrier-free mode. Cationic liposomes provided similar delivery efficiency of both partially PG-modified and unmodified oligonucleotides. PG-gapmers were designed containing three to four PG groups at both wings and a central “window” of seven deoxynucleotides with either phosphodiester or phosphorothioate linkages targeted to MDR1 mRNA providing multiple drug resistance of tumor cells. Gapmers efficiently silenced MDR1 mRNA and restored the sensitivity of tumor cells to chemotherapeutics. Thus, PG-gapmers can be considered as novel, promising types of antisense oligonucleotides for targeting biologically relevant RNAs.

AB - Antisense gapmer oligonucleotides containing phosphoryl guanidine (PG) groups, e.g., 1,3-dimethylimidazolidin-2-imine, at three to five internucleotidic positions adjacent to the 3′ and 5′ ends were prepared via the Staudinger chemistry, which is compatible with conditions of standard automated solid-phase phosphoramidite synthesis for phosphodiester and, notably, phosphorothioate linkages, and allows one to design a variety of gapmeric structures with alternating linkages, and deoxyribose or 2′-O-methylribose backbone. PG modifications increased nuclease resistance in serum-containing medium for more than 21 days. Replacing two internucleotidic phosphates by PG groups in phosphorothioate-modified oligonucleotides did not decrease their cellular uptake in the absence of lipid carriers. Increasing the number of PG groups from two to seven per oligonucleotide reduced their ability to enter the cells in the carrier-free mode. Cationic liposomes provided similar delivery efficiency of both partially PG-modified and unmodified oligonucleotides. PG-gapmers were designed containing three to four PG groups at both wings and a central “window” of seven deoxynucleotides with either phosphodiester or phosphorothioate linkages targeted to MDR1 mRNA providing multiple drug resistance of tumor cells. Gapmers efficiently silenced MDR1 mRNA and restored the sensitivity of tumor cells to chemotherapeutics. Thus, PG-gapmers can be considered as novel, promising types of antisense oligonucleotides for targeting biologically relevant RNAs.

KW - antisense oligonucleotide gapmer

KW - gene silencing

KW - intracellular accumulation

KW - MDR1

KW - nuclease resistance

KW - phosphoryl guanidine

KW - RNase H

UR - http://www.scopus.com/inward/record.url?scp=85121217937&partnerID=8YFLogxK

U2 - 10.1016/j.omtn.2021.11.025

DO - 10.1016/j.omtn.2021.11.025

M3 - Article

C2 - 34976439

AN - SCOPUS:85121217937

VL - 27

SP - 211

EP - 226

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

SN - 2162-2531

ER -