Download file (1.22 MB)
Download as zip (1.14 MB)

Collection

Citation

Description:

Oligonucleotides are short nucleic acids that serve as one of the most promising classes of drug modality. However, attempts to establish a physicochemical evaluation platform of oligonucleotides for acquiring a comprehensive view of their properties have been limited. As the chemical stability and the efficacy as well as the solution properties at a high concentration should be related to their higher-order structure and intra-/intermolecular interactions, their detailed understanding enables effective formulation development. Here, the higher-order structure and the thermodynamic stability of the thrombin-binding aptamer (TBA) and four modified TBAs, which have similar sequences but were expected to have different higher-order structures, were evaluated using ultraviolet spectroscopy (UV), circular dichroism (CD), differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR). Then, the relationship between the higher-order structure and the solution properties including solubility, viscosity, and stability was investigated. The impact of the higher-order structure on the antithrombin activity was also confirmed. The higher-order structure and intra-/intermolecular interactions of the oligonucleotides were affected by types of buffers because of different potassium concentrations, which are crucial for the formation of the G-quadruplex structure. Consequently, solution properties, such as solubility and viscosity, chemical stability, and antithrombin activity, were also influenced. Each instrumental analysis had a complemental role in investigating the higher-order structure of TBA and modified TBAs. The utility of each physicochemical characterization method during the preclinical developmental stages is also discussed.

Rights:

• In Copyright
  

  This document is the Accepted Manuscript version of a Published Work that appeared in final form in Molecular Pharmaceutics, copyright © 2024 American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.molpharmaceut.4c00177

Keyword: oligonucleotide, higher-order structure, thrombin aptamer, G-quadruplex, physicochemical profiling

Date published: 2024-07-01

Publisher: American Chemical Society (ACS)

Journal:

• Molecular Pharmaceutics (ISSN: 15438392) vol. 21 issue. 7 p. 3471-3484

Funding:

• Eisai Co., Ltd.
• University of Tsukuba
• Takeda Pharmaceutical Company
• Astellas Pharma
• National Institute for Materials Science
• Sawai Pharmaceutical
• Materials Open Platform for Pharmaceutical Science
• Mitsubishi Tanabe Pharma Corporation
• Daiichi-Sankyo

Manuscript type: Author's version (Accepted manuscript)

MDR DOI: https://doi.org/10.48505/nims.4821

First published URL: https://doi.org/10.1021/acs.molpharmaceut.4c00177

Related item:

Other identifier(s):

Contact agent:

Updated at: 2025-06-15 08:30:25 +0900

Published on MDR: 2025-06-15 08:20:14 +0900