Characterization of Aptamer BC 007 Substance and Product Using Circular Dichroism and Nuclear Magnetic Resonance Spectroscopy
| Autor: | Johannes Müller, Katrin Wenzel, Sarah Schulze-Rothe, Heike Nikolenko, Niels-Peter Becker, Margitta Dathe, Hanna Davideit, Peter Göttel, Annekathrin Haberland, G. Susan Srivatsa, Hardy Weisshoff |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification Circular dichroism Aqueous solution Magnetic Resonance Spectroscopy 030102 biochemistry & molecular biology Chemistry Aptamer Circular Dichroism Pharmaceutical Science Nuclear magnetic resonance spectroscopy Aptamers Nucleotide Nucleic Acid Denaturation Small molecule G-Quadruplexes 03 medical and health sciences Crystallography 030104 developmental biology Drug Stability Pharmaceutical Preparations Nucleic Acid Conformation Thermodynamics Denaturation (biochemistry) Nucleotide Spectroscopy |
| Zdroj: | Journal of pharmaceutical sciences. 107(8) |
| ISSN: | 1520-6017 |
| Popis: | Possible unwanted folding of biopharmaceuticals during manufacturing and storage has resulted in analysis schemes compared to small molecules that include bioanalytical characterization besides chemical characterization. Whether bioanalytical characterization is required for nucleotide-based drugs, may be decided on a case-by-case basis. Nucleotide-based pharmaceuticals, if chemically synthesized, occupy an intermediate position between small-molecule drugs and biologics. Here, we tested whether a physicochemical characterization of a nucleotide-based drug substance, BC 007, was adequate, using circular dichroism (CD) spectroscopy. Nuclear magnetic resonance confirmed CD data in one experimental setup. BC 007 forms a quadruplex structure under specific external conditions, which was characterized for its stability and structural appearance also after denaturation using CD and nuclear magnetic resonance. The amount of the free energy (ΔG 0 ) involved in quadruplex formation of BC 007 was estimated at +8.7 kJ/mol when dissolved in water and +1.4 kJ/mol in 154 mM NaCl, indicating structural instability under these conditions. However, dissolution of the substance in 5 mM of KCl reduced the ΔG 0 to −5.6 kJ/mol due to the stabilizing effect of cations. These results show that positive ΔG 0 of quadruplex structure formation in water and aqueous NaCl prevents BC 007 from preforming stable 3-dimensional structures, which could potentially affect drug function. |
| Databáze: | OpenAIRE |
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