Šoltésová Mária, Pinon Arthur C, Aussenac Fabien, Schlagnitweit Judith, Reiter Christian, Purea Armin, Melzi Roberto, Engelke Frank, Martin Dave, Krambeck Stefanie, Biscans Annabelle, Kay Emma, Emsley Lyndon, Schantz Staffan
Swedish NMR Centre, University of Gothenburg, 413 90 Gothenburg, Sweden.
Swedish NMR Centre, University of Gothenburg, 413 90 Gothenburg, Sweden.
J Magn Reson. 2025 Feb;371:107827. doi: 10.1016/j.jmr.2024.107827. Epub 2024 Dec 27.
A new 3.2 mm H-F-X magic angle spinning dynamic nuclear polarization NMR (MAS DNP-NMR) probe was developed with a unique coil design with separate radiofrequency channels for H excitation and C or F detection to enable acquisition of H-F cross-polarization (CP) MAS experiments, direct-detected F spectra with proton decoupling, and acquisition on C with simultaneous double decoupling on the H and 19F channels as well as H-F-C double-CP experiments under low temperature MAS DNP conditions. We use these sequences to study AZD2811, which is an active pharmaceutical ingredient (API), in its pure dry state as well as in its corresponding drug delivery formulation consisting of drug-loaded polymeric nanoparticles (PNPs). Included in this study are also small interfering RNAs (siRNAs) for therapeutic targeting of peptidyl-prolyl cis-trans isomerase B (Ppib) mRNA. We demonstrate that H-F CP MAS experiments performed on the new HFX probe represent a notable advantage over usually acquired direct-detected F experiments. The indirect F DNP enhancement ε(F) = 26 was obtained via H-F CP for the pure API impregnated with DNP solution, with an overall 30-fold sensitivity gain compared to the direct-detected F experiment under similar conditions. DNP enhancement value of ε(F) = 42 was obtained via H-F CP for the polymeric nanoparticle suspension and ε(F) ≈ 150 were obtained for two different siRNAs in frozen DNP solution.
开发了一种新型的3.2毫米H-F-X魔角旋转动态核极化核磁共振（MAS DNP-NMR）探头，其独特的线圈设计具有用于H激发和C或F检测的独立射频通道，能够进行H-F交叉极化（CP）MAS实验、带质子去耦的直接检测F谱，以及在低温MAS DNP条件下在C上进行H和19F通道同时双去耦的采集以及H-F-C双CP实验。我们使用这些序列来研究处于纯干燥状态的活性药物成分（API）AZD2811，以及由载药聚合物纳米颗粒（PNP）组成的相应药物递送制剂。本研究还包括用于治疗性靶向肽基脯氨酰顺反异构酶B（Ppib）mRNA的小干扰RNA（siRNA）。我们证明，在新的HFX探头上进行的H-F CP MAS实验相对于通常获得的直接检测F实验具有显著优势。对于浸渍有DNP溶液的纯API，通过H-F CP获得间接F DNP增强ε(F)=26，与在类似条件下的直接检测F实验相比，整体灵敏度提高了30倍。对于聚合物纳米颗粒悬浮液，通过H-F CP获得的DNP增强值ε(F)=42，对于冷冻DNP溶液中的两种不同siRNA，ε(F)≈150。
