Metabolomic isotopic tracing can provide flux information useful for understanding drug mechanisms. For that, NMR has the unique advantage of giving positional isotope enrichment information, but the current ¹³C 1D NMR approach suffers from low sensitivity and high overlaps. We developed a new 2D heteronuclear NMR experiment incorporating J-scaling and distortion-free elements that allows for quantitative analysis of multiplets with high sensitivity and resolution. When applied to an old chemotherapeutic drug, the approach provided a quantitative estimation of TCA-cycle turns, confirming the conventional mechanism of its mitochondrial metabolic enhancement. Additionally, the approach identified a new mechanism of the higher contribution of the pentose phosphate pathway to serine synthesis in the cytosolic compartment, possibly explaining the broad pharmacological activities of the drug. Our approach may prove beneficial in helping to find new usages or metabolic mechanisms of other drugs.

Two new pyrrolosesquiterpenes, glaciapyrroles D (1) and E (2) were discovered along with the previously reported glaciapyrrole A (3) from Streptomyces sp. GGS53 strain isolated from deep-sea sediment. This study elucidated the planar structures of 1 and 2 using nuclear magnetic resonance (NMR), mass spectrometry (MS), ultraviolet (UV), and infrared (IR) spectroscopic data. The absolute configurations of the glaciapyrroles were determined by Mosher’s method, circular dichroism spectroscopy, and X-ray crystallography. Under 366 nm UV irradiation, the glaciapyrroles were systematically converted to the corresponding photoglaciapyrroles (4–6) via photoisomerization, resulting in the diversification of the glaciapyrrole family compounds. The transformation of the glaciapyrrole Z to E isomers occurred in a 1:1 ratio, based on virtual validation of the photoisomerization of these olefinic compounds by ¹H-NMR spectroscopy and liquid chromatography/mass spectrometry (LC/MS) analysis. Finally, when encapsulated in poly(lactic-co-glycolic acid) nanoparticles, glaciapyrrole E and photoglaciapyrrole E displayed significant inhibitory activity against influenza A virus. This is the first report of antiviral effects from glaciapyrrole family compounds, whose biological functions have only been subjected to limited studies so far.

Phenylpropanoids are naturally occurring secondary metabolites that exhibit various biological activities such as ultra-violet (UV) light protection and reactive-oxygen species (ROS) scavenging. In this study, we utilized a light-emitting diode (LED) based in-situ UV irradiation nuclear magnetic resonance (NMR) technique to monitor the photoisomerization reactions of these phenylpropanoids under UV irradiation in real-time. Through this approach, we measured the photochemical reaction rates and photostationary state (PSS) ratios of these molecules and observed distinct reaction rate and PSS ratio information depending on the variation of substituent groups in each phenylpropanoid molecule. We also evaluated the radical scavenging activity (RSA) for each photochemical product through diphenyl-1-picrylhydrazyl radical (DPPH) assay and 2,2’-azino-bis(3-ethylbenzenthiazoline-6-sulphonic acid) (ABTS) assay. We found that the photoisomerization product of caffeic acid can increase both DPPH and ABTS radical scavenging activities, and confirmed the enhanced ABTS radical scavenging ability of caffeic acid cis-isomer based on the online high-pressure liquid chromatography (HPLC)-ABTS analysis and the PSS ratio information of each isomer.

Complex nuclear magnetic resonance (NMR) signals of organic compounds containing multiple analogous substructures or mixtures pose a significant challenge to structural identification, thus resulting in frequent misassignment of structures. The GEMSTONE method, a single‐scan technique that selectively excites a specific proton signal among the crowded NMR signals, was recently proposed as a solution. However, its extension to the polarization transfer method for heteronuclear spin systems was unsuccessful. Herein, we present an extension method that addresses the altered heteronuclear polarization transfer efficiency and enables the acquisition of ultraselective ¹³C and ¹H‐¹³C correlation NMR subspectra with hertz‐level signal selectivity in both dimensions. We demonstrate the effectiveness of this technique in the structural analysis of a chromopeptide pharmaceutical and a diastereomeric mixture of a fungicide.