Photo‐affinity and Metabolic Labeling Probes Based on the Opioid Alkaloids

Abstract

The opioids are powerful analgesics but carry many contingencies that can lead to opioid‐use disorder. Chemical probes derived from the opioid alkaloids can provide deeper insight into the molecular interactions in a cellular context. Here, we designed and developed photo‐click morphine (PCM‐2) as a photo‐affinity probe based on morphine and dialkynyl‐acetyl morphine (DAAM) as a metabolic acetate reporter based on heroin. Application of these probes to SH‐SY5Y, HEK293T, and U2OS cells revealed that PCM‐2 and DAAM primarily localize to the lysosome amongst other locations inside the cell by confocal microscopy and chemical proteomics. Interaction site identification by mass spectrometry revealed the mitochondrial phosphate carrier protein, solute carrier family 25 member 3, SLC25A3, and histone H2B as acylation targets of DAAM. These data illustrate the utility of chemical probes to measure localization and protein interactions in a cellular context and will inform the design of probes based on the opioids in the future.

Evaluation of Antibacterial Functionalized Dihydropyrimidine Photoaffinity Probes Toward Mechanism of Action Studies

“…To explore the antibacterial mechanism, we considered that photoaffinity labeling (PAL) may allow us to identify specific biological targets. − Trifluoromethyl aryl diazirines can be ideal photolabile functional groups, , which could be used with the cellular lysate, or in pulse-chase experiments with live cells to label targets in their appropriate biological context. , As many of our most successful antibacterial compounds had multiple arenes as components ( 7a – j ), we considered that a trifluoromethyl diazirine could be placed on either aryl position, with our initial emphasis on the β-keto ester, as this arene appeared to have less impact on the antibacterial activity. We envisioned accessing 4-trifluoromethyldiaziryl substituted β-keto ester 11 from precursor acetophenone 12 via base-promoted Claisen-type condensation with diethyl carbonate (Figure ).…”