Near UV and Visible Light-Induced Degradation of Bovine Serum Albumin and a Monoclonal Antibody Mediated by Citrate Buffer and Fe(III): Reduction vs Oxidation Pathways.

Light exposure during manufacturing, storage, and administration can lead to the photodegradation of therapeutic proteins. This photodegradation can be promoted by pharmaceutical buffers or impurities. Our laboratory has previously demonstrated that citrate-Fe(III) complexes generate the •CO2- radical anion when photoirradiated under near UV (λ = 320-400 nm) and visible light (λ = 400-800 nm) [Subelzu, N.; Schöneich, C. Mol. Pharmaceutics 2020, 17 (11), 4163-4179; Zhang, Y. Mol. Pharmaceutics 2022, 19 (11), 4026-4042]. Here, we evaluated the impact of citrate-Fe(III) on the photostability and degradation mechanisms of disulfide-containing proteins (bovine serum albumin (BSA) and NISTmAb) under pharmaceutically relevant conditions. We monitored and localized competitive disulfide reduction and protein oxidation by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analysis depending on the reaction conditions. These competitive pathways were affected by multiple factors, including light dose, Fe(III) concentration, protein concentration, the presence of oxygen, and light intensity.

Identification and Development of Cyclic Peptide Inhibitors of Hypoxia Inducible Factors 1 and 2 That Disrupt Hypoxia-Response Signaling in Cancer Cells.

Hypoxia inducible factor (HIF) is a heterodimeric transcription factor composed of an oxygen-regulated α subunit and a constitutively expressed ß subunit that serves as the master regulator of the cellular response to low oxygen concentrations. The HIF transcription factor senses and responds to hypoxia by significantly altering transcription and reprogramming cells to enable adaptation to a hypoxic microenvironment. Given the central role played by HIF in the survival and growth of tumors in hypoxia, inhibition of this transcription factor serves as a potential therapeutic approach for treating a variety of cancers. Here, we report the identification, optimization, and characterization of a series of cyclic peptides that disrupt the function of HIF-1 and HIF-2 transcription factors by inhibiting the interaction of both HIF-1α and HIF-2α with HIF-1ß. These compounds are shown to bind to HIF-α and disrupt the protein-protein interaction between the α and ß subunits of the transcription factor, resulting in disruption of hypoxia-response signaling by our lead molecule in several cancer cell lines.

Evoking stress reactivity in virtual reality: A systematic review and meta-analysis.

BACKGROUND: Virtual reality (VR) research probes stress environments that are infeasible to create in the real world. However, because research simulations are applied to narrow populations, it remains unclear if VR simulations can stimulate a broadly applicable stress-response. This systematic review and meta-analysis was conducted on studies using VR stress tasks and biomarkers. METHODS: Included papers (N = 52) measured cortisol, heart rate (HR), galvanic skin response (GSR), systolic blood pressure (SBP), diastolic blood pressure (DBP), respiratory sinus arrhythmia (RSA), parasympathetic activity (RMSSD), sympathovagal balance (LF/HF), and/or salivary alpha-amylase (sAA). Effect sizes (ES) and confidence intervals (CI) were calculated based on standardized mean change of baseline-to-peak biomarker levels. RESULTS: From baseline-to-peak (ES, CI), analyses showed a statistically significant change in cortisol (0.56, 0.28-0.83), HR (0.68, 0.53-0.82), GSR (0.59, 0.36-0.82), SBP (.55, 0.19-0.90), DBP (.64, 0.23-1.05), RSA (-0.59, -0.88 to -0.30), and sAA (0.27, 0.092-0.45). There was no effect for RMSSD and LF/HF. CONCLUSION: VR stress tasks elicited a varied magnitude of physiological stress reactivity. VR may be an effective tool in stress research.

Development of a Cyclic Peptide Inhibitor of the p6/UEV Protein-Protein Interaction.

The budding of HIV from infected cells is driven by the protein-protein interaction between the p6 domain of the HIV Gag protein and the UEV domain of the human TSG101 protein. We report the development of a cyclic peptide inhibitor of the p6/UEV interaction, from a non cell-permeable parent that was identified in a SICLOPPS screen. Amino acids critical for the activity of the parent cyclic peptide were uncovered using alanine-scanning, and a series of non-natural analogues synthesized and assessed. The most potent molecule disrupts the p6/UEV interaction with an IC50 of 6.17 ± 0.24 µM by binding to UEV with a Kd of 11.9 ± 2.8 µM. This compound is cell permeable and active in a cellular virus-like particle budding assay with an IC50 of ∼2 µM. This work further demonstrates the relative simplicity with which the potency and activity of cyclic peptides identified from SICLOPPS libraries can be optimized.