Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Myeloperoxidase as a therapeutic target for oxidative damage in Alzheimer’s disease

doi: 10.1080/14756366.2025.2456282

Figure Lengend Snippet: Chemical structures of myeloperoxidase inhibitors. The chemical structures of the aromatic hydroxamates SHA, 5-ASA, tryptamine, 4-ABAH, AZD4831, AZD5904 and AZD3241 are shown. These compounds demonstrate different mechanisms of action against MPO (reversible inhibition, accumulation of compound II and irreversible inhibition). Figure created using Biorender software.

Article Snippet: In this sense, AstraZeneca has focused considerable attention on obtaining MPO inhibitors to treat different pathologies, with the aim of reducing the damage caused by HOCl.

Techniques: Inhibition, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: DNA-based fluorescent probes of NOS2 activity in live brains

doi: 10.1073/pnas.2003034117

Figure Lengend Snippet: NOckoutFn detects NO in mouse primary microglia and in alveolar macrophages. (A) Representative confocal images of NOckout from LPS (1 μg/mL) primed J774A.1 macrophages in A647 (R) and DAR (G) channels in the absence (Upper) or presence (Lower) of NOS2 inhibitor 1400W. G/R intensities are represented as heat maps. (B) Representative confocal images of NOckoutmCpG from mouse primary microglia. Cells were incubated with NOckoutmCpG (500 nM) in the absence (Upper) or presence (Lower) of 1400W for 120 min in DMEM, imaged in DAR(G) and A647(R) channels, and converted into G/R heat maps. (C) Representative heat map (G/R) images of NOckout-, NOckoutiRNA-, and NOckoutRNA-treated J774A.1 cells. (D) Violin plot of the distribution of G/R values of ∼200 individual endosomes (n = 30 cells) of J774A.1 cells treated with VAS2870, ABAH, and 1400W in the presence and absence of LPS. (E) Violin plot of the distribution of G/R values of ∼100 individual endosomes (n = 20 cells) in primary microglia treated with NOckoutmCpG and NOckoutmGpC. (F) Violin plot of the distribution of G/R values of ∼200 individual endosomes (n = 30 cells) in J774A.1 macrophages treated with NOckoutRNA variants in the presence and absence of 1400W. All experiments were performed in triplicate. (Scale bar, 10 μm.) P values are obtained using Kruskal−Wallis statistical test across the dataset.

Article Snippet: Nitric oxide donor (DEANONOate), iNOS inhibitor (1400W), NADPH-oxidase inhibitor (VAS2870), MPO inhibitor (ABAH), and vacuolar H + -ATPase (V-ATPase) inhibitor (Bafilomycin A1) were purchased from Cayman Chemicals.

Techniques: Incubation

Journal: medRxiv

Article Title: Targeting myeloperoxidase to reduce neuroinflammation in X-linked dystonia parkinsonism

doi: 10.1101/2024.06.25.24309481

Figure Lengend Snippet: ( A ) Two-way ANOVA revealed a significant effect of treatment ([F(5, 36) = 20.08], p < 0.0001), genotype ([F(1, 36) = 2890], p<0.0001), and treatment x XDP interaction ([F(5, 36) = 22.71], p < 0.0001) on MPO activity in fibroblasts. Tukey’s’ test demonstrated a significant increase in MPO activity in XDP-compared to control-derived fibroblasts (p < 0.0001) as well as a significant increase in MPO activity in XDP-derived fibroblasts treated with either 0.1 (p < 0.0001), 0.5 (p < 0.0001), 1 (p < 0.0001), 5 (p < 0.0001), or 10 μg/mL of verdiperstat (p < 0.0001) compared to control-derived fibroblasts treated with the same doses of verdiperstat. Lastly, Tukey’s test revealed a significant decrease in MPO activity in XDP-derived fibroblasts treated with either 5 (p < 0.0001) or 10 μg/mL of verdiperstat (p < 0.0001) compared to vehicle-treated XDP-derived fibroblasts. (B) Two-way ANOVA revealed a significant effect of genotype ([F(4, 48) = 120.3], p < 0.001), and time ([F(12, 48) = 61.32], p < 0.0001) on ROS levels in fibroblasts. Tukey’s test revealed an expected significant increase in ROS in antimycin A treated fibroblasts compared to vehicle-treated control fibroblasts (p < 0.0001). There was also a significant increase in ROS in XDP fibroblasts compared to control fibroblasts (p < 0.0001). Importantly, Tukey’s test demonstrated a significant decrease in ROS in both control– and XDP-derived fibroblasts treated with verdiperstat compared to vehicle-treated control (p = 0.052) and XDP fibroblasts (p < 0.0001). (C) Two-way ANOVA demonstrated a significant effect of genotype ([F(4, 48) = 91.45], p < 0.0001), and time ([F(12, 48) = 3.579], p = 0.0008) on ROS levels in a second set of fibroblasts. As expected, there was a significant increase in antimycin A treated-fibroblasts compared to vehicle-treated control fibroblasts (p < 0.0001). Furthermore, Tukey’s test demonstrated a significant increase in ROS in XDP fibroblasts compared to control fibroblasts (p = 0.0025). Importantly, Tukey’s test revealed a significant decrease in ROS levels in both control– and XDP-derived fibroblasts treated with verdiperstat compared to vehicle-treated control (p < 0.0001) and XDP fibroblasts (p = 0.0002). ** p < 0.001; **** p<0.0001.

Article Snippet: Verdiperstat is a first-in-class, potent, selective, brain-permeable MPO inhibitor produced by Biohaven Pharmaceuticals, Inc.

Techniques: Activity Assay, Control, Derivative Assay

Journal: medRxiv

Article Title: Targeting myeloperoxidase to reduce neuroinflammation in X-linked dystonia parkinsonism

doi: 10.1101/2024.06.25.24309481

Figure Lengend Snippet: ( A ) One-way ANOVA revealed a significant effect of treatment ([F(2, 9) = 29.19], p = 0.0001) on MPO activity in SH-SY5Y cells. Tukey’s test revealed a significant increase in MPO activity in XDP treated cells compared to both vehicle (p = 0.0012) and XDP+verdiperstat treated cells (p=0.0001). (B) Two-way ANOVA revealed a significant effect of treatment ([F(4, 48) = 16.35], p < 0.0001), and time ([F(12, 48) = 31.68], p < 0.001) on ROS levels in SH-SY5Y cells. As expected, ROS levels were increased in antimycin A treated cells compared to vehicle treatment (Tukey’s test, p = 0.0003). Additionally, Tukey’s test demonstrated a significant increase in ROS in XDP treated cells compared to both vehicle (p = 0.0014) and XDP+verdiperstat treated cells (p < 0.0001, respectively). ** p<0.01; *** p<0.001; **** p<0.0001.

Article Snippet: Verdiperstat is a first-in-class, potent, selective, brain-permeable MPO inhibitor produced by Biohaven Pharmaceuticals, Inc.

Techniques: Activity Assay

Journal: Antioxidants

Article Title: Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity

doi: 10.3390/antiox10111791

Figure Lengend Snippet: Effect of MPO inhibition on NET formation. Neutrophils were stimulated with ( A ) PMA (20 nM), ( B ) P. aeruginosa PAO1 (MOI 10), ( C ) S. aureus (MOI 10), or ( D ) C. albicans (MOI 2) in the presence or absence of the MPO inhibitor thioxanthine 1 (TX1) (10 µM), and DNA fluorescence was measured after 4 h. The fluorescence of control (unstimulated) cells was subtracted from that of stimulated cells. Data are means (SE) of 4–8 separate experiments. *, significantly different than without TX1 ( p < 0.05 by paired t -test).

Article Snippet: The MPO inhibitors TX1 and AMZ198 were a gift from AstraZeneca (Gothenburg, Sweden).

Techniques: Inhibition, Fluorescence, Control

Journal: Antioxidants

Article Title: Neutrophil NET Formation with Microbial Stimuli Requires Late Stage NADPH Oxidase Activity

doi: 10.3390/antiox10111791

Figure Lengend Snippet: Early MPO activity is not sufficient to stimulate NET formation induced by PMA or P. aeruginosa PAO1. Neutrophils were stimulated with ( A ) PMA (20 nM) or ( B ) P. aeruginosa PAO1 (MOI 10). TX1 was added prior to or at the indicated times after stimulation. After 4 h, the presence of NETs was assessed by Sytox green plate assay. Data are presented as the percentage of fluorescent DNA of cells with stimulant alone (No TX1) and are means (SE) of 3–6 experiments with neutrophils from different donors. DNA fluorescence was significantly different than stimulated without TX1 ( p < 0.01 by one-way ANOVA followed by Dunnett’s multiple comparisons tests) at the following time points: PMA 0–30 min; P. aeruginosa PAO1 0–90 min. ( C , D ) Neutrophils were stimulated under the same conditions with the MPO inhibitor AZM-198. Data are means (SE) of three independent experiments with PMA and means with range of two separate experiments with P. aeruginosa PAO1. DNA fluorescence was significantly different than stimulated without AZM-198 ( p < 0.01 by one-way ANOVA followed by Dunnett’s multiple comparisons tests) only when AZM-198 was added prior to PMA.

Article Snippet: The MPO inhibitors TX1 and AMZ198 were a gift from AstraZeneca (Gothenburg, Sweden).

Techniques: Activity Assay, Fluorescence