Genetic or pharmacological ablation of MAGL reduces fever response in peripheral LPS-induced fever model.

<p><b>(A)</b> CBT profile following i.p. injection of LPS (100 μg/kg) or vehicle (Saline) of <i>Mgll</i>^<i>+/+</i> mice treated i.p. with JZL184 (40 mg/kg). *p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh vs. <i>Mgll</i>^<i>+/+</i> + LPS; ^#p<0.05, <i>Mgll</i>^<i>+/+</i>+ LPS vs. <i>Mgll</i>^<i>-/-</i> + LPS; ^&p<0.05, <i>Mgll</i>^<i>+/+</i> + Veh vs. <i>Mgll</i>^<i>-/-</i> + LPS. <b>(B)</b> CBT profile of <i>Mgll</i>^<i>-/-</i> and <i>Mgll</i>^<i>+/+</i> mice following i.p. injection of LPS (100 μg/kg) or vehicle (Saline) as indicated. Injection was performed at time 0. Data are shown as mean ± sem, n = 6 mice per group, *p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh + Veh vs. <i>Mgll</i>^<i>+/+</i> + Veh + LPS; # p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh+ LPS vs. <i>Mgll</i>^<i>+/+</i>+ JZL184 + LPS.</p

Anti-pyrogenic effects of MAGL inhibitors are independent of CB1 cannabinoid receptor activity.

<p>CBT profile following i.p. injection of LPS (100 μg/kg) of <i>Mgll</i>^<i>+/+</i> mice receiving i.p. injection of rimonabant (1 mg/kg) and/ or JZL184 (40 mg/kg). Rimonabant did not affect the hypothermic effects of JZL184. Data are shown as mean ± sem, n = 6 mice per group, *p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh + Veh + Veh vs. <i>Mgll</i>^<i>+/+</i>+ Veh + Veh + LPS; ^#p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh + Veh + LPS vs. <i>Mgll</i>^<i>+/+</i>+ JZL184 + Veh + LPS; ^&p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh + Rim + LPS vs. <i>Mgll</i>^<i>+/+</i>+ Veh + Rim + Veh; ^$p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh + Rim + LPS vs. <i>Mgll</i>^<i>+/+</i>+ JZL184 + Rim + LPS. p>0.05 (NS) <i>Mgll</i>^<i>+/+</i>+ Veh + Veh + Veh vs <i>Mgll</i>^<i>+/+</i>+ Veh + Rim + Veh, <i>Mgll</i>^<i>+/+</i>+ JZL184 + Rim + LPS vs <i>Mgll</i>^<i>+/</i> + JZL184 + Veh + LPS, <i>Mgll</i>^<i>+/+</i>+ Veh + Veh + LPS vs <i>Mgll</i>^<i>+/+</i>+ Veh + Rim + LPS.</p

Genetic or pharmacological ablation of MAGL reduces fever response in central IL-1β-induced fever model.

<p><b>(A)</b> CBT profile following icv injection into POA of IL-1β (500 pg/0.5 μl) or aCSF of <i>Mgll</i>^<i>+/+</i> mice treated i.p. with JZL184 (40 mg/kg). *p<0.05, <i>Mgll</i>^<i>+/+</i> + Veh vs. <i>Mgll</i>^<i>+/+</i> + IL-1β; ^#p<0.05 <i>Mgll</i>^<i>+/</i> + IL-1β vs. <i>Mgll</i>^<i>-/-</i>+ IL-1β. <b>(B)</b> CBT profile of <i>Mgll</i>^<i>-/-</i> and <i>Mgll</i>^<i>+/+</i> mice following icv injection into POA of IL-1β (500 pg/0.5 μl) or aCSF as indicated. Injection was performed at time 0. Data are shown as mean ± sem, n = 6 mice per group. *p<0.05, <i>Mgll</i>^<i>+/+</i>+ Veh + Veh vs. <i>Mgll</i>^<i>+/+</i>+ Veh + IL-1β; ^#p<0.05 <i>Mgll</i>^<i>+/+</i>+ Veh+ IL-1β vs. <i>Mgll</i>^<i>+/+</i>+ JZL184 + IL-1β.</p

<i>Mgll</i>^<i>-/-</i> and <i>Mgll</i>^<i>+/+</i> mice have similar core body temperature profiles.

<p>CBT profile of <i>Mgll</i>^<i>-/-</i> and <i>Mgll</i>^<i>+/+</i> male mice over 24 hrs. No statistically significant differences were observed across genotypes. Data are shown as mean ± sem, n = 6 mice per group, p>0.05.</p

Development of novel NEMO-binding domain mimetics for inhibiting IKK/NF-κB activation

<div><p>Nuclear factor κB (NF-κB) is a transcription factor important for regulating innate and adaptive immunity, cellular proliferation, apoptosis, and senescence. Dysregulation of NF-κB and its upstream regulator IκB kinase (IKK) contributes to the pathogenesis of multiple inflammatory and degenerative diseases as well as cancer. An 11–amino acid peptide containing the NF-κB essential modulator (NEMO)-binding domain (NBD) derived from the C-terminus of β subunit of IKK, functions as a highly selective inhibitor of the IKK complex by disrupting the association of IKKβ and the IKKγ subunit NEMO. A structure-based pharmacophore model was developed to identify NBD mimetics by in silico screening. Two optimized lead NBD mimetics, SR12343 and SR12460, inhibited tumor necrosis factor α (TNF-α)- and lipopolysaccharide (LPS)-induced NF-κB activation by blocking the interaction between IKKβ and NEMO and suppressed LPS-induced acute pulmonary inflammation in mice. Chronic treatment of a mouse model of Duchenne muscular dystrophy (DMD) with SR12343 and SR12460 attenuated inflammatory infiltration, necrosis and muscle degeneration, demonstrating that these small-molecule NBD mimetics are potential therapeutics for inflammatory and degenerative diseases.</p></div

The IC₅₀ of NBD mimetics.

<p>Inhibitory effects of NBD mimetics on NF-κB activation were measured by luciferase assays at multiple concentrations: 0, 25, 50, 100, and 150 μM. IC₅₀ of NBD mimetics was determined based on the dose-dependent curve by using GraphPad.</p

Optimization of 2‑Anilino 4‑Amino Substituted Quinazolines into Potent Antimalarial Agents with Oral in Vivo Activity

Novel antimalarial therapeutics that target multiple stages of the parasite lifecycle are urgently required to tackle the emerging problem of resistance with current drugs. Here, we describe the optimization of the 2-anilino quinazoline class as antimalarial agents. The class, identified from publicly available antimalarial screening data, was optimized to generate lead compounds that possess potent antimalarial activity against <i>P. falciparum</i> parasites comparable to the known antimalarials, chloroquine and mefloquine. During the optimization process, we defined the functionality necessary for activity and improved <i>in vitro</i> metabolism and solubility. The resultant lead compounds possess potent activity against a multidrug resistant strain of <i>P. falciparum</i> and arrest parasites at the ring phase of the asexual stage and also gametocytogensis. Finally, we show that the lead compounds are orally efficacious in a 4 day murine model of malaria disease burden

Hematological measures in acutely treated mice.

<p>Hematological measures in acutely treated mice.</p

Optimization of 2‑Anilino 4‑Amino Substituted Quinazolines into Potent Antimalarial Agents with Oral in Vivo Activity

Novel antimalarial therapeutics that target multiple stages of the parasite lifecycle are urgently required to tackle the emerging problem of resistance with current drugs. Here, we describe the optimization of the 2-anilino quinazoline class as antimalarial agents. The class, identified from publicly available antimalarial screening data, was optimized to generate lead compounds that possess potent antimalarial activity against <i>P. falciparum</i> parasites comparable to the known antimalarials, chloroquine and mefloquine. During the optimization process, we defined the functionality necessary for activity and improved <i>in vitro</i> metabolism and solubility. The resultant lead compounds possess potent activity against a multidrug resistant strain of <i>P. falciparum</i> and arrest parasites at the ring phase of the asexual stage and also gametocytogensis. Finally, we show that the lead compounds are orally efficacious in a 4 day murine model of malaria disease burden

NBD mimetics improve muscular pathology and grip strength in mdx mice.

<p>(A) Shown is the treatment regimen with NBD mimetics in mdx mice. (B) Hematoxylin–eosin staining of TA muscle from 7-wk-old, treated or untreated mdx mice. Images were taken at magnification of 20×. Representative images were shown for each treatment group. (C) Quantitation of the percentage of area exhibiting necrosis or infiltration by using Image J. (D) qRT-PCR analysis of eMyHC and Pax7 in TA muscle. (E) TA muscle from 7-wk-old, treated or untreated mdx mice was stained with laminin by IHC to outline myofibers. Overall images of TA muscle were shown on the left side, and the inset panel images were shown on the right side. (F) Quantitation of minimum Feret diameter in centrally and noncentrally nucleated myofibers. (G) Forelimb force determined by grip strength test. Five-wk-old (2 wk post treatment) and 7-wk-old (4 wk post treatment) mdx mice, treated or untreated, were tested for forelimb grip strength. Five sequential tests were performed, and the force was normalized to body weight. Data were shown as mean +/− SEM. Dosages used are as follows: SR12343 (30 mg/kg), SR12460 (30 mg/kg), and 8K-NBD peptide (10 mg/kg). <i>n</i> = 6–7 each group. *<i>P</i> < 0.05, **<i>P</i> < 0.01. Underlying data can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.2004663#pbio.2004663.s001" target="_blank">S1 Data</a>. eMyHC, embryonic myosin heavy chain; IHC, immunohistochemistry; NBD, NEMO-binding domain; NEMO, NF-κB essential modulator; Pax7, paired box protein 7; qRT-PCR, quantitative real-time polymerase chain reaction; TA, tibialis anterior.</p