Data_Sheet_1_Anion-Transport Mechanism of a Triazole-Bearing Derivative of Prodigiosine: A Candidate for Cystic Fibrosis Therapy.PDF

<p>Cystic fibrosis (CF) is a genetic lethal disease, originated from the defective function of the CFTR protein, a chloride and bicarbonate permeable transmembrane channel. CF mutations affect CFTR protein through a variety of molecular mechanisms which result in different functional defects. Current therapeutic approaches are targeted to specific groups of patients that share a common functional defect. We seek to develop an innovative therapeutic approach for the treatment of CF using anionophores, small molecules that facilitate the transmembrane transport of anions. We have characterized the anion transport mechanism of a synthetic molecule based on the structure of prodigiosine, a red pigment produced by bacteria. Anionophore-driven chloride efflux from large unilamellar vesicles is consistent with activity of an uniporter carrier that facilitates the transport of anions through lipid membranes down the electrochemical gradient. There are no evidences of transport coupling with protons. The selectivity sequence of the prodigiosin inspired EH160 ionophore is formate > acetate > nitrate > chloride > bicarbonate. Sulfate, phosphate, aspartate, isothionate, and gluconate are not significantly transported by these anionophores. Protonation at acidic pH is important for the transport capacity of the anionophore. This prodigiosin derived ionophore induces anion transport in living cells. Its low toxicity and capacity to transport chloride and bicarbonate, when applied at low concentration, constitute a promising starting point for the development of drug candidates for CF therapy.</p

Neuroprotective effect of MQ against Aβ_1–42 toxicity in rat neuronal cultures.

<p>Primary neuronal cultures of rat were simultaneously exposed to 1 µM Aβ_1–42 and MQ 0.5 or 1 µM after a 24 h sulforaphane (0.5 or 1 µM) pre-treatment. Whereas both the MQ concentrations partially protected from Aβ_1–42 toxicity following 0.5 µM sulforaphane exposure, the administration of 1 µM MQ after a pre-treatment with 1 µM sulforaphane was fully effective in protecting neuronal cultures from Aβ_1–42-induced neuronal death. Cell viability was assessed using the MTT (A) or LDH (B) assays. Data are expressed as mean ± S.E.M. Statistical analysis was performed using One-way ANOVA followed by Dunnett's multiple comparison test. *** p<0.001 compared with CTR; ^# p<0.05, ^## p<0.01, ^### p<0.001 compared with Aβ_1–42.</p

MQ rescues scopolamine-induced spatial memory deficits.

<p>Effect of MQ treatment on spontaneous alternation behavior (A) and MWM (B) in a scopolamine-induced amnesia model in mice. In the T-maze, MQ (7–15 mg/kg , p.o.) was administered 20 min before scopolamine (1 mg/kg, i.p.); in the MWM, MQ (7 and 15 mg/kg, p.o.) was administered 20 min before scopolamine (1.5 mg/kg, i.p.). Data are expressed as mean ± S.E.M. Statistical analysis was performed using One-way ANOVA followed by post-hoc Tukey's test where appropriate, *** p<0.001 vs. controls (Veh); ^# p<0.05, ^## p<0.01 vs. scopolamine-treated animals (Scop).</p

Beneficial effect of MQ on Aβ_1–42 i.c.v. treated animals.

<p>Effect of MQ daily treatments in the spontaneous alternation (A) and passive avoidance task (B) using a Aβ- induced amnesia model in mice. 800 pmol Aβ_1–42 and Aβ_42-1 or their vehicle (PBS) were i.c.v. injected 7 and 8 days before T-maze and passive avoidance, respectively. MQ 15 mg/kg p.o. was administered daily for 2, 4, and 6 days before behavioral testing. Data are expressed as mean ± S.E.M. Statistical analysis was performed using One-way ANOVA followed by post-hoc Tukey's test. * p<0.05, *** p<0.001 vs. controls (PBS and Aβ_42-1); ^# p<0.05, ^## p<0.001 vs. amyloid-injected animals.</p

A Potent Systemically Active <i>N</i>‑Acylethanolamine Acid Amidase Inhibitor that Suppresses Inflammation and Human Macrophage Activation

Fatty acid ethanolamides such as palmitoylethanolamide (PEA) and oleoylethanolamide (OEA) are lipid-derived mediators that potently inhibit pain and inflammation by ligating type-α peroxisome proliferator-activated receptors (PPAR-α). These bioactive substances are preferentially degraded by the cysteine hydrolase, <i>N</i>-acylethanolamine acid amidase (NAAA), which is highly expressed in macrophages. Here, we describe a new class of β-lactam derivatives that are potent, selective, and systemically active inhibitors of intracellular NAAA activity. The prototype of this class deactivates NAAA by covalently binding the enzyme’s catalytic cysteine and exerts profound anti-inflammatory effects in both mouse models and human macrophages. This agent may be used to probe the functions of NAAA in health and disease and as a starting point to discover better anti-inflammatory drugs

Hit Optimization of 5‑Substituted‑<i>N</i>‑(piperidin-4-ylmethyl)‑1<i>H</i>‑indazole-3-carboxamides: Potent Glycogen Synthase Kinase‑3 (GSK-3) Inhibitors with in Vivo Activity in Model of Mood Disorders

Novel treatments for bipolar disorder with improved efficacy and broader spectrum of activity are urgently needed. Glycogen synthase kinase 3β (GSK-3β) has been suggested to be a key player in the pathophysiology of bipolar disorder. A series of novel GSK-3β inhibitors having the common <i>N</i>-[(1-alkylpiperidin-4-yl)­methyl]-1<i>H</i>-indazole-3-carboxamide scaffold were prepared taking advantage of an X-ray cocrystal structure of compound <b>5</b> with GSK-3β. We probed different substitutions at the indazole 5-position and at the piperidine-nitrogen to obtain potent ATP-competitive GSK-3β inhibitors with good cell activity. Among the compounds assessed in the <i>in vivo</i> PK experiments, <b>14i</b> showed, after i.p. dosing, encouraging plasma PK profile and brain exposure, as well as efficacy in a mouse model of mania. Compound <b>14i</b> was selected for further <i>in vitro</i>/<i>in vivo</i> pharmacological evaluation, in order to elucidate the use of ATP-competitive GSK-3β inhibitors as new tools in the development of new treatments for mood disorders