Screening a small molecule library to identify inhibitors of NF-κB inducing kinase and pro-labor genes in human placenta.

The non-canonical NF-κB signaling (RelB/p52) pathway drives pro-labor genes in the human placenta, including corticotropin-releasing hormone (CRH) and cyclooxygenase-2 (COX-2), making this a potential therapeutic target to delay onset of labor. Here we sought to identify small molecule compounds from a pre-existing chemical library of orally active drugs that can inhibit this NF-κB signaling, and in turn, human placental CRH and COX-2 production. We used a cell-based assay coupled with a dual-luciferase reporter system to perform an in vitro screening of a small molecule library of 1,120 compounds for inhibition of the non-canonical NF-κB pathway. Cell toxicity studies and drug efflux transport MRP1 assays were used to further characterize the lead compounds. We have found that 14 drugs have selective inhibitory activity against lymphotoxin beta complex-induced activation of RelB/p52 in HEK293T cells, several of which also inhibited expression of CRH and COX-2 in human term trophoblast. We identified sulfapyridine and propranolol with activity against CRH and COX-2 that deserve further study. These drugs could serve as the basis for development of orally active drugs to affect length of gestation, first in an animal model, and then in clinical trials to prevent preterm birth during human pregnancy.

Novel strategy for sulfapyridine detection using a fully integrated electrochemical Bio-MEMS: Application to honey analysis.

Sulfapyridine (SPy) is a sulfonamide antibiotic largely employed as veterinary drugs for prophylactic and therapeutic purposes. Therefore, its spread in the food products has to be restricted. Herein, we report the synthesis and characterization of a novel electrochemical biosensor based on gold microelectrodes modified with a new structure of magnetic nanoparticles (MNPs) coated with poly(pyrrole-co-pyrrole-2-carboxylic acid) (Py/Py-COOH) for high efficient detection of SPy. This analyte was quantified through a competitive detection procedure with 5-[4-(amino)phenylsulfonamide]-5-oxopentanoic acid-BSA (SA2-BSA) antigens toward polyclonal antibody (Ab-155). Initially, gold working electrodes (WEs) of integrated biomicro electro-mechanical system (BioMEMS) were functionalized by Ppy-COOH/MNPs, using a chronoamperometric (CA) electrodeposition. Afterward, SA2-BSA was covalently bonded to Py/Py-COOH/MNP modified gold WEs through amide bonding. The competitive detection of the analyte was made by a mixture of a fixed concentration of Ab-155 and decreasing concentrations of SPy from 50µgL-1 to 2ngL-1. Atomic Force Microscopy characterization was performed in order to ensure Ppy-COOH/MNPs electrodeposition on the microelectrode surfaces. Electrochemical measurements of SPy detection were carried out using electrochemical impedance spectroscopy (EIS). This biosensor was found to be highly sensitive and specific for SPy, with a limit of detection of 0.4ngL-1. This technique was exploited to detect SPy in honey samples by using the standard addition method. The measurements were highly reproducible for detection and interferences namely, sulfadiazine (SDz), sulfathiazole (STz) and sulfamerazine (SMz). Taking these advantages of sensitivity, specificity, and low cost, our system provides a new horizon for development of advanced immunoassays in industrial food control.