Reversible and irreversible inhibitors of coronavirus Nsp15 endoribonuclease.

The emergence of severe acute respiratory syndrome coronavirus 2, the causative agent of coronavirus disease 2019, has resulted in the largest pandemic in recent history. Current therapeutic strategies to mitigate this disease have focused on the development of vaccines and on drugs that inhibit the viral 3CL protease or RNA-dependent RNA polymerase enzymes. A less-explored and potentially complementary drug target is Nsp15, a uracil-specific RNA endonuclease that shields coronaviruses and other nidoviruses from mammalian innate immune defenses. Here, we perform a high-throughput screen of over 100,000 small molecules to identify Nsp15 inhibitors. We characterize the potency, mechanism, selectivity, and predicted binding mode of five lead compounds. We show that one of these, IPA-3, is an irreversible inhibitor that might act via covalent modification of Cys residues within Nsp15. Moreover, we demonstrate that three of these inhibitors (hexachlorophene, IPA-3, and CID5675221) block severe acute respiratory syndrome coronavirus 2 replication in cells at subtoxic doses. This study provides a pipeline for the identification of Nsp15 inhibitors and pinpoints lead compounds for further development against coronavirus disease 2019 and related coronavirus infections.

HPLC Method for Simultaneous Determination of Bisphenol-A-Diglycidyl Ether and Some of Its Reaction Products in Canned Foods Using Photodiode Array Detector.

Gradient reversed-phase high-performance liquid chromatography with photodiode array detection was used for separation, detection and quantification of bisphenol-A-diglycidyl ether (BADGE) and some of its reaction products, namely, BADGE·HCl·H2O, BADGE·H2O and BADGE·2HCl in pure form and in canned foods, where canned beans and tuna were used as representatives of aqueous and oil-in-water food matrices, respectively. The proposed method had a linear range of 0.01-0.5 µg g-1 for BADGE·HCl·H2O, BADGE·H2O, BADGE·2HCl and 0.02-0.7 µg g-1 for BADGE in aqueous food matrices. In oil-in-water matrices, the method was proven to be sensitive over a linear range of 0.01-0.5 µg g-1 for BADGE·HCl·H2O, BADGE·H2O and 0.02-0.7 µg g-1 for BADGE·2HCl and BADGE. The limits of detection and quantification ranged from 0.24 to 1.22 ng g-1 and 0.73 to 14.07 ng g-1, respectively. Excellent intraday and interday precision (n = 9) were obtained with RSD% of 0.84-2.19% and 1.88-2.52%, respectively. Accuracy was measured at five concentration levels and the recoveries ranged from 96.31% to 98.76% with an acceptable variation of ±0.9-2.87. Results suggest that the proposed method could be applied for the routine analysis of the studied compounds in their laboratory-prepared mixtures and in various types of canned foods following the limits and regulations of the European Union.

Validated chemometrics-assisted spectrophotometric methods for simultaneous determination of bisphenol-A-diglycidyl ether and some of its reaction products in canned foods in the Egyptian market.

Two multivariate calibration methods, namely principal component regression (PCR) and partial least squares (PLS-2) have been developed, validated and compared for the simultaneous determination of bisphenol-A-diglycidyl ether (BADGE) and some of its reaction products, including BADGE·HCl·H2O, BADGE·H2O and BADGE·2HCl. Chemometrics allowed rapid, accurate and precise simultaneous quantification of the analytes of interest which was not possible by other spectrophotometric methods due to their severe spectral overlap. PCR and PLS-2 techniques successfully quantified BADGE, BADGE·HCl·H2O, BADGE·H2O and BADGE·2HCl in the ranges of 1.4-3.4, 1-5, 1-4.2 and 1-7 µg mL-1, respectively. The constructed models were validated according to the International Conference on Harmonization guidelines and successfully applied for the determination of these compounds in pure form, laboratory prepared mixtures and in various types of canned foods following the limits and regulations of the European Union (EU) where satisfactory recovery results were obtained.