Serendipitous discovery of 2-((phenylsulfonyl)methyl)-thieno[3,2-d]pyrimidine derivatives as novel HIV-1 replication inhibitors.

We identified a novel class of 2-((phenylsulfonyl)methyl)-thieno[3,2-d]pyrimidine compounds as potent HIV-1 replication inhibitors serendipitously during the process of evaluation of triazolothienopyrimidine (TTPM) compounds. Herein, we report synthesis and biological evaluation of 2-((phenylsulfonyl)methyl)-thieno[3,2-d]pyrimidine compounds using a cell-based full replication assay to identify thienopyrimidines 6 and 30, which could be further utilized as viable lead compounds.

Identification of a novel sulfonamide non-nucleoside reverse transcriptase inhibitor by a phenotypic HIV-1 full replication assay.

Classical target-based, high-throughput screening has been useful for the identification of inhibitors for known molecular mechanisms involved in the HIV life cycle. In this study, the development of a cell-based assay that uses a phenotypic drug discovery approach based on automated high-content screening is described. Using this screening approach, the antiviral activity of 26,500 small molecules from a relevant chemical scaffold library was evaluated. Among the selected hits, one sulfonamide compound showed strong anti-HIV activity against wild-type and clinically relevant multidrug resistant HIV strains. The biochemical inhibition, point resistance mutations and the activity of structural analogs allowed us to understand the mode of action and propose a binding model for this compound with HIV-1 reverse transcriptase.

Synthesis and biological evaluation of triazolothienopyrimidine derivatives as novel HIV-1 replication inhibitors.

We identified a novel class of triazolothienopyrimidine (TTPM) compounds as potent HIV-1 replication inhibitors during a high-throughput screening campaign that evaluated more than 200,000 compounds using a cell-based full replication assay. Herein, we report the optimization of the antiviral activity in a cell-based assay system leading to the discovery of aryl-substituted TTPM derivatives (38, 44, and 45), which exhibited significant inhibition of HIV-1 replication with acceptable safety margins. These novel and potent TTPMs could serve as leads for further development.

A novel 3,4-dihydropyrimidin-2(1H)-one: HIV-1 replication inhibitors with improved metabolic stability.

Following the previous SAR of a novel dihydropyrimidinone scaffold as HIV-1 replication inhibitors a detailed study directed towards optimizing the metabolic stability of the ester functional group in the dihydropyrimidinone (DHPM) scaffold is described. Replacement of the ester moiety by thiazole ring significantly improved the metabolic stability while retaining antiviral activity against HIV-1 replication. These novel and potent DHPMs with bioisosteres could serve as advanced leads for further optimization.

Discovery of 3,4-dihydropyrimidin-2(1H)-ones with inhibitory activity against HIV-1 replication.

3,4-Dihydropyrimidin-2(1H)-ones (DHPMs) were selected and derivatized through a HIV-1 replication assay based on GFP reporter cells. Compounds 14, 25, 31, and 36 exhibited significant inhibition of HIV-1 replication with a good safety profile. Chiral separation of each enantiomer by fractional crystallization showed that only the S enantiomer retained anti-HIV activity. Compound (S)-40, a novel and potent DHPM analog, could serve as an advanced lead for further development and the determination of the mechanism of action.

Sterols from a soft coral, Dendronephthya gigantea as farnesoid X-activated receptor antagonists.

Three new steroids 3-oxocholest-1,22-dien-12ß-ol (1), 3-oxocholest-1,4-dien-20ß-ol (2), 3-oxocholest-1,4-dien-12ß-ol (3), and three known steroids (20S)-20-Hydroxyergosta-1,4,24(28)-trien-3-one (4) [7a], 5α,8α-Epidioxycholesta-6,22-dien-3ß-ol (5) [10] and 5-cholestene-3ß,12ß-diol (6) [11] were isolated from a soft coral Dendronephthya gigantea. Their chemical structures and relative stereochemistry were elucidated by the analysis of HRMS and 2-D NMR spectroscopic data. The steroids 1 and 2 showed notable inhibitory activity against farnesoid X-activated receptor (FXR) with IC(50)'s 14 and 15µM.

Discovery of Phenylaminopyridine Derivatives as Novel HIV-1 Non-nucleoside Reverse Transcriptase Inhibitors.

We identified a novel class of aryl-substituted triazine compounds as potent non-nucleoside reverse transcriptase inhibitors (NNRTIs) during a high-throughput screening campaign that evaluated more than 200000 compounds for antihuman immunodeficiency virus (HIV) activity using a cell-based full replication assay. Herein, we disclose the optimization of the antiviral activity in a cell-based assay system leading to the discovery of compound 27, which possessed excellent potency against wild-type HIV-1 (EC50 = 0.2 nM) as well as viruses bearing Y181C and K103N resistance mutations in the reverse transcriptase gene. The X-ray crystal structure of compound 27 complexed with wild-type reverse transcriptase confirmed the mode of action of this novel class of NNRTIs. Introduction of a chloro functional group in the pyrazole moiety dramatically improved hERG and CYP inhibition profiles, yielding highly promising leads for further development.

Quantitative detection of HIV-1 particles using HIV-1 neutralizing antibody-conjugated beads.

The human immunodeficiency virus (HIV) pandemic mainly affects developing countries, where the Joint United Nations Programme on HIV/AIDS (UNAIDS) estimates suggest that less than 1 in 10 people are aware of their HIV sero-status. In order to enhance epidemiological surveys, prevention programs, and therapeutic interventions, development of specific, rapid, and convenient diagnostic detection systems is still warranted. Here we report the direct detection of HIV particles using broadly HIV-1 neutralizing gp120 monoclonal antibody (gp120MAbs)-conjugated magnetic beads (MBs) and fluorescent nanosized polymeric beads (FNBs). The HIV-1 envelope glycoprotein gp120 is anchored to the viral surface through gp41 and mediates entry into target cells by interaction with the main cellular receptor (CD4) and coreceptors (e.g., CCR5 and CXCR4). FNBs conjugated to gp120MAbs (gp120MAbs-FNBs) were used to generate fluorescent signals, whereas MBs conjugated to gp120MAbs (gp120MAbs-MBs) were employed to isolate HIV-1 particles. In presence of HIV-1 particles, addition of gp120MAbs-FNBs and gp120MAbs-MBs leads to the formation of a MBs/HIV-1 particles/FNB complex, which can be easily isolated and concentrated by common magnet separation. We demonstrate the ability of detecting HIV-1 particles specifically and directly using MBs and FNBs with low sample volume (less than 100 microL) and rapidity (less than 1.5 h) without any pretreatment of test samples. The specific binding of FNBs with HIV-1 particles on the surface of MBs was confirmed by fluorescence microscopy and fluorescence-activated cell sorting (FACS). Imaging and FACS analysis revealed the specific and quantitative detection of HIV-1 particles. These results provide proof-of-principle that broadly HIV-1 neutralizing gp120 antibodies coupled to nanobeads can be employed for the direct detection of HIV-1 particles with potential implication for the development of specific, rapid, and convenient diagnostic systems.

Scalarane sesterterpenes from a marine sponge of the genus Spongia and their FXR antagonistic activity.

Three new scalarane-based sesterterpenes, 1- 3, were isolated from a marine sponge of the genus Spongia, and their chemical structures were elucidated by analysis of HRMS and 2-D NMR spectra. The isolated compounds 1 and 3 showed inhibition against the farnesoid X-activated receptor (FXR) with IC50 values of 2.4 and 24 microM, respectively. In particular, compound 3 directly inhibited the interaction between FXR and a coactivator peptide (SRC-1) as determined by surface plasmon resonance (SPR) spectroscopy.