Journal: Clinical Microbiology Reviews

Article Title: Vaccine and Therapeutic Options To Control Chikungunya Virus

doi: 10.1128/CMR.00104-16

Figure Lengend Snippet: Therapeutic candidates under development and primary characteristics of each candidate a

Article Snippet: Host cell targets , Prostratin , BGM, Vero, HEL, ATCC CRL-2522 cell culture , CHIKV-899 (ECSA) , NP , 0.2 to 8 μM , >100 μM , Decreased viral RNA levels , NP , ND , ND , 123.

Techniques: Functional Assay, Concentration Assay, Inhibition, In Vivo, Cell Culture, Plasmid Preparation, Binding Assay, Infection, In Vitro

Journal: Nature Communications

Article Title: Drug repurposing screens identify chemical entities for the development of COVID-19 interventions

doi: 10.1038/s41467-021-23328-0

Figure Lengend Snippet: a Simplified assay workflow. b Representative images from dimethyl sulfoxide (DMSO)-, remdesivir- or apilimod-treated wells. The entire imaged area per well (four fields of view taken with a ×10 objective and stitched together) is shown for each treatment, as well as an 8-fold magnified segment demarcated with a white box. DNA signal [4′,6-diamidino-2-phenylindole (DAPI)] is colored green, and the virus visualized with immunofluorescence is colored magenta. Infected (arrow) and uninfected (arrowhead) cells are indicated; 500 µm and 50 µm scale bars are shown in the composite and magnified images, respectively. Raw and normalized (Norm.) values calculated from the images are shown. c Box and whiskers plot of SARS-CoV-2 assay control EC 50 s obtained from n = 9 independent biological experiments with all data points shown. Whiskers indicate minimums and maximums, the box extends from the 25th to 75th percentile, the center of the box is the median and geomeans are reported. d Heat map images of normalized data from 1.9 µM ReFRAME screening plates. Normalized activity values for percentage infected cells and total cell numbers are indicated according to the scale bar and density plot for compound and control wells is shown. DMSO-treated wells are in column 24 and positive control-treated wells (blocks of wells with 2.4 µM remdesivir, 2.4 µM apilimod, or 9.6 µM puromycin) in column 23. Density plots representing the frequency of values associated with each well type are shown on the right. e Distribution of 1.9 µM ReFRAME screen data for compound and control wells. f Screen hit selection thresholds. g Library dose-response reconfirmation results, with the SARS-CoV-2 EC 50 of each compound plotted against its host-cell toxicity CC 50 as assessed in uninfected HeLa-ACE2 cells. Gray area defines non-selective compounds, where the selectivity index (CC 50 /EC 50 ) is < 10. Dotted lines represent maximal concentrations tested in dose-response studies for the assay compounds (40 µM) and controls apilimod and remdesivir (9.6 µM). Activities of controls (black diamonds) and assay compounds (pink diamonds) are shown. Activity of the ReFRAME library copy of puromycin that was screened as part of this hit reconfirmation is also indicated (red diamond). Source data are provided as a Source Data file.

Article Snippet: HeLa-ACE2 data were normalized to neutral (DMSO) minus inhibitor controls (2.4 µM remdesivir for antiviral effect in HeLa-ACE2 cells and 9.6 µM puromycin for infected host-cell toxicity).

Techniques: Immunofluorescence, Infection, Activity Assay, Positive Control, Selection

Journal: Nature Communications

Article Title: Drug repurposing screens identify chemical entities for the development of COVID-19 interventions

doi: 10.1038/s41467-021-23328-0

Figure Lengend Snippet: a Heat map of activities of HeLa-ACE2 ReFRAME hits reconfirmed with fresh powder stocks in HeLa-ACE2 and Calu-3 cells (EC 50 , CC 50 , selectivity index (SI)) and the time of addition (TOA) vs standard 24 h assay HeLa-ACE2 antiviral EC 50 ratio. Double asterisk indicates two compounds identified in primary screening that were not selective in powder reconfirmation in HeLa-ACE2 cells but were active and selective in Calu-3 cells. X indicates missing data. b Heat map of activities of Calu-3 ReFRAME hits reconfirmed with fresh powder stocks in Calu-3 and HeLa-ACE2 cells (EC 50 , CC 50 , SI). Double asterisk indicates compounds with antiviral efficacy in Calu-3 < 80%. All raw data from experiments graphically represented in a and b are reported as part of Supplementary Data and and include the independent number each measurement was performed (≥ 3 for most compounds/assays). c HeLa-ACE2 TOA time course showing SARS-CoV-2 % infected cells at different times post-infection. Cells were infected with virus for one hour, washed extensively prior to compound treatment, and fixed at the indicates times. The 10 hpi timepoint was chosen for the TOA assay. Means ± s.d. are shown, n = 3 technical replicates examined over one independent experiment. d The composition of the ReFRAME repurposing library with respect to clinical stage of development and disease indication. e Classification of HeLa-ACE2 and Calu-3 potent and selective hits according to their functional annotation. Source data are provided as a Source Data file.

Article Snippet: HeLa-ACE2 data were normalized to neutral (DMSO) minus inhibitor controls (2.4 µM remdesivir for antiviral effect in HeLa-ACE2 cells and 9.6 µM puromycin for infected host-cell toxicity).

Techniques: Infection, Functional Assay

Journal: Nature Communications

Article Title: Drug repurposing screens identify chemical entities for the development of COVID-19 interventions

doi: 10.1038/s41467-021-23328-0

Figure Lengend Snippet: a Representative dose-response curves of assay controls ( a ) and MK-4482 prodrug and its parent, N-hydroxycytidine ( b ) from three or two independent experiments in HeLa-ACE2 and Calu-3 assays. Individual values of technical triplicates, compound structures, and geomeans for each infection assay (SARS-CoV-2 EC 50 and total cell count EC 50 ) and uninfected cytotoxicity CC 50 are shown. HeLa-ACE2 TOA EC 50 geomeans are also shown as applicable. EC 50 s or CC 50 s < 200 nM are marked with dark green background, < 1000 nM with light green, and < 9000 nM with yellow. Inactive compounds or with EC 50 s or CC 50 s > 9000 nM are marked with pink. ↑ indicates an EC 50 for an increasing dose-response curve slope. c Viral log reduction in apical supernatants determined with RT-qPCR and percentage toxicity determined with an LDH assay in ALI HBEC SARS-CoV-2 infection model following a 3-day treatment with 5 µM of each indicated compound. Dark green cell background indicates a > 2-fold log reduction compared to neutral controls, light green indicates a > 1-fold log reduction, yellow a < 1-fold log reduction, and pink indicates a viral load increase. d Diagram illustrating differences between compound activities observed in different cell infection models as related to different entry mechanisms available in each cell line. Source data are provided as a Source Data file.

Article Snippet: HeLa-ACE2 data were normalized to neutral (DMSO) minus inhibitor controls (2.4 µM remdesivir for antiviral effect in HeLa-ACE2 cells and 9.6 µM puromycin for infected host-cell toxicity).

Techniques: Infection, Cell Counting, Quantitative RT-PCR, Lactate Dehydrogenase Assay

Journal: Nature Communications

Article Title: Drug repurposing screens identify chemical entities for the development of COVID-19 interventions

doi: 10.1038/s41467-021-23328-0

Figure Lengend Snippet: Attractive reconfirmed hits with activity and selectivity against SARS-CoV-2.

Article Snippet: HeLa-ACE2 data were normalized to neutral (DMSO) minus inhibitor controls (2.4 µM remdesivir for antiviral effect in HeLa-ACE2 cells and 9.6 µM puromycin for infected host-cell toxicity).

Techniques: Activity Assay, Protease Inhibitor

Journal: Molecules

Article Title: Synthesis and Pharmacological Effects of Diosgenin–Betulinic Acid Conjugates

doi: 10.3390/molecules25153546

Figure Lengend Snippet: Cytotoxicity screening tests (IC 50 values (μM), 72 h).

Article Snippet: The compounds 1 – 8 were subjected to the cytotoxicity screening tests on cells of human T-lymphoblastic leukemia (CEM), cells of human breast adenocarcinoma (MCF7), cells of human cervical cancer (HeLa), and human colon carcinoma (HCT 116 cancer cell lines), using normal human fibroblasts (BJ) as reference cell lines.

Techniques: