Metformin, dimethyl sulfoxide (DMSO), glycerol, glycine, sodium chloride, sodium dodecyl sulfate, Trizma base, and Tween20 were purchased from Sigma (St. Louis, MO, USA). The FDA-approved drug library (SCREEN-WELL® FDA-approved drug library V2) was purchased from Enzo Life Sciences (Farmingdale, NY, USA).
Screen well fda approved drug library v2
Manufactured by Enzo Life Sciences
Sourced in United States
The Screen-Well® FDA Approved Drug Library V2 is a collection of 1,581 compounds that have been approved by the U.S. Food and Drug Administration (FDA) for human use. This library is designed to facilitate drug discovery and repurposing studies.
Automatically generated - may contain errors
Lab products found in correlation
Trizma base, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Glycerol, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Metformin, by Merck Group (1 mentions)
Glycine, by Merck Group (1 mentions)
In cell elisa colorimetric detection kit, by Thermo Fisher Scientific (1 mentions)
C2c12 myoblast, by American Type Culture Collection (1 mentions)
Hrp conjugated igg secondary antibody, by Jackson ImmunoResearch (1 mentions)
Oligonucleotide, by Integrated DNA Technologies (1 mentions)
Dasatinib, by Santa Cruz Biotechnology (1 mentions)
Nilotinib, by Santa Cruz Biotechnology (1 mentions)
Bosutinib, by Santa Cruz Biotechnology (1 mentions)
Imatinib mesylate, by Santa Cruz Biotechnology (1 mentions)
Exo 3, by New England Biolabs (1 mentions)
Synergy ht plate reader, by Agilent Technologies (1 mentions)
Calcein am, by Merck Group (1 mentions)
Spark 20m, by Tecan (1 mentions)
9 protocols using screen well fda approved drug library v2
1
High-throughput Screening of FDA-Approved Drugs
2
High-throughput drug screening using ELISA
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An ELISA-based high-throughput limited drug screen was designed using an In-Cell ELISA Colorimetric Detection Kit (Thermo Fisher Scientific, Massachusetts, USA). For this, a total of 262 FDA-approved drugs were aliquoted in 384-well microplates derived from the Screenwell FDA-approved drug library V2 (Enzo Life Sciences—Cederlane—Ontario, Canada). Each drug dose from the initial drug screen was based on doses used in the clinic by patients as listed in the Drug Bank database66 (link). C2C12 myoblasts (American Type Culture Collection, Virginia, US, CRL-1772TM) were grown in each well and treated with these drugs or vehicle control for 24 h. Following treatment, antibodies targeting eEF1A2 (1:1000, provided by Dr. Abbott) or utrophin A (1:500; Novocastra, Leica biosystems, Concord, ON, Canada, NCL-DRP2) and HRP-conjugated IgG secondary antibodies (Jackson Immuno Research, Bar Harbor, USA, 111-035-033 and AP124P) were used to detect protein expression levels. Absorbance levels were determined with a Synergy H1 microplate reader. Note that the absorbance levels are standardized to total cell number by using a whole-cell stain in order to control for variation in cell proliferation.
3
Identification of Foldlin, a Novel HSP70 Inducer
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Foldlin was identified in a screen comprising approximately 900,000 small molecules that induce an HSP70 response in HeLa cells38. The compound was resynthesized by EcoSynth (Oostende, Belgium). See Supplementary Information on the synthesis and quality control in Supplementary Figure S1 . The inactive molecule was kindly provided by Proteostasis Therapeutics Inc (Boston, MA, USA). The SCREEN-WELL® FDA approved drug library V2 was obtained from Enzo life sciences (Farmingdale, NY, USA). ML346 (HY-18669) and KRIBB11 (HY-100872) were obtained from MedChemExpress (Monmouth Junction, NJ, USA).
4
Screening Library Compound Preparation
5
Comprehensive Drug Library Acquisition
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The SCREENWELL FDA‐approved drug library V2 containing 741 compounds was purchased from Enzo Life Sciences (Hayashi Kasei Co., Ltd.), and the International Drug Collection (IDC) containing 311 compounds was purchased from MicroSource Discovery Systems, Inc. (Namiki Shoji Co., Ltd.). Aldosterone (A9477), DHEA (D4000), dehydroisoandrosterone 3‐sulfate (D5297), and Reichstein's substance S (also called 11‐deoxycortisol; R0500) were obtained from Sigma. Androstenedione (A0845), cholesterol (C0318), corticosterone (C0388), estriol (E0218), hydrocortisone (H0533), and stanolone (also known as dihydrotestosterone; A0462) were purchased from Tokyo Chemical Industry (TCI). 17‐α hydroxypregnenolone (SC223186) and 21‐hydroxyprogesterone (also called deoxycorticosterone; SC231274) were obtained from Santa Cruz Biotechnology. Tyrosine kinase inhibitors imatinib mesylate (sc‐202180), nilotinib (sc‐202245), dasatinib (sc‐358114), bosutinib (sc‐202084), bafetinib (sc‐503249), and ponatinib (sc‐362710) were purchased from Santa Cruz Biotechnology, Inc. Chemical stocks (10 mM in DMSO or ethanol for cholesterol) were either obtained from the manufacturer (chemical libraries) or prepared (individual chemicals) and stored at −20°C.
6
High-Throughput Screening of FDA-Approved Drugs
7
Screening FDA Drugs for Wnt Modulators
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The 6XTCF/LEF-miniP:dGFP transgenic zebrafish line was to screen a library of FDA-approved drugs (Enzo Screen-Well® FDA Approved Drug Library V2, NY, USA, version 1.4., BML-2843–0100, Lot No. 06051910 A) for modulators of the Wnt/β-catenin/TCF-LEF signaling axis modulators. Screening was carried out as previously described [39 (link),40 (link)]. Briefly, at 24 h post-fertilization (hpf), zebrafish larvae showing similar GFP fluorescence in the tail fin were selected for use. The larvae were dechorionated and placed individually into a 96-well round-bottomed plate with 148.5 μl of 1X E3 media. The drug library was diluted to 100 μM in E3 water immediately before use and drugs were tested at a final concentration of 1 μM by adding 1.5 μl of the 100 μM diluted drug stock to each well. An equivalent volume of DMSO was used as the control. Larvae were incubated for 48hrs at 28 °C.
8
Screening FDA-Approved Drug Library for Cytotoxicity
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The Screen-Well® FDA Approved Drug Library V2 (BML-2843-0100, Enzo Life Sciences, Farmingdale, NY, USA) was used for the screening. This compound library included 774 compounds approved by the US Food and Drug Administration (FDA).
Cells (2 × 104 cells/well in 100 μL medium) were seeded into 96-well plates and grown for 24 h. The compounds of the library were transferred to the plates with a Tecan Freedom EVO liquid handling robot to a final concentration of 10 μM followed by a 24-h incubation. Then, the cells were stained by adding 50 μL of Calcein-AM (#17783, Sigma, St. Luois, MO, USA) solution at a final concentration of 1 μM. After incubating cells for 1 h at 37 °C, the fluorescent signal was measured with a Tecan Spark 20M (Tecan, Männedorf, Switzerland) multimode reader (Ex/Em = 485/530 nm). Viability was expressed as a percentage of the untreated control.
To validate the selected cytotoxic hits, cells (2 × 104 cells/well in 100 μL medium) were seeded into 96-well plates and grown for 24 h. Hit compounds were purchased from different companies, as summarized inSupplementary Table S4 . A concentration series was prepared to identify the lowest effective concentration of the compounds. After 24 h of treatment, Calcein-AM assays were used to determine cell viability, as described above.
Cells (2 × 104 cells/well in 100 μL medium) were seeded into 96-well plates and grown for 24 h. The compounds of the library were transferred to the plates with a Tecan Freedom EVO liquid handling robot to a final concentration of 10 μM followed by a 24-h incubation. Then, the cells were stained by adding 50 μL of Calcein-AM (#17783, Sigma, St. Luois, MO, USA) solution at a final concentration of 1 μM. After incubating cells for 1 h at 37 °C, the fluorescent signal was measured with a Tecan Spark 20M (Tecan, Männedorf, Switzerland) multimode reader (Ex/Em = 485/530 nm). Viability was expressed as a percentage of the untreated control.
To validate the selected cytotoxic hits, cells (2 × 104 cells/well in 100 μL medium) were seeded into 96-well plates and grown for 24 h. Hit compounds were purchased from different companies, as summarized in
9
FDA-approved Drug Library Screening
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The FDA-approved drug library (SCREEN-WELL FDA-approved drug library V2) was purchased from Enzo Life Sciences. HOC313-LM cells were seeded into 96-well plates (3.5 Â 10 4 cells/well). The next day, each drug (766 drugs, 1 mmol/L) was added to each well. After 24 hours, cells were stained with crystal violet solution. After that, stained cells were dissolved in 2% SDS, and their density was measured by an absorption spectrometer (560 nm).
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