Gc376

Manufactured by Targetmol

Sourced in United States

GC376 is a small molecule compound. It functions as a protease inhibitor.

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Lab products found in correlation

8 protocols using gc376

Antiviral Activity of Flavonoids Against FIPV

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Crandell–Rees feline kidney cells were seeded into a 96-well plate at 1.8 × 10⁴ cells/well and cultured in MEM containing 7% FBS. Pre-compound, attachment, penetration, post-viral entry, and virucidal assays were performed based on each step of viral infection. All assays were performed using various concentrations of flavonoids: 100, 50, 25, 10, 1, and 0.1 μM/well with FIPV-infected CRFK cells. For the antiviral activity assay, FIPV was used at 50 TCID₅₀/well in all experiments. Crandell–Rees feline kidney cells were inoculated with FIPV and treated with 0.1% DMSO as vehicle control. GC376 (TargetMol®, Wellesley Hills, MA, USA) was used as the FIPV-specific drug control [17 (link), 18 (link)]. Three experiments were performed in duplicate wells for each compound. The conditions of the antiviral activity assay are shown in Figure-1a.

Triratapiban C., Lueangaramkul V., Phecharat N., Pantanam A., Lekcharoensuk P, & Theerawatanasirikul S. (2023). First study on in vitro antiviral and virucidal effects of flavonoids against feline infectious peritonitis virus at the early stage of infection. Veterinary World, 16(3), 618-630.

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SARS-CoV-2 Protease Inhibitor Screening

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RDV (Gilead, Foster City, CA, USA) and GC376 (TargetMol, Boston, MA, USA) were used as reference compounds to assess the anti-SARS-CoV-2 in vitro activity or viral 3CL^pro protease activity, respectively. The CDK inhibitors LDC4297 (Lead Discovery Center, GmbH, Dortmund, Germany), SNS 032 (Tocris, Bristol, United Kingdom) and R25 (also termed alsterpaullone, GPC Biotech AG, Martinsried, Germany) were obtained from indicated sources.

Hahn F., Häge S., Herrmann A., Wangen C., Kicuntod J., Jungnickl D., Tillmanns J., Müller R., Fraedrich K., Überla K., Kohlhof H., Ensser A, & Marschall M. (2021). Methodological Development of a Multi-Readout Assay for the Assessment of Antiviral Drugs against SARS-CoV-2. Pathogens, 10(9), 1076.

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Evaluation of Anti-SARS-CoV-2 Activity

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RDV (Gilead, Foster City, CA, USA) and GC376 (T5188, TargetMol, Boston, MA, USA) were used as the reference compounds to determine the anti-SARS-CoV-2 activity for recSARS-CoV-2. The CaCo-2 cells were treated with 2-fold serial dilutions of either RDV (20 nM–0.16 nM) or GC376 (80 nM–0.63 nM) and infected with different SARS-CoV-2 strains at an MOI of 0.005. At 30 hpi, the cells were fixed and analyzed. The infection rates were assessed by immunostaining of the viral spike and reporter expression, respectively. The percentage of viral replication was determined relative to the solvent-treated cells. The 50% effective concentration (EC₅₀) was assessed by nonlinear four-parameter curve fitting using GraphPad Prism 6 (GraphPad Software, San Diego, CA, USA).

Herrmann A., Jungnickl D., Cordsmeier A., Peter A.S., Überla K, & Ensser A. (2021). Cloning of a Passage-Free SARS-CoV-2 Genome and Mutagenesis Using Red Recombination. International Journal of Molecular Sciences, 22(19), 10188.

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Antcins and Terpene Compounds Bioassay

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GC376 (≥ 98%) (Cat # T5188) was purchased from TargetMol (Washington Street, Wellesley Hills, MA, USA). High glucose Dulbecco's Modified Eagle's Medium (DMEM)/high glucose containing 4 mM L-Glutamine, 4.5 g/L glucose (Cat # AE29444172) without sodium pyruvate, penicillin–streptomycin antibiotic solutions, and fetal bovine serum (FBS) were obtained from Hyclone/G.E. Healthcare Life Sciences (Logan, UT, USA). Antcins (A, B, C, H, I, and M) were isolated from fruiting bodies of T. camphoratus and A. salmonea, as described previously^{16 (link),55 (link)}, and purchased from the R&D Center of Taiwan Leader Biotechnology Corp. (Taichung, Taiwan) and purified using HPLC and FT-NMR analysis. GC-grade citronellol (Cat # C0370) and Limonene (Cat # L0132) were obtained from Tokyo Chemical Industry Co., Ltd (Toshima, Tokyo, Japan).

Dakpa G., Kumar K.J., Nelen J., Pérez-Sánchez H, & Wang S.Y. (2023). Antcin-B, a phytosterol-like compound from Taiwanofungus camphoratus inhibits SARS-CoV-2 3-chymotrypsin-like protease (3CLPro) activity in silico and in vitro. Scientific Reports, 13, 17106.

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Synthesis and Procurement of Compounds

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The protocol for TF27 synthesis has been detailed previously [10 (link),20 (link)]. A new batch was synthesized by Vichem Chemie Research Ltd. (Budapest, Hungary). Artesunate (ART) was purchased from Saokim Ltd. (Hanoi, Vietnam). EIDD-1931 was kindly provided by Immunic AG, Gräfelfing, Germany. GC376 was purchased from TargetMol (Boston, MA, USA). Stock aliquots were prepared in pure DMSO (Sigma, St. Louis, MO, USA) at 10 mM and stored at −20 °C.

Hahn F., Wangen C., Häge S., Herrmann L., Herrmann A., Tsogoeva S.B, & Marschall M. (2022). The Trimeric Artesunate Analog TF27, a Broadly Acting Anti-Infective Model Drug, Exerts Pronounced Anti-SARS-CoV-2 Activity Spanning Variants and Host Cell Types. Pharmaceutics, 15(1), 115.

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Biochemical Characterization of Inhibitors

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The natural product library, GC-376, and 7-methoxycoumarin-4-acetic acid (MCA) were purchased from TargetMol (Shanghai, China), and the purity was more than 97.0% in mass spectrometry analysis. Dieckol was commercially provided by Yuanye Biotech (Shanghai, China). All tested compounds were dissolved in DMSO at a final concentration of 20 mM, and stored at − 20 °C before use. The FRET fluorogenic substrate (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH₂), FP tracer (FITC-AVLQSGFRKK-Biotin) and FITC-AVLQ peptide were chemically synthesized (GL Biochem Shanghai Co. Ltd., Shanghai, China), and the purity was more than 95.0%.

Yan G., Li D., Lin Y., Fu Z., Qi H., Liu X., Zhang J., Si S, & Chen Y. (2021). Development of a simple and miniaturized sandwich-like fluorescence polarization assay for rapid screening of SARS-CoV-2 main protease inhibitors. Cell & Bioscience, 11, 199.

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Evaluation of Protease and Proteasome Inhibitors

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Saquinavir (Cat no. HY-17007), Ritonavir (Cat no. HY-90001), Indinavir (Cat no. HY-B0689), Nelfinavir Mesylate (Cat no. HY-15287A), Amprenavir (Cat no. HY-17430), Lopinavir (Cat no. HY-14588), Atazanavir sulfate (Cat no. HY-17367A), Fosamprenavir (Cat no. HY-78726), Tipranavir (Cat no. HY-15148), Darunavir (Cat no. HY-17040), Boceprevir (Cat no. HY-10237), Telaprevir (Cat no. HY-10235), Simeprevir (Cat no. HY-10241), Asunaprevir (Cat no. HY-14434), Grazoprevir (Cat no. HY-15298), Carfilzomib (Cat no. HY-10455), and Bortezomib (Cat no. HY-10227) were purchased from MedChemExpress. GC376 (Cat no. T5188) were purchased from TargetMol.

Fu L., Ye F., Feng Y., Yu F., Wang Q., Wu Y., Zhao C., Sun H., Huang B., Niu P., Song H., Shi Y., Li X., Tan W., Qi J, & Gao G.F. (2020). Both Boceprevir and GC376 efficaciously inhibit SARS-CoV-2 by targeting its main protease. Nature Communications, 11, 4417.

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Protocol for Inhibitor Evaluation

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Geraniin, nirmatrelvir (PF-07321332, PF-332), and GC-376 were purchased from TargetMol (Shanghai, China). The FRET substrate (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2), MCA-AVLQ peptide, and FP probe (FITC-AVLQSGFRKK-Biotin) were chemically synthesized (GL Biochem Shanghai Co. Ltd., Shanghai, China) , and the purity was more than 95.0%.

Zhang R., Yan H., Zhou J., Liu X, & Chen Y. (2024). Invalidation of geraniin as a potential inhibitor against SARS-CoV-2 main protease. Natural product research, 38(12).

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