Table 1.
Identified chemical inhibitors against Diguanylate Cyclases (DGCs).
| Name of chemical | Mode of action | Screening and DGC activity assay methods | Chemical library used for screening | References |
|---|---|---|---|---|
| Natural molecules | ||||
| Glycosylated Triterpenoid Saponin | Non-competitive inhibition | In vitro enzyme activity assay analyzed by TLC (thin layer chromatograph) followed by autoradiography | Plant extract | Ohana et al., 1998 |
| c-di-GMP analogs | ||||
| 2′-F-c-di-GMP | Non-competitive inhibition binding to the DGC I-site | In vitro enzyme activity assay analyzed by TLC followed by autoradiography | Three c-di-GMP analogs chemically synthesized | Zhou et al., 2013 |
| c-di-Inosinylic Acid | Non-competitive inhibition binding to the DGC I-site | In vitro enzyme activity assay analyzed by HPLC (high performance liquid chromatography) | Five c-di-GMP analogs chemically synthesized | Ching et al., 2010 |
| Triazole-Linked Analog DCI058 | Non-competitive inhibition binding to the DGC I-site | In vitro enzyme activity assay analyzed by circular dichroism (CD) spectroscopy | 16 c-di-GMP analogs chemically synthesized | Fernicola et al., 2015 |
| GTP analogs | ||||
| MANT-GTP and MANT-GTPγS | Unknown, probably competitive inhibition | In vitro enzyme activity assay analyzed by HPLC-MS/MS | 8 NTP derivatives | Spangler et al., 2011 |
| TNP-GTP | Unknown, probably competitive inhibition | In vitro enzyme activity assay analyzed by HPLC-MS/MS | 8 NTP derivatives | Spangler et al., 2011 |
| Small synthetic molecules | ||||
| Ebselen | Cysteine residue modification near the I-site | DRaCALA (Differential Radial Capillary Action of Ligand Assay) | NIH clinical collection 1 (NCC1) library | Lieberman et al., 2014 |
| Catechol-containing Sulfonohydrazide compounds | Competitive inhibition of active site | In silico (3-D Pharmacophore) prediction followed by in vitro enzyme activity assay | The purchasable subset of the ZINC database (~2.3 × 107 compounds) | Fernicola et al., 2016 |
| Sulfasalazine | Competitive inhibition of active site | In silico prediction with OpenEye Scientific Software followed by in vitro enzyme activity assay | 1,500 FDA-approved drugs in the DrugBank database | Wiggers et al., 2018 |
| Eprosartan | Competitive inhibition of active site | In silico prediction with OpenEye Scientific Software followed by in vitro enzyme activity assay | 1,500 FDA-approved drugs in the DrugBank database | Wiggers et al., 2018 |
| N-(4-anilinophenyl)benzamide (aka. DI-3) | Unknown | Screening with an in vivo luciferase reporter assay followed by an in vitro enzyme activity assay | 66,000 compounds/natural product extracts at the Center for Chemical Genomics at the University of Michigan | Sambanthamoorthy et al., 2012 |
| [2- oxo-2-(2-oxopyrrolidin-1-yl)ethyl] 1,3-benzothiazole-6-carboxylate | Non-competitive inhibition binding to the DGC I-site | FRET (Foster resonance energy transfer)-based in vitro enzyme activity assay | 27,502 commercially available small molecules | Christen et al., 2019 |
| 4-(2,5-dimethylphenoxy)-N-(4-morpholin-4-ylphenyl)butanamide | Non-competitive inhibition binding to the DGC I-site | FRET (Foster resonance energy transfer)-based in vitro enzyme activity assay | 27,502 commercially available small molecules derived from chemical libraries at the Institute of Chemical Biology at Harvard University | Christen et al., 2019 |
| N′-((1E)-{4-ethoxy-3-[(8-oxo-1,5,6,8-tetrahydro-2H-1,5-methanopyrido[1,2-a] [1,5]diazocin-3(4H)-yl)methyl]phenyl}methylene)-3,4,5-trihydroxybenzohydrazide (aka. LP-3134) | Competitive inhibition of active site | In silico (3-D Pharmacophore) prediction followed by in vitro enzyme activity assay | A database of commercially available compounds | Sambanthamoorthy et al., 2014 |