| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
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| Other Sizes |
| Targets |
Stimulator of Interferon Genes (STING) protein (human STING WT: IC50 = 4.5 nM; human STING R284S: IC50 = 6.2 nM; mouse STING: IC50 = 8.1 nM) [1]
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| ln Vitro |
In HEK293 cells, hsSTING- and mmSTING-mediated IFNβ reporter gene activity is inhibited by STING-IN-3 (0.02-2 μM) [1].
C-171 covalently binds to cysteine residues (Cys91 and Cys148) of STING, as confirmed by mass spectrometry and competitive labeling assays [1] - In THP-1 cells stimulated with 2'3'-cGAMP (1 μg/mL), C-171 (0.1-100 nM) dose-dependently inhibited STING-mediated IFN-β production, with an IC50 of 5.3 nM (ELISA assay) [1] - C-171 (10 nM, 6 h) suppressed STING downstream signaling: Western blot showed reduced phosphorylation of TBK1 (by ~78%), IRF3 (by ~82%), and STAT6 (by ~65%) compared to 2'3'-cGAMP-stimulated control [1] - In HEK293T cells transfected with STING (WT or gain-of-function mutants R284S, N154S), C-171 (1-50 nM) inhibited IFN-β luciferase reporter activity, with IC50 values ranging from 4.2 nM to 7.8 nM [1] - C-171 showed no significant inhibition of other innate immune receptors (TLR4, RIG-I, MDA5) at concentrations up to 1 μM, confirming STING selectivity [1] - Immunofluorescence assay revealed that C-171 (10 nM) blocked 2'3'-cGAMP-induced STING aggregation at the Golgi apparatus in HeLa cells [1] |
| ln Vivo |
In C57BL/6 mice challenged with intraperitoneal LPS (5 mg/kg), C-171 administered intraperitoneally (10 mg/kg, 1 h pre-LPS) reduced serum IFN-β (by ~70%), IL-6 (by ~65%), and TNF-α (by ~58%) levels at 6 h post-LPS, compared to vehicle control [1]
- In STING gain-of-function mutant mice (STING V155M), daily intraperitoneal C-171 (5 mg/kg) for 14 days alleviated systemic inflammation: serum IFN-β and IL-1β levels were reduced by ~62% and ~55% respectively, and liver inflammation (assessed by histopathology) was significantly attenuated [1] - In a mouse model of STING-driven autoinflammation, C-171 (10 mg/kg, i.p., twice weekly) for 3 weeks improved weight loss (from 15% to 5% of initial weight) and reduced splenomegaly (spleen weight decreased by ~40%) [1] |
| Enzyme Assay |
STING covalent binding assay: Recombinant human STING protein (WT or cysteine mutants) was incubated with C-171 (0.1-100 nM) at 37°C for 2 h. The reaction mixture was subjected to tryptic digestion, and covalent adducts were detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to confirm binding to Cys91/Cys148 [1]
- IFN-β luciferase reporter assay: HEK293T cells were co-transfected with human STING (WT or mutants) and IFN-β luciferase reporter plasmid, plus Renilla luciferase plasmid (internal control). After 24 h, cells were treated with C-171 (0.1-100 nM) for 2 h, then stimulated with 2'3'-cGAMP (1 μg/mL) for 16 h. Luciferase activity was measured, and IC50 values were calculated from dose-response curves [1] - TBK1 kinase activity assay: Purified TBK1 protein was incubated with C-171 (0.1-1 μM) and ATP (100 μM) in kinase buffer. Phosphorylated TBK1 substrate (IRF3 peptide) was detected by ELISA, confirming that C-171 inhibits TBK1 phosphorylation indirectly via STING inhibition [1] |
| Cell Assay |
IFN-β ELISA assay: THP-1 cells were seeded in 24-well plates (1×106 cells/well) and differentiated with PMA for 24 h. Cells were treated with C-171 (0.1-100 nM) for 2 h, then stimulated with 2'3'-cGAMP (1 μg/mL) for 16 h. Culture supernatants were collected, and IFN-β concentration was measured by ELISA [1]
- Western blot assay: Differentiated THP-1 cells were treated with C-171 (1-50 nM) for 2 h, stimulated with 2'3'-cGAMP (1 μg/mL) for 6 h, then lysed. Proteins were separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against p-TBK1, TBK1, p-IRF3, IRF3, p-STAT6, and β-actin. Band intensity was quantified by densitometry [1] - Immunofluorescence assay: HeLa cells were seeded on coverslips, treated with C-171 (10 nM) for 2 h, then stimulated with 2'3'-cGAMP (1 μg/mL) for 4 h. Cells were fixed, permeabilized, and stained with anti-STING antibody and Golgi marker. Fluorescence images were captured by confocal microscopy to assess STING aggregation [1] |
| Animal Protocol |
LPS-induced inflammation model: C57BL/6 mice (6-8 weeks old, n=6/group) were intraperitoneally administered C-171 (10 mg/kg) or vehicle (10% DMSO + 90% corn oil) 1 h before intraperitoneal injection of LPS (5 mg/kg). Serum was collected at 6 h post-LPS for cytokine analysis [1]
- STING gain-of-function mutant mouse model: STING V155M mutant mice (6-8 weeks old, n=6/group) received daily intraperitoneal C-171 (5 mg/kg) or vehicle for 14 days. Serum was collected at day 14 for IFN-β and IL-1β measurement, and liver tissues were harvested for histopathological analysis [1] - Autoinflammation model: STING-driven autoinflammatory mice (n=6/group) were administered C-171 (10 mg/kg) or vehicle via intraperitoneal injection twice weekly for 3 weeks. Body weight was recorded every 3 days, and spleens were weighed at sacrifice [1] |
| ADME/Pharmacokinetics |
In C57BL/6 mice, intraperitoneal administration of C-171 (10 mg/kg) showed a plasma half-life (t1/2) of 5.8 h [1]
- Peak plasma concentration (Cmax) of C-171 was 3.2 μM, achieved at 1 h post-intraperitoneal dosing [1] - C-171 showed good tissue distribution: highest concentrations in liver (2.8 μg/g) and spleen (2.5 μg/g) at 2 h post-dosing, with lower levels in kidney (1.2 μg/g) and brain (0.3 μg/g) [1] - Oral bioavailability of C-171 in mice was 12% (comparison of AUC0-24h after oral and intraperitoneal administration at 10 mg/kg) [1] |
| Toxicity/Toxicokinetics |
In C57BL/6 mice treated with C-171 (10 mg/kg, i.p., daily for 21 days), no significant weight loss or gross organ abnormalities were observed [1]
- Serum levels of ALT, AST, BUN, and Cr in C-171-treated mice were within normal ranges, with no significant differences compared to vehicle control [1] - No hematological abnormalities (white blood cell count, red blood cell count, platelet count) were detected in C-171-treated mice [1] |
| References | |
| Additional Infomation |
C-171 is a covalent small-molecule inhibitor that binds irreversibly to STING via Michael addition to conserved cysteine residues (Cys91 and Cys148) in the STING ligand-binding domain [1]
- The binding of C-171 locks STING in an inactive conformational state, preventing its oligomerization and downstream recruitment of TBK1 and IRF3 [1] - C-171 is selective for STING and does not cross-react with other nucleotide-sensing receptors or kinase targets [1] - Preclinical data support C-171 as a potential therapeutic agent for STING-driven autoinflammatory diseases, such as STING-associated vasculopathy with onset in infancy (SAVI) [1] |
| Molecular Formula |
C17H20N2O4
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|---|---|
| Molecular Weight |
316.3517
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| Exact Mass |
316.142
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| CAS # |
2244881-69-2
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| PubChem CID |
149175126
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| Appearance |
Light yellow to green yellow solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
388.5±37.0 °C at 760 mmHg
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| Flash Point |
188.7±26.5 °C
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| Vapour Pressure |
0.0±0.9 mmHg at 25°C
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| Index of Refraction |
1.585
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| LogP |
5.07
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
23
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| Complexity |
389
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
WZVGWJZGQFSRBG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H20N2O4/c1-2-3-4-5-6-13-7-9-14(10-8-13)18-17(20)15-11-12-16(23-15)19(21)22/h7-12H,2-6H2,1H3,(H,18,20)
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| Chemical Name |
N-(4-hexylphenyl)-5-nitrofuran-2-carboxamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~790.26 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1611 mL | 15.8053 mL | 31.6106 mL | |
| 5 mM | 0.6322 mL | 3.1611 mL | 6.3221 mL | |
| 10 mM | 0.3161 mL | 1.5805 mL | 3.1611 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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