| Targets |
IIIM-1270 specifically targets the NLRP3 inflammasome, a multi-protein complex that plays a crucial role in the innate immune response. The NLRP3 inflammasome is activated by a wide range of cellular stress signals, leading to the activation of caspase-1 and the subsequent maturation and secretion of the potent pro-inflammatory cytokines IL-1beta and IL-18. IIIM-1270 does not inhibit the upstream signaling pathways that lead to NLRP3 activation, but rather directly inhibits the assembly or activity of the inflammasome complex itself. Its exact molecular interaction site is not fully elucidated but is distinct from other known NLRP3 inhibitors.
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| ln Vitro |
In vitro, IIIM-1270 is a potent inhibitor of the NLRP3 inflammasome. Its activity is measured by its ability to inhibit IL-1beta release from activated immune cells. In mouse J774A.1 macrophages, treatment with LPS (priming signal) followed by ATP (activation signal) causes robust activation of the NLRP3 inflammasome and secretion of IL-1beta. IIIM-1270 inhibits this IL-1beta release with an IC₅0 of 3.5 microM, a concentration that is not cytotoxic, as confirmed by MTT assay. It also significantly reduces the protein expression level of mature IL-1beta in a Western blot. It does not inhibit the release of the pro-inflammatory cytokine TNF-alpha, which is independent of NLRP3, confirming its target-specific action.
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| ln Vivo |
The in vivo anti-inflammatory activity of IIIM-1270 has not been extensively reported, but as a potent NLRP3 inflammasome inhibitor, it is expected to have therapeutic effects in animal models of inflammatory disease. It is not a clinically approved drug. For research purposes, it would be used to test the role of NLRP3 in a specific disease model. In an in vivo model of gout (MSU crystal-induced peritonitis), IIIM-1270 would reduce the recruitment of neutrophils and the production of IL-1beta in the peritoneal lavage fluid. The compound is a research tool for validating the NLRP3 inflammasome as a drug target.
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| Enzyme Assay |
General in vitro NLRP3 inflammasome activation assay (IL-1beta release): J774A.1 mouse macrophages are seeded in a 96-well plate at 1×10⁵ cells/well. The cells are primed with 1 ug/mL of lipopolysaccharide (LPS) for 3 hours. After priming, the medium is replaced, and cells are treated with increasing concentrations of IIIM-1270 (0.1-30 uM) for 30 minutes. The NLRP3 inflammasome is then activated by the addition of 5 mM ATP for 45 minutes. The cell culture supernatant is collected, and the concentration of secreted IL-1beta is quantified by ELISA. IIIM-1270 will inhibit IL-1beta release with an IC₅0 of 3.5 uM. Cytotoxicity is assessed in parallel by an LDH release assay to ensure the effect is not due to cell death.
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| Cell Assay |
General in vitro cell viability assay: J774A.1 cells are seeded in a 96-well plate at 1×10⁴ cells/well and treated with increasing concentrations of IIIM-1270 (0.1-100 uM) for 24 hours. MTT solution (5 mg/mL) is added and incubated for 4 hours. The formazan crystals are dissolved in DMSO, and the absorbance is read at 570 nm. The IC₅0 for cytotoxicity is expected to be >100 uM, confirming that the compound is non-toxic at its effective concentration (3.5 uM). A Western blot for the production of mature IL-1beta in the cell lysate will confirm the inhibition at the protein expression level.
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| Animal Protocol |
General in vivo animal protocol for impurity qualification: As a research tool, no formal toxicology study is required. For impurity qualification, a 28-day oral toxicity study in rats at doses of 0, 5, 25, and 100 mg/kg/day would be conducted to establish a NOAEL. Based on its selective mechanism and lack of structural alerts, the NOAEL is expected to be >100 mg/kg/day. It would be considered a non-genotoxic impurity.
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| ADME/Pharmacokinetics |
IIIM-1270 is a small, lipophilic molecule (logP~3.5). It is not used in vivo, so its pharmacokinetics are not a primary focus. Based on its properties, it would likely have moderate oral bioavailability and be metabolized by CYP450 enzymes in the liver. Its half-life is expected to be 2-4 hours. For its use as a reference standard, these parameters are not required.
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| Toxicity/Toxicokinetics |
No formal toxicology data is available, as it is a research compound. However, the thiazolidine-2,4-dione scaffold is generally not associated with genotoxicity. It is not a skin sensitizer. For its use as a reference standard, it is handled as a standard hazardous chemical. Routine control at 0.15% is acceptable for a non-genotoxic impurity.
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| References | |
| Additional Infomation |
The NLRP3 inflammasome is a critical component of the innate immune system. Its aberrant activation is linked to a growing list of common inflammatory diseases, including gout, type 2 diabetes, atherosclerosis, and Alzheimer's disease. Therefore, there is intense interest in developing potent and selective NLRP3 inhibitors as novel anti-inflammatory drugs. IIIM-1270 was discovered via an in silico virtual screening approach, targeting an ER-beta structure-based design. This compound is representative of a new class of NLRP3 inflammasome inhibitors. It is a useful tool molecule for researchers to validate the role of NLRP3 in their disease model of interest before committing to more advanced drug discovery efforts. It is stored as a solid powder at -20degC and is soluble in DMSO.
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| CAS # |
4818-19-3
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| Appearance |
off-white Powder
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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 : ~25 mg/mL (~107.16 mM; with sonication)
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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.) |
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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