NLRP3-IN-15 is a potent and specific inhibitor of the NLRP3 inflammasome (KD: 5.87 μM). NLRP3-IN-15 inhibits IL-1β release with IC50 of 0.131 μM. NLRP3-IN-15 may be utilized in inflammation research.

Physicochemical Properties

Molecular Formula	C22H19NO4
Molecular Weight	361.39
Exact Mass	361.131
CAS #	2767369-71-9
PubChem CID	168006890
Appearance	Typically exists as solid at room temperature
LogP	2.6
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	2
Heavy Atom Count	27
Complexity	621
Defined Atom Stereocenter Count	0
InChi Key	GCYKCXXLWUWUFQ-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H19NO4/c1-12-3-2-4-14-13(12)5-6-16-18(14)20(25)21(26)19-15-7-8-23(9-10-24)11-17(15)27-22(16)19/h2-6,24H,7-11H2,1H3
Chemical Name	6-(2-hydroxyethyl)-18-methyl-3-oxa-6-azapentacyclo[11.8.0.02,10.04,9.014,19]henicosa-1(13),2(10),4(9),14,16,18,20-heptaene-11,12-dione
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	NLRP3 inflammasome
ln Vitro	By using an ELISA experiment, compound 12a, NLRP3-IN-15, suppresses the release of IL-1β with an IC50 of 0.114 μM [1]. In mouse peritoneal macrophages (PM), NLRP3-IN-15 (2 μM, 3 hours) suppresses the production of caspase-1 (p20) and IL-1β (p17) [1]. By preventing ASC oligomerization, NLRP3-IN-15 prevents the NLRP3 inflammasome complex from forming [1]. In human and mouse liver microsomes (T1/2: 693 min; Clint: 2.0 μL/min/mg), NLRP3-IN-15 demonstrates metabolic stability[1].
ln Vivo	In a mouse model of sepsis, NLRP3-IN-15 (compound 5j) (50 mg/kg; ip) shows anti-inflammatory properties[1].
Cell Assay	Western Blot Analysis[1] Cell Types: Mice peritoneal macrophages(PMs) Tested Concentrations: 2 μM Incubation Duration: 3 h Experimental Results: Inhibited the secretion of IL-1β (p17) and caspase-1 (p20), without affecting pro-IL-1β and pro-caspase-1.
Animal Protocol	Animal/Disease Models: LPS-induced inflammatory septic mouse model[1] Doses: 50 mg/kg Route of Administration: ip Experimental Results: diminished the release of IL-1β in mouse serum. Relieved thickening of the alveolar wall in lung. Animal/Disease Models: Mice[1] Doses: 20 mg/kg Route of Administration: po or ip Experimental Results: pharmacokinetic/PK profile of NLRP3-IN-15 (compound 12a). parameter dose (mg/kg) T1/2 (h) Tmax (h) F (%) PO 20 1.659 0.5 9.6 IP 20 1.807 0.167 21.2
References	[1]. Scaffold Hybrid of the Natural Product Tanshinone I with Piperidine for the Discovery of a Potent NLRP3 Inflammasome Inhibitor. J Med Chem. 2023 Feb 23;66(4):2946-2963.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.7671 mL	13.8355 mL	27.6709 mL
	5 mM	0.5534 mL	2.7671 mL	5.5342 mL
	10 mM	0.2767 mL	1.3835 mL	2.7671 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.