JAK-IN-23 is a dual (bifunctional) inhibitor of JAK/STAT and NF-κB with oral activity. JAK-IN-23 has an inhibitory activity against J AK1/2/3, with IC50s of 8.9 nM, 15 nM and 46.2 nM respectively. JAK-IN-23 has strong inhibitory activity against interferon-stimulated genes (ISG) and NF-κB pathways, with IC50s of 3.3 nM and 150.7 nM respectively. JAK-IN-23 has a strong anti~inflammatory effect and can reduce the release of a variety of pro-inflammatory factors. JAK-IN-23 may be utilized in the research/study of inflammatory bowel disease (IBD).

Physicochemical Properties

Molecular Formula	C23H22CL2N4O
Molecular Weight	441.35
CAS #	3031837-35-8
PubChem CID	165368933
Appearance	Light brown to gray solid powder
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	30
Complexity	592
Defined Atom Stereocenter Count	0
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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

ln Vitro	With IC50 values of 8.9 nM, 15 nM, and 46.2 nM, respectively, JAK-IN-23 inhibits JAK1/2/3[1]. Strong inhibitory effects against NF-KB and ISG are demonstrated by JAK-IN-23, which has IC50 values of 150.7 nM and 3.3 nM, respectively[1]. In THP1-dual cells, WB---JAK-IN-23 (0.33μM, 1μM, 3μM; 24 h) can block the proinflammatory signaling pathways of NF-κB and JAK-STAT1/3 simultaneously[1]. In THP1-dual cells stimulated by LPS, JAK-IN-23 (0.003-3 μM; 24 h) reduces the release of several proinflammatory factors, such as IL-6, IL-8, and IL-1β[1]. In LPS-induced peripheral blood mononuclear cells (PBMCs), JAK-IN-23 (0.11-3 μM; 24 h) reduces the release of multiple proinflammatory factors, such as TNF-α, IL-12, IL-10, and IFNγ[1]. JAK-IN-23 (1 μM) has inhibitory effects on the expression of genes involved in the unfolded protein response that was induced by LPS (1 μg/mL) and inhibits the expression of many inflammation-related genes induced by LPS, such as IL-1B, TNF, IL12B, and IL-23A[1].
ln Vivo	In acute enteritis models induced by dextran sulfate sodium (DSS) and 2,4,6-trinitrobenzenesulfonic acid (TNBS), JAK-IN-23 (1–5 mg/kg, oral) exhibits potent anti-inflammatory activity and restores the structural makeup of the gut microbiota[1].
Cell Assay	Western Blot Analysis[1] Cell Types: THP1-Dual Cells Tested Concentrations: 0.33μM, 1μM, 3μM Incubation Duration: 24 h Experimental Results: Inhibited p-STAT1/ 3 in a dose-dependent manner that was induced by IL-6, as well as inhibited pNF-κB p65 in a dose-dependent manner, but not on MYD88 and p-IKK α/β that was induced by LPS.
Animal Protocol	Animal/Disease Models: DSS-Induced Acute Colitis Mice Model[1] Doses: 1 mg/kg, 3 mg/kg Route of Administration: oral Experimental Results: Dramatically diminished the DAI scores (1 and 3 mg/ kg). Recovered the length of the colon (3 mg/kg). Dramatically decreased the histopathology of ulcerative colitis (1 and 3 mg/kg). Animal/Disease Models: The BALB/c mouse inflammatory bowel disease (IBD) model[1] Doses: 1 mg/kg, 5 mg/kg Route of Administration: oral Experimental Results: Dramatically improved the survival probability, had low DAI scores and effectively relieved symptoms of colitis in the TNBS-induced IBD mice model (5 mg/kg). Did not improve the survival probability and decreases the DAI score (100 mg/kg).
References	[1]. Discovery of Novel Imidazo[4,5- c]quinoline Derivatives to Treat Inflammatory Bowel Disease (IBD) by Inhibiting Multiple Proinflammatory Signaling Pathways and Restoring Intestinal Homeostasis. J Med Chem. 2022 Sep 2.

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	2.2658 mL	11.3289 mL	22.6578 mL
	5 mM	0.4532 mL	2.2658 mL	4.5316 mL
	10 mM	0.2266 mL	1.1329 mL	2.2658 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.