| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
| 10mg | |||
| Other Sizes |
| Targets |
Targets: IDO1 (indoleamine 2,3-dioxygenase 1, IC50 = 28 nM) and IDO2 (indoleamine 2,3-dioxygenase 2, IC50 = 144 nM). Both IDO1 and IDO2 are immunosuppressive enzymes that catalyze the degradation of tryptophan to kynurenine, creating an immunosuppressive tumor microenvironment. Dual inhibition restores anti-tumor immune responses by preventing T cell anergy and promoting effector T cell function.
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| ln Vitro |
In vitro, IDO1/2-IN-1 HCl potently inhibits IDO1 and IDO2 with IC50s of 28 nM and 144 nM, respectively. It is the first potent dual inhibitor of IDO1 and IDO2. The compound exhibits antitumor activity in cellular assays by restoring T cell proliferation and function in the presence of immunosuppressive tumor cells. It has an EC50 in the low nanomolar to low micromolar range in cell-based functional assays.
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| ln Vivo |
In vivo, IDO1/2-IN-1 HCl is orally active and exhibits antitumor effects in syngeneic mouse tumor models. It reduces tumor growth, increases tumor-infiltrating CD8+ T cells, and decreases regulatory T cells (Tregs) in the tumor microenvironment. The compound has been shown to enhance the efficacy of immune checkpoint inhibitors (anti-PD-1/PD-L1). Detailed efficacy studies have been reported in the patent literature.
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| Enzyme Assay |
For cell-free IDO1/IDO2 enzyme inhibition assays: recombinant human IDO1 or IDO2 enzyme (50-100 ng) is incubated with varying concentrations of IDO1/2-IN-1 HCl (0-10 uM), L-tryptophan (100-200 uM), and methylene blue (10 uM) or ascorbic acid/catalase system in 100 uL assay buffer (50 mM potassium phosphate buffer pH 6.5, 20 mM ascorbic acid, 20 ug/mL catalase) at 37degC for 30-60 min. The reaction is stopped by adding trichloroacetic acid (30% final) and heated at 50-65degC for 20-30 min. After centrifugation, the supernatant is mixed with Ehrlich's reagent (p-dimethylaminobenzaldehyde in acetic acid), and absorbance is measured at 480-490 nm. IC50 values (28 nM for IDO1, 144 nM for IDO2) are calculated from dose-response curves.
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| Cell Assay |
For cell-based assays: IDO1/IDO2-expressing cells (e.g., HeLa cells stimulated with IFN-gamma, or IDO1/IDO2-transfected HEK293 cells) are seeded in 96-well plates. Cells are treated with IDO1/2-IN-1 HCl (0.1-100 uM) for 24-48 h. Supernatants are collected and mixed with Ehrlich's reagent to quantify kynurenine production. Alternatively, tryptophan depletion is measured by HPLC or LC-MS. T cell proliferation assays: human PBMCs or purified T cells are co-cultured with IDO1-expressing tumor cells in the presence of IDO1/2-IN-1 HCl, and T cell proliferation is measured by CFSE dilution or 3H-thymidine incorporation.
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| Animal Protocol |
For in vivo animal studies: syngeneic mouse tumor models (e.g., B16 melanoma, CT26 colon cancer, 4T1 breast cancer) are established in immunocompetent mice. IDO1/2-IN-1 HCl is formulated in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline and administered orally at doses of 30-100 mg/kg once or twice daily for 2-4 weeks. Tumor volume is measured by calipers. Plasma kynurenine/tryptophan ratios are measured by LC-MS. Tumor tissues are harvested for analysis of T cell infiltration (CD8+ T cells, Tregs) by flow cytometry and immunohistochemistry. Combination studies with anti-PD-1 or anti-CTLA-4 antibodies are performed. The compound exhibits antitumor effects and enhances the efficacy of immune checkpoint inhibitors.
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| ADME/Pharmacokinetics |
PK properties of IDO1/2-IN-1 HCl: In rodents after oral administration (30-100 mg/kg), the compound shows good oral bioavailability (30-60%), Tmax 1-2 h, plasma half-life 4-8 h. The compound (MW 521.73) is soluble in DMSO (100 mg/mL). In vivo formulation: 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline. Plasma protein binding is moderate (80-90%). Metabolism is primarily via CYP450 enzymes (CYP3A4). The compound has good tissue distribution, including penetration into tumor tissues.
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| Toxicity/Toxicokinetics |
No toxicity data have been reported specifically for IDO1/2-IN-1 HCl. IDO1 inhibitors in general have a favorable safety profile, with minimal toxicity observed in clinical trials. IDO1/IDO2 dual inhibition may have increased risk of autoimmune-like adverse effects due to enhanced immune activation. Potential adverse effects may include immune-related toxicities (rash, colitis, pneumonitis, hepatitis). The compound is for research use only and not for human consumption.
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| References | |
| Additional Infomation |
IDO1/2-IN-1 HCl is a research compound not yet approved for clinical use. It is the first potent dual IDO1/IDO2 inhibitor and a valuable tool for studying the role of tryptophan metabolism in tumor immune evasion. It has potential applications in cancer immunotherapy, as a single agent or in combination with immune checkpoint inhibitors (anti-PD-1/PD-L1, anti-CTLA-4), for treating various cancers including melanoma, lung cancer, breast cancer, colon cancer, and ovarian cancer. It can also be used to study IDO2 biology, which is less characterized than IDO1.
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| Molecular Formula |
C16H19BRCLFN8O4
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|---|---|
| Molecular Weight |
521.73
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| Exact Mass |
520.038
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| CAS # |
2310286-60-1
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| Related CAS # |
IDO1/2-IN-1;2310286-45-2
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| PubChem CID |
141755880
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
11
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| Rotatable Bond Count |
11
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| Heavy Atom Count |
31
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| Complexity |
553
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=CC(=C(C=C1N=C(C2=NON=C2NCCN3C=C(N=N3)COCCO)NO)Br)F.Cl
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| InChi Key |
OFKAFAFCZQUHPC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H18BrFN8O4.ClH/c17-12-7-10(1-2-13(12)18)20-16(22-28)14-15(24-30-23-14)19-3-4-26-8-11(21-25-26)9-29-6-5-27;/h1-2,7-8,27-28H,3-6,9H2,(H,19,24)(H,20,22);1H
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| Chemical Name |
N'-(3-bromo-4-fluorophenyl)-N-hydroxy-4-[2-[4-(2-hydroxyethoxymethyl)triazol-1-yl]ethylamino]-1,2,5-oxadiazole-3-carboximidamide;hydrochloride
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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: ~100 mg/mL (~191.7 mM; with ultrasonication)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (4.79 mM) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (4.79 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of 20% SBE-β-CD saline and mix well. 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. Solubility in Formulation 3: 2.5 mg/mL (4.79 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), Clear solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of DMSO stock solution (25.0 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9167 mL | 9.5835 mL | 19.1670 mL | |
| 5 mM | 0.3833 mL | 1.9167 mL | 3.8334 mL | |
| 10 mM | 0.1917 mL | 0.9584 mL | 1.9167 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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