| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg | |||
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| Targets |
IDO1 (Indoleamine 2,3-Dioxygenase 1). (Rac)-IDO1-IN-5 selectively binds to the heme-deficient apo-IDO1, preventing the conversion of tryptophan to kynurenine. This unique binding mode does not require the presence of heme in the active site. By inhibiting IDO1 activity, the racemate reduces kynurenine production, reverses tryptophan depletion, and alleviates IDO1-mediated immune suppression.
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| ln Vitro |
(Rac)-IDO1-IN-5 is a potent inhibitor of IDO1 activity in cell-free enzyme assays, with an IC50 similar to that of the active S-enantiomer (approximately 100-500 nM, exact IC50 depends on assay conditions). It is highly selective for IDO1 over IDO2, TDO, and other heme-containing enzymes. The racemate shows brain-penetrant properties, making it suitable for studying CNS applications of IDO1 inhibition.
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| ln Vivo |
In mouse tumor models (e.g., B16 melanoma, CT26 colon carcinoma), oral administration of (Rac)-IDO1-IN-5 (10-100 mg/kg) reduces intratumoral kynurenine levels, increases CD8+ T-cell infiltration, and suppresses tumor growth, particularly when combined with immune checkpoint inhibitors (e.g., anti-PD-1, anti-CTLA-4). These effects are dose-dependent and correlate with plasma and tumor drug concentrations, confirming target engagement.
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| Enzyme Assay |
Cell-free fluorogenic enzyme assays for IDO1 activity are performed using purified recombinant human IDO1 protein. IDO1 (50 nM) is pre-incubated with varying concentrations of (Rac)-IDO1-IN-5 (0.1 nM to 10 uM) in 50 mM potassium phosphate buffer (pH 6.5) containing 20 mM ascorbic acid, 20 uM methylene blue, and 0.5 mg/mL catalase for 15 minutes at 37degC. The reaction is initiated by adding L-tryptophan (200 uM) and incubated for 30 minutes at 37degC. The reaction is terminated with 30% trichloroacetic acid and heated at 65degC for 15 minutes. After centrifugation, the supernatant is mixed with Ehrlich's reagent, and absorbance at 480 nm is measured. The IC50 is calculated from the inhibition of kynurenine production relative to vehicle-treated control wells.
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| Cell Assay |
HT-29 human colon adenocarcinoma cells or HeLa cells are cultured in RPMI-1640 or DMEM with 10% FBS. Cells are seeded in 96-well plates (30,000-50,000 cells/well) and treated with IFN-gamma (50-100 ng/mL) to induce IDO1 expression. Varying concentrations of (Rac)-IDO1-IN-5 (0.1 nM to 10 uM) are added, and cells are incubated for 24-48 hours at 37degC, 5% CO2. The culture supernatant is collected, and kynurenine concentration is measured by HPLC-UV (360 nm detection) or LC-MS/MS. The percent inhibition of cellular IDO1 activity is calculated by comparing kynurenine levels in treated vs. vehicle-treated wells (vehicle control defined as 0% inhibition). Cell viability is assessed by MTT assay (absorbance at 570 nm) to exclude cytotoxicity as the cause of reduced kynurenine production.
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| Animal Protocol |
Female BALB/c or C57BL/6 mice (6-8 weeks old, 18-22 g) are implanted subcutaneously with CT26 or B16 cells (1 × 10⁶ cells in 100 uL PBS/Matrigel). When tumors reach ~100 mm3 (7-10 days post-implantation), mice are randomized into treatment groups (n=8-10). (Rac)-IDO1-IN-5 is formulated in 5% DMSO + 30% PEG300 + 5% Tween 80 + 60% saline and administered orally at 10-100 mg/kg once or twice daily for 14-21 days. Tumor volumes are measured by calipers every 2-3 days using the formula V = (length × width2)/2. At study termination, plasma and tumors are collected for kynurenine and tryptophan analysis by LC-MS/MS. Tumor-infiltrating lymphocytes (TILs) are analyzed by flow cytometry (CD45+, CD3+, CD4+, CD8+, FoxP3+).
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| ADME/Pharmacokinetics |
(Rac)-IDO1-IN-5 is the racemate of the active S-enantiomer. The pharmacokinetic properties of the racemate are similar to those of the individual enantiomers in rodents. Following oral administration (10 mg/kg), the compound achieves Cmax of 1-3 uM within 1-2 hours, with a terminal half-life of 4-6 hours. Oral bioavailability is moderate (30-50%). The compound crosses the blood-brain barrier (BBB), achieving brain-to-plasma ratios of 0.3-0.5. Metabolism is primarily via CYP3A4-mediated oxidation.
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| Toxicity/Toxicokinetics |
Preclinical toxicity studies of (Rac)-IDO1-IN-5 in rodents show that a 14-day repeat-dose toxicity study (oral administration up to 300 mg/kg/day) is generally well tolerated. No mortality or significant body weight loss is observed at doses ≤ 100 mg/kg/day. At doses ≥ 200 mg/kg/day, mild gastrointestinal distress (loose stools, decreased food intake) and transient elevations in liver enzymes (ALT, AST) are noted. Histopathological examination reveals no target organ toxicity. No hERG inhibition (IC50 > 30 uM) or mutagenicity (Ames test negative).
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| References | |
| Additional Infomation |
IDO1 is a key enzyme in the tryptophan-kynurenine pathway that is often overexpressed in cancers, leading to immune suppression. (Rac)-IDO1-IN-5 was discovered by Eli Lilly and Company as part of their IDO1 inhibitor program. The compound‘s unique binding mode (interaction with apo-IDO1 rather than holo-IDO1) distinguishes it from other IDO1 inhibitors that target heme-bound IDO1. (Rac)-IDO1-IN-5 is a research tool for studying IDO1 biology, validating IDO1 as a cancer immunotherapy target, and developing combination strategies with checkpoint inhibitors. It is also valuable for comparing the pharmacological properties of the racemate versus its individual enantiomers.
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| Molecular Formula |
C23H25FN2O3
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|---|---|
| Molecular Weight |
396.454609632492
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| Exact Mass |
396.184
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| CAS # |
2166616-74-4
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| Related CAS # |
LY-3381916;2166616-75-5;(S)-LY-3381916;2166616-76-6
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| PubChem CID |
132209615
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| Appearance |
White to off-white solid powder
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| LogP |
2.9
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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 |
4
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| Heavy Atom Count |
29
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| Complexity |
587
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| Defined Atom Stereocenter Count |
0
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| SMILES |
FC1C=CC(=CC=1)C(NC(C)C1C=CC2=C(C=1)CCN2C(C1CCOCC1)=O)=O
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| InChi Key |
NUBWFWVVKLRSHS-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C23H25FN2O3/c1-15(25-22(27)16-2-5-20(24)6-3-16)18-4-7-21-19(14-18)8-11-26(21)23(28)17-9-12-29-13-10-17/h2-7,14-15,17H,8-13H2,1H3,(H,25,27)
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| Chemical Name |
4-fluoro-N-[1-[1-(oxane-4-carbonyl)-2,3-dihydroindol-5-yl]ethyl]benzamide
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| Synonyms |
(Rac)IDO1IN5 (Rac) IDO1 IN 5
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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 (~63.06 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.31 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to 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 (6.31 mM) (saturation unknown) 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. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.31 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5224 mL | 12.6119 mL | 25.2239 mL | |
| 5 mM | 0.5045 mL | 2.5224 mL | 5.0448 mL | |
| 10 mM | 0.2522 mL | 1.2612 mL | 2.5224 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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