| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg | |||
| Other Sizes |
| Targets |
IDO1 (Indoleamine 2,3-Dioxygenase 1). (S)-IDO1-IN-5 selectively binds to the heme-deficient apo-IDO1 rather than the mature heme-bound holo-IDO1, representing a unique mechanism of inhibition. By blocking IDO1 enzymatic activity, it prevents the conversion of tryptophan to kynurenine, thereby restoring tryptophan levels and reducing kynurenine pathway metabolites that mediate immune tolerance, T-cell anergy, and regulatory T-cell (Treg) differentiation.
|
|---|---|
| ln Vitro |
(S)-IDO1-IN-5 binds to IDOL (the target, likely IDO1) with an IC50 of less than 1.5 uM. It is a potent and selective inhibitor of IDO1 activity, with selectivity over IDO2 and TDO (tryptophan 2,3-dioxygenase). The S-enantiomer is significantly more active than the R-enantiomer or the racemic mixture in biochemical and cell-based assays.
|
| ln Vivo |
In mouse tumor models, LY-3381916 (the racemate) has been shown to reduce kynurenine levels in plasma and tumor tissues, enhance T-cell proliferation and function, and inhibit tumor growth when combined with checkpoint inhibitors (e.g., anti-PD-1). The S-enantiomer (S)-IDO1-IN-5 is expected to be the more potent form responsible for these in vivo pharmacological effects due to its higher binding affinity for apo-IDO1.
|
| Enzyme Assay |
Cell-free enzyme assays are performed using recombinant human IDO1 protein. The assay measures the conversion of L-tryptophan to N-formylkynurenine. IDO1 (10-50 nM) is pre-incubated with varying concentrations of (S)-IDO1-IN-5 (0.1 nM to 10 uM) for 15 minutes at 37degC in 50 mM potassium phosphate buffer (pH 6.5) containing 20 mM ascorbic acid, 20 uM methylene blue, and 0.5 mg/mL catalase. The reaction is initiated by adding L-tryptophan (100-500 uM) and incubated for 30-60 minutes at 37degC. The reaction is stopped with 30% trichloroacetic acid and further incubated at 65degC for 15 minutes to convert N-formylkynurenine to kynurenine. After centrifugation, the supernatant is mixed with Ehrlich‘s reagent (2% p-dimethylaminobenzaldehyde in acetic acid), and absorbance is measured at 480 nm. IC50 values are calculated from the inhibition of kynurenine formation.
|
| Cell Assay |
HeLa human cervical cancer cells, which constitutively express IDO1 upon IFN-gamma stimulation, are cultured in DMEM with 10% FBS. Cells are seeded in 96-well plates (30,000 cells/well) and treated with IFN-gamma (100 ng/mL) to induce IDO1 expression. Varying concentrations of (S)-IDO1-IN-5 (0.1 nM to 10 uM) are added, and cells are incubated for 24-48 hours. Kynurenine concentration in the culture supernatant is measured by the Ehrlich‘s reagent colorimetric assay or by HPLC-UV. The IC50 for inhibition of cellular IDO1 activity is calculated. Cell viability is assessed by MTT or CellTiter-Glo to ensure that any reduction in kynurenine is not due to cytotoxicity.
|
| Animal Protocol |
C57BL/6 mice bearing established syngeneic tumors (e.g., B16 melanoma or CT26 colon carcinoma) are treated with (S)-IDO1-IN-5 (10-100 mg/kg) or vehicle orally once or twice daily for 7-14 days. Plasma and tumor tissues are collected at various time points for measurement of tryptophan and kynurenine levels by LC-MS/MS. The kynurenine-to-tryptophan ratio (Kyn/Trp), an in vivo biomarker of IDO1 activity, is calculated to assess target engagement. Tumor growth is monitored, and immune cell infiltration (CD8+ T cells, Tregs) is analyzed by flow cytometry.
|
| ADME/Pharmacokinetics |
(S)-IDO1-IN-5 is a potent, selective, and brain-penetrant inhibitor of IDO1 (BBB permeable). It binds to heme-deficient apo-IDO1 rather than mature heme-bound IDO1, providing a unique selectivity mechanism. The compound is primarily metabolized by CYP450 enzymes in the liver, with moderate oral bioavailability and a half-life of 4-6 hours in rodents. It is highly protein bound (>90%) in plasma.
|
| Toxicity/Toxicokinetics |
Dose-limiting toxicity of IDO1 inhibitors in clinical trials includes fatigue, nausea, and diarrhea. (S)-IDO1-IN-5 is a research compound; preclinical toxicity data are limited to in vitro safety panels. In primary human hepatocytes, the compound has an IC50 > 50 uM for cytotoxicity, indicating a reasonable therapeutic window. No significant hERG inhibition has been observed at concentrations up to 10 uM.
|
| References | |
| Additional Infomation |
IDO1 is an attractive drug target for cancer immunotherapy because it suppresses anti-tumor immune responses. (S)-IDO1-IN-5 is the active S-isomer of LY-3381916, a compound discovered by Eli Lilly. The racemic mixture was evaluated in preclinical models of cancer, but clinical development has not been advanced. (S)-IDO1-IN-5 is a research tool for studying IDO1 biology, immune tolerance mechanisms, and the potential of IDO1 inhibition as a cancer immunotherapy strategy.
|
| Molecular Formula |
C23H25FN2O3
|
|---|---|
| Molecular Weight |
396.454609632492
|
| Exact Mass |
396.184
|
| CAS # |
2166616-76-6
|
| Related CAS # |
LY-3381916;2166616-75-5;(Rac)-LY-3381916;2166616-74-4
|
| PubChem CID |
132209650
|
| Appearance |
White to off-white solid powder
|
| LogP |
2.9
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
29
|
| Complexity |
587
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
FC1C=CC(=CC=1)C(N[C@@H](C)C1C=CC2=C(C=1)CCN2C(C1CCOCC1)=O)=O
|
| InChi Key |
NUBWFWVVKLRSHS-HNNXBMFYSA-N
|
| 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)/t15-/m0/s1
|
| Chemical Name |
4-fluoro-N-[(1S)-1-[1-(oxane-4-carbonyl)-2,3-dihydroindol-5-yl]ethyl]benzamide
|
| Synonyms |
(S)IDO1IN5 (S) IDO1 IN 5
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~300 mg/mL (~756.72 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 7.5 mg/mL (18.92 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 75.0 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: ≥ 7.5 mg/mL (18.92 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 75.0 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: ≥ 7.5 mg/mL (18.92 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
