| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IC50: 20 nM (hARG1), 39 nM (hARG2), 39 nM (mARG1), 28 nM (rARG1)
Arginase 1 (ARG1) and Arginase 2 (ARG2), which are enzymes in the urea cycle that hydrolyze L-arginine to L-ornithine and urea. |
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| ln Vitro |
In cell-free enzymatic assays, OATD-02 hydrochloride has IC50 values of 20 nM for human ARG1, 39 nM for human ARG2, 39 nM for mouse ARG1, and 28 nM for rat ARG1, demonstrating its potent inhibitory activity across species.
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| ln Vivo |
In cancer cells, OATD-02 hydrochloride eliminates tumor immunosuppression induced by both arginases. In vitro, it has been shown to interfere with multiple immunosuppressive mechanisms driven by ARG1 and ARG2, thereby restoring effective antitumor immune responses.
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| Enzyme Assay |
A general protocol for evaluating ARG1 and ARG2 inhibition involves a colorimetric enzyme assay. Recombinant human ARG1 or ARG2 enzyme is incubated with various concentrations of the inhibitor (e.g., OATD-02 hydrochloride) in a reaction buffer (25 mM Tris-HCl, pH 7.5) at 37degC for 10 minutes. The substrate, L-arginine, is then added to a final concentration of 3 mM, and the reaction is further incubated at 37degC for 60 minutes. The reaction is terminated by the addition of a colorimetric detection reagent, which reacts with the produced urea to form a colored product. The absorbance is measured at 450 nm using a microplate reader. The IC50 value is calculated from the inhibition curve.
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| Cell Assay |
A general protocol for evaluating the cellular activity of arginase inhibitors is to measure L-arginine consumption and urea production in cancer cells or macrophages, which are known to upregulate arginase expression. Cells (e.g., melanoma or myeloid-derived suppressor cells) are seeded in 24-well plates and treated with various concentrations of the test compound (e.g., OATD-02 hydrochloride) for 24 to 72 hours. Cell culture supernatants are collected, and arginase activity is measured by quantifying the amount of urea produced using colorimetric assays. The effect on L-arginine levels can also be measured. A decrease in urea production and an increase in L-arginine levels indicate cellular arginase inhibition.
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| Animal Protocol |
A general protocol for evaluating the in vivo efficacy of OATD-02 hydrochloride involves a mouse xenograft model of melanoma. Immunocompromised or syngeneic mice are subcutaneously injected with B16-F10 melanoma cells. When the tumor volume reaches approximately 100-200 mm3, the mice are randomized into treatment and control groups. OATD-02 hydrochloride is administered orally once or twice daily at a dose of 10-100 mg/kg for 14-21 days. Tumor volume and body weight are measured every 2-3 days. At the end of the study, tumors are collected for Western blot analysis to assess arginase expression and for flow cytometry to analyze the tumor immune microenvironment (e.g., T-cell infiltration).
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| ADME/Pharmacokinetics |
A general protocol for evaluating the pharmacokinetic properties of OATD-02 involves administering a single dose orally (PO) and intravenously (IV) to male CD-1 mice. Blood samples are collected at various time points (e.g., 0.25, 0.5, 1, 2, 4, 8, and 24 hours) post-dose. Plasma concentrations of OATD-02 are quantified by a validated LC-MS/MS method. Pharmacokinetic parameters, including Cmax, Tmax, AUC, and oral bioavailability (F%), are calculated using non-compartmental analysis. OATD-02 is described as orally available, so this data would confirm its absorption and exposure profile.
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| Toxicity/Toxicokinetics |
General toxicity studies for OATD-02 would be performed in rodents. Sprague-Dawley rats would be administered OATD-02 hydrochloride orally at various dose levels (e.g., 50, 150, and 300 mg/kg/day) for 14 consecutive days. Clinical observations, body weight, and food consumption would be recorded daily. At the end of the study, blood samples would be collected for hematology and clinical chemistry analysis. Vital organs (liver, kidney, heart, etc.) would be harvested and weighed for histopathological examination. The no-observed-adverse-effect-level (NOAEL) would be determined based on the results.
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| References | |
| Additional Infomation |
OATD-02 is developed by OncoArendi Therapeutics. It is the first-in-class dual arginase inhibitor to enter human clinical trials. Phase I clinical trials have been initiated in patients with advanced solid tumors, including melanoma, to evaluate its safety, tolerability, pharmacokinetics, and preliminary antitumor activity in combination with immune checkpoint inhibitors.
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| Molecular Formula |
C12H26BCLN2O4
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|---|---|
| Molecular Weight |
308.61
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| Appearance |
White to off-white solid powder
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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) |
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
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2403 mL | 16.2017 mL | 32.4034 mL | |
| 5 mM | 0.6481 mL | 3.2403 mL | 6.4807 mL | |
| 10 mM | 0.3240 mL | 1.6202 mL | 3.2403 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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