Physicochemical Properties
| Molecular Formula | C27H25N5O2 |
| Molecular Weight | 451.52 |
| CAS # | 2922920-71-4 |
| Appearance | Typically exists as solid at room temperature |
| 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) |
Biological Activity
| ln Vitro |
In HEK293-A2AR cells, A2AR-antagonist-1 (0.001-10 μM; 30 minutes) lowers the levels of phosphorylated ERK generated by NECA [1]. In Jurkat T cells (human immortalized T line), A2AR-antagonist-1 (0.1-10 μM; 5 hours) enhances molecule expression while inhibiting NECA-induced immune molecule expression [1]. A2AR-antagonist-1 (0.1–10 μM; 48 h) improves the activation and activation of external T cells impact function and restores the cytotoxic function damage of OT-I mouse splenocytes (OT-I CTL) to MC38-OVA cells[1]. |
| ln Vivo | A2AR-antagonist-1 (100 mg/kg; wound cells; once daily for 23 days) showed remarkable anti-tumor effect in the C57BL/6 mouse model with breast cancer MC38 [1]. Metabokinetic analysis[1] Route dose (mg/kg) Cmax (ng/mL) AUC0-last (ng·h/mL) AUC0-t (ng·h/mL) t1/2 (h) F (%) iv 2 2584 5577 5565 0.93 / po 10 8823 24008 24003 2.35 86.1 |
| Cell Assay |
RT-PCR[1] Cell Types: Jurkat T cells Tested Concentrations: 0.1 μM, 1 μM and 10 μM Incubation Duration: 5 hrs (hours) Experimental Results: Reversal of NECA (1 μM)-induced upregulation of immunosuppressive molecules (LAG-3 and TIM-3) and NECA-induced down-regulation of effector molecules (GZMB, IFNG, and IL-2). Immunofluorescence[1] Cell Types: Cytotoxic T lymphocytes (OT-I CTL) from OT-I mouse splenocytes Tested Concentrations: 0.1 μM, 1 μM and 10 μM Incubation Duration: 48 hrs (hours) Experimental Results: OT-I CTL Relative lethality increases impression. |
| Animal Protocol |
Animal/Disease Models: Mouse MC38 xenograft model [1] Doses: 100 mg/kg Route of Administration: PO; one time/day for 23 days Experimental Results: Promote CD8+ T cells accumulation. Enhance anti-tumor immunity and promote tumor regression. There was no significant effect on mouse body weight. |
| References | [1]. Zhu C, et al. Discovery of Pyridinone Derivatives as Potent, Selective, and Orally Bioavailable Adenosine A2A Receptor Antagonists for Cancer Immunotherapy. J Med Chem. 2023 Mar 23. |
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 | 2.2147 mL | 11.0737 mL | 22.1474 mL | |
| 5 mM | 0.4429 mL | 2.2147 mL | 4.4295 mL | |
| 10 mM | 0.2215 mL | 1.1074 mL | 2.2147 mL |