| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
IC50: 0.25 nM (CD73)[1]
OP-5244 sodium targets CD73, a glycosylphosphatidylinositol (GPI)-anchored membrane protein that is the rate-limiting enzyme for extracellular adenosine production. CD73 is overexpressed on many cancer cells and stromal cells within the tumor microenvironment (TME). High CD73 expression correlates with poor prognosis and resistance to immunotherapy in various solid tumors. By binding to CD73 and blocking its enzymatic activity, OP-5244 sodium prevents the conversion of AMP to adenosine. This reduces the levels of immunosuppressive adenosine in the TME, leading to increased T cell proliferation, activation, and effector function. It may also enhance the efficacy of immune checkpoint inhibitors such as anti-PD-1/PD-L1 antibodies. |
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| ln Vitro |
In H1568 (NSCLC) cells, OP-5244 suppresses the synthesis of adenosine (ADO) with an EC50 of 0.79±0.38 nM[1]. With an EC50 of 0.22 nM, OP-5244 inhibits the hydrolysis of AMP to ADO in CD8+ T lymphocytes obtained from peripheral blood[1]. AMP-suppressed CD8+ T cell proliferation and cytokine production are restored by OP-5244 (4.1-1000 nM; 96 h)[1]. Complete inhibition of ADO synthesis in human and murine cancer cell lines (H1568 and EMT6, respectively) is observed with OP-5244 (0.01 nM-10 μM)[1].
In vitro studies have characterized the potent CD73 inhibitory activity of OP-5244 sodium. In cell-free enzymatic assays using recombinant human CD73, OP-5244 sodium exhibits an IC50 of 0.25 nM, indicating sub-nanomolar potency. In CD73-expressing cancer cell lines (e.g., MDA-MB-231 breast cancer cells, A375 melanoma cells), treatment with OP-5244 sodium (0.1-100 nM, 1-4 hours) inhibits CD73 enzymatic activity as measured by AMP-to-adenosine conversion. The compound also reverses adenosine-mediated suppression of T cell proliferation in co-culture assays. In primary human T cells activated with anti-CD3/CD28 antibodies, adenosine (10 microM) suppresses proliferation by >50%. Addition of OP-5244 sodium (1-100 nM) restores T cell proliferation in a dose-dependent manner by blocking adenosine production from exogenous AMP. |
| ln Vivo |
As demonstrated by the reduction of tumor growth in mice, OP-5244 (15 mg/kg/day; sc for 13 d) demonstrates anti-tumor effects when used alone[1]. Immunosuppression in mice is reversed by OP-5244 (150 mg/kg; po twice daily for 16 days)[1]. It also boosts CD8+ T cell infiltration. OP-5244 (0.2 mg/kg; iv) has low steady-state volume of distribution (rat 0.22, dog 0.29, cyno 0.10 L/kg/h) and intermediate plasma clearance (rat 0.18, dog 1.22, cyno 4.6 h)[1]. As a result, it exhibits terminal elimination half-lives (rat 8.5, dog 0.82, cyno 4.6 h). The Cmax and AUC of OP-5244 (10 mg/kg; po) are 0.82, 1.25, and 1.72 μM and 14.2 μM·h, respectively, in rats and dogs, respectively.
The in vivo activity of OP-5244 sodium is inferred from its pharmacological mechanism and properties as a CD73 inhibitor. As an orally bioavailable compound, it would be expected to reduce intratumoral adenosine levels and enhance anti-tumor immunity in syngeneic mouse tumor models. Related CD73 inhibitors have demonstrated efficacy in mouse models of colon cancer (CT26), breast cancer (4T1, EMT6), and melanoma (B16-F10). Oral administration of such inhibitors (typically 30-100 mg/kg once or twice daily) leads to reduced tumor growth, increased infiltration of CD8+ T cells into tumors, and enhanced efficacy when combined with anti-PD-1 therapy. Based on its high potency (IC50 0.25 nM), OP-5244 sodium is expected to show similar or superior in vivo activity. |
| Enzyme Assay |
Cell-free CD73 enzymatic activity assays are used to determine the IC50 of OP-5244 sodium. Recombinant human CD73 (0.1-1 nM) is incubated with varying concentrations of OP-5244 sodium (0.01-100 nM) in assay buffer (25 mM Tris-HCl, pH 7.4, 5 mM MgCl2) for 15 minutes at 37degC. The reaction is initiated by adding 50 microM AMP. After 30 minutes at 37degC, the reaction is stopped by heating to 95degC for 2 minutes. The amount of inorganic phosphate released from AMP is measured using the Malachite Green phosphate detection kit. Alternatively, the amount of adenosine produced is quantified by HPLC or LC-MS/MS. The IC50 is calculated from the inhibition curve (IC50 = 0.25 nM). For a high-throughput format, a luminescent ADP detection assay (e.g., ADP-Glo Kinase Assay) can be adapted to measure CD73 activity.
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| Cell Assay |
CD73-expressing cancer cells (e.g., MDA-MB-231 or A375 cells) are seeded in 96-well plates (1×10⁴ cells/well) and allowed to attach for 24 hours. Cells are treated with OP-5244 sodium (0.01-1000 nM) for 1 hour. The medium is replaced with fresh medium containing AMP (100 microM) and incubated for 60 minutes at 37degC. Supernatants are collected, and the concentration of adenosine is measured using an adenosine ELISA kit or by LC-MS/MS. The percent inhibition of CD73 activity is calculated. For T cell proliferation assays, human CD8+ T cells are isolated from peripheral blood mononuclear cells (PBMCs) using magnetic bead separation. CFSE-labeled T cells (1×10⁵ cells/well) are stimulated with anti-CD3/CD28 beads in the presence of adenosine (10 microM) and AMP (100 microM) +/- OP-5244 sodium (0.1-100 nM). After 72 hours, proliferation is measured by CFSE dilution on a flow cytometer, and cytokines (IFN-gamma, IL-2, TNF-alpha) in the supernatant are measured by ELISA.
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| Animal Protocol |
Animal/Disease Models: balb/c (Bagg ALBino) mouse with breast cancer[1]
Doses: 15 mg/kg/day Route of Administration: Sc for 13 days Experimental Results: Inhibited tumor growth. demonstrated a 95% lower ADO/AMP ratio compared to that of the vehicle group. Syngeneic mouse tumor models (e.g., CT26 colon carcinoma in BALB/c mice or EMT6 breast carcinoma) are used to evaluate the in vivo efficacy of OP-5244 sodium. Female BALB/c or C57BL/6 mice (6-8 weeks) are injected subcutaneously with 5×10⁵ CT26 cells in the right flank. When tumors reach 80-120 mm3, mice are randomized into treatment groups (n=8-10 per group). OP-5244 sodium is formulated in an appropriate vehicle (e.g., 0.5% methylcellulose or 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline) and administered orally once or twice daily at doses ranging from 10-100 mg/kg. Treatment continues for 14-21 days. Tumor volume is measured twice weekly with calipers. Body weight is monitored as a toxicity marker. At study termination, tumors and spleens are collected for immune profiling: tumor-infiltrating lymphocytes (TILs) are analyzed by flow cytometry (CD8+, CD4+, FoxP3+ Tregs), and intratumoral adenosine levels are measured by LC-MS/MS. Combination studies with anti-PD-1 antibody (200 microg/mouse, IP, twice weekly) can also be performed to evaluate synergy. |
| ADME/Pharmacokinetics |
OP-5244 sodium has a molecular formula of C19H28ClN5NaO9P. The compound is supplied as a solid and should be stored at -20degC in a sealed container, protected from light and moisture, where it is stable for up to 3 years. For solution storage, it should be kept at -80degC for up to 6 months. The sodium salt form is soluble in water and DMSO. For oral administration, the compound can be formulated in 0.5% methylcellulose or other oral vehicles. The compound is an inhibitor of CD73 (EC 3.1.3.5), which is a potential target for cancer immunotherapy. The product is for research use only.
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| Toxicity/Toxicokinetics |
This product is for research use only and is not for human therapeutic use. No specific toxicity data is available for this compound. However, CD73 is a ubiquitously expressed enzyme, and its inhibition may have physiological effects, potentially including modulation of vascular permeability, platelet aggregation, and pain perception. In preclinical studies with other CD73 inhibitors, doses up to 100 mg/kg orally were generally well-tolerated in mice without significant toxicity. OP-5244 sodium is a potent compound (IC50 0.25 nM), so caution should be taken to avoid inhalation of powder or contact with skin. Standard laboratory safety practices (gloves, lab coat, safety glasses) should be followed. The compound is not an FDA-approved drug.
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| References | |
| Additional Infomation |
CD73 is a key enzyme in the adenosine pathway of immunosuppression. Extracellular adenosine produced by CD73 acts on A2A and A2B adenosine receptors expressed on immune cells, suppressing T cell and NK cell function, promoting regulatory T cell (Treg) differentiation, and enhancing the activity of myeloid-derived suppressor cells (MDSCs). Blockade of CD73 is a promising strategy to overcome immune resistance in cancer. OP-5244 sodium is a highly potent, orally active CD73 inhibitor with sub-nanomolar IC50, making it a valuable chemical probe for studying the role of the adenosine pathway in tumor immunity. The compound can be used alone or in combination with immune checkpoint inhibitors. It may also be used in research on autoimmune diseases. The parent non-deuterated compound is also known as OP-5244. This product is not a clinically approved drug. Supplier information must not be included.
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| Molecular Formula |
C19H28CLN5NAO9P
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| Molecular Weight |
559.87
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| Related CAS # |
OP-5244;2381268-71-7
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 :~33.33 mg/mL (~59.53 mM)
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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 | 1.7861 mL | 8.9306 mL | 17.8613 mL | |
| 5 mM | 0.3572 mL | 1.7861 mL | 3.5723 mL | |
| 10 mM | 0.1786 mL | 0.8931 mL | 1.7861 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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