| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
NLRP3 inflammasome P2X7 Receptor; P2X7 receptor – irreversibly antagonizes P2X7 receptor activation by extracellular ATP; broad-spectrum P2 receptor inhibitor [1][2]
Oxidized ATP (oATP) targets P2 purinergic receptors, particularly the P2X7 receptor (P2X7R). It irreversibly antagonizes P2X7R activation by covalently modifying the receptor. oATP also inhibits P2X1, P2X2, P2X3, and P2X2/3 receptors, though with less selectivity. Additionally, it inhibits c-reactive protein (CRP)-induced NLRP3 inflammasome activation, linking P2X7 signaling to inflammatory pathways. |
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| ln Vitro |
In Vitro: OxATP (80 μM) directly inhibited T cell responses and suppressed T cell activation by altering dendritic cell (DC) function. Pre-exposing either responder T cells or APCs to oxATP (80 μM) significantly decreased Th17 responses; when both APCs and T cells were treated, the inhibition was significantly greater. OxATP did not significantly affect Th1 responses. [2]
OxATP (80 μM) neutralized the enhancing effect of ATP (100 μM) on Th17 response, but not on Th1 response, in vitro. [2] OxATP-treated bone marrow-derived dendritic cells (BMDCs) produced significantly smaller amounts of IL-23 and IL-6 compared to untreated BMDCs after LPS (100 ng/ml) stimulation, while IL-1 and IL-12 production was not significantly affected. OxATP-treated BMDCs were poorly stimulatory for IL-17⁺ IRBP-specific T cells compared to untreated BMDCs. [2] OxATP (80 μM) decreased the number of Foxp3⁺ cells among responder CD3⁺ T cells when exposed in culture. [2] OxATP (100 μM) was used as a broad-spectrum P2 receptor inhibitor in HUVECs, blocking CRP-induced NLRP3 inflammasome activation, caspase-1 activation, and IL-1β maturation. [1] In HUVEC, CRP (20 μg/mL, 24 hours)-induced caspase-1 activation and IL-1β maturation is inhibited by oxidized ATP trisodium salt (100 μM, 1 hour) [1]. In vitro, oxidized ATP trisodium salt blocks P2X7-mediated calcium flux and pore formation in immune cells such as macrophages and microglia. By irreversibly antagonizing P2X7R, it prevents ATP-induced NLRP3 inflammasome activation and subsequent IL-1beta and IL-18 release. oATP is also used to study the role of P2X receptors in cell death, cytokine secretion, and the pathogenesis of inflammatory and autoimmune diseases. |
| ln Vivo |
In Vivo: In a mouse experimental autoimmune uveitis (EAU) model (B6 mice immunized with IRBP₁₋₂₀/CFA), intraperitoneal injection of oxATP (300 μg/mouse, twice a week, starting 1 day post-immunization) almost completely abolished induced EAU as shown by fundoscopic and pathologic examination. Serum IL-17 was significantly decreased in oxATP-treated mice compared to controls, while IFN-γ was not significantly affected. [2]
In EAU mice, oxATP treatment significantly decreased the number of IL-17⁺ cells among in vivo primed responder T cells (15.2% in controls vs 8.1% in treated mice), while IFN-γ⁺ cells were minimally affected. Responder T cells from oxATP-treated mice produced significantly less IL-17 than T cells from non-treated mice. [2] IRBP-specific T cells isolated from oxATP-treated mice had significantly decreased ability to induce EAU upon adoptive transfer to naive mice. [2] Splenic DCs from oxATP-treated immunized mice produced significantly smaller amounts of IL-23 and IL-6 compared to untreated mice after LPS stimulation, while IL-1 and IL-12 production was not significantly different. [2] In HUVEC culture, oxATP (100 μM) blocked CRP-induced NF-κB activation, NLRP3 and pro-IL-1β expression, and NLRP3 inflammasome activation. [1] In B6 mice, oxidized ATP (300 μg/mouse, intraperitoneally, twice a week) and trisodium salt reduce experimental autoimmune uveitis (EAU)[2]. In vivo, oxidized ATP trisodium salt has been shown to ameliorate disease in mouse models of inflammation. For example, intraperitoneal administration of oATP (300 ug/mouse, twice weekly) effectively mitigates induced mouse experimental autoimmune uveitis (EAU). oATP also attenuates CRP-induced NLRP3 inflammasome activation in vivo, reducing LDL transcytosis across endothelial cells and thus having potential in atherosclerosis research. |
| Enzyme Assay |
Functional assays included measurement of cytokine production by ELISA, intracellular cytokine staining by flow cytometry, and assessment of T cell proliferation and differentiation. [1][2]
To evaluate oATP as a P2X7 antagonist, membrane preparations from P2X7-expressing cells (e.g., HEK293 or THP-1 cells) are incubated with radiolabeled ATP or a fluorescent P2X7 ligand (e.g., [3H]A-804598) in the presence or absence of oATP (0.1-100 uM). After a set incubation period at 4degC, bound ligand is separated by filtration, and radioactivity or fluorescence is measured to calculate inhibition of specific binding. |
| Cell Assay |
T cell preparation: αβ T cells were purified from spleen or draining lymph nodes of IRBP₁₋₂₀-immunized mice using an auto-MACS separator system. Purity was >95% by flow cytometry. Cells were cultured in RPMI 1640 with 10% fetal calf serum. [2]
Bone marrow dendritic cell (BMDC) generation: Bone marrow cells were cultured for 5 days in the presence of GM-CSF and IL-4 (10 ng/ml). BMDCs were generated and cultured with or without oxATP (80 μM). Cytokine levels (IL-1, IL-6, IL-12, IL-23) in culture medium were measured by ELISA. To determine antigen-presenting function, BMDCs were incubated with responder T cells under Th1- or Th17-polarizing conditions. [2] Th1 and Th17 response measurement: αβ T cells (1.8 × 10⁶) from immunized mice were co-cultured for 48 h with irradiated spleen cells (1.5 × 10⁶/well) as APCs and IRBP₁₋₂₀ (10 μg/ml) under Th1 (with IL-12, 10 ng/ml) or Th17 (with IL-23, 10 ng/ml) polarized conditions. Cytokine levels were measured by ELISA. The percentage of IFN-γ⁺ and IL-17⁺ T cells was determined by intracellular staining after 5 days. [2] HUVEC culture: Primary HUVECs were isolated from newborn umbilical cords and cultured in endothelial cell medium with 10% FBS, penicillin/streptomycin, and ECGS at 37°C with 5% CO₂. Cells were used between passages 2 and 9. [1] For cell-based assays, THP-1 monocytes or primary mouse macrophages are pre-incubated with oATP at concentrations ranging from 100-1000 uM for 30-60 minutes, then stimulated with ATP (1-5 mM) to activate P2X7 receptors. After stimulation for 1-4 hours, supernatants are collected to measure IL-1beta and IL-18 levels by ELISA. Pore formation can be assessed by uptake of ethidium bromide or YO-PRO-1 dye using a fluorescence plate reader. Cell viability is evaluated by LDH release assay. |
| Animal Protocol |
Animal/Disease Models: Induced mouse experimental autoimmune uveitis (EAU)[2]
Doses: 300 μg/mouse Route of Administration: ip Experimental Results: demonstrated almost undetected EAU, as shown by fundoscopic and pathologic examination. diminished serum IL-17 level. Mitigated the autoreactive T cell response. For in vivo studies, oATP is typically dissolved in sterile PBS or saline and administered intraperitoneally in mice at doses ranging from 100-500 ug per mouse (approximately 5-25 mg/kg). In the EAU model, mice are immunized with interphotoreceptor retinoid-binding protein (IRBP) on day 0, and oATP is injected i.p. twice weekly from day 3 onward. Ocular inflammation is evaluated by histopathological scoring and cytokine analysis. Other models include LPS-induced systemic inflammation or atherosclerosis-prone ApoE-/- mice. |
| ADME/Pharmacokinetics |
Pharmacokinetic data for oATP are limited. As a nucleotide analog, oATP is expected to be rapidly metabolized by ectonucleotidases in vivo, limiting its systemic half-life. It is typically formulated in PBS or saline for intraperitoneal administration. For optimal efficacy in animal models, repeated dosing (e.g., twice weekly) is required. Researchers should perform pilot PK studies to determine appropriate dosing intervals for their specific model.
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| Toxicity/Toxicokinetics |
The treatment regimen (300 μg/mouse, i.p., twice a week) was well tolerated in mice with no reported adverse effects. [2]
Toxicology data for oATP are limited. In cell-based assays, oATP can be used at concentrations up to 1 mM with manageable cytotoxicity. In animal studies, intraperitoneal administration of 300 ug/mouse twice weekly for several weeks is generally well tolerated with no reported severe adverse effects. However, comprehensive systemic toxicology studies have not been published. Care should be taken due to oATP's irreversible mechanism and potential for off-target effects. |
| References |
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| Additional Infomation |
Oxidized ATP (oxATP) is a small Schiff-base molecule that irreversibly antagonizes P2X7 receptor activation by extracellular ATP and is considered the most effective P2X7R inhibitor. It is a broad-spectrum P2 receptor inhibitor. OxATP has been shown to effectively block P2X7R activation and has therapeutic potential in autoimmune diseases. In the EAU model, oxATP treatment almost completely abolished induced disease, with the therapeutic effects involving functional changes in DCs, T cells, and regulatory T cells. OxATP not only directly inhibits T cell response but also suppresses T cell activation by altering the APC function of DCs. The inhibitory effect is stronger on Th17 than on Th1 response, possibly due to diminished Foxp3⁺ T cell activity offsetting the inhibitory effect on Th1 response. [2]
OxATP (100 μM) was used in HUVEC studies to block CRP-induced NLRP3 inflammasome activation, demonstrating its role as a broad-spectrum P2 receptor inhibitor. [1] oATP is a research tool and is not approved for clinical use. Its mechanism of action involves covalent modification of P2X7 receptors, leading to irreversible antagonism. This makes it useful for long-term blockade of P2X7 signaling in vitro and in vivo but also necessitates careful interpretation due to its broad-spectrum P2 receptor activity. oATP is commonly used as a positive control for P2X7 antagonism in pharmacological experiments. It is stored at -20degC, protected from moisture. |
| Molecular Formula |
C10H14N5O13P3.NA
|
|---|---|
| Molecular Weight |
528.15506
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| Exact Mass |
527.969
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| CAS # |
71997-40-5
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| PubChem CID |
57369844
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
17
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| Rotatable Bond Count |
12
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| Heavy Atom Count |
32
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| Complexity |
796
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1=NC(=C2C(=N1)N(C=N2)C(C=O)OC(COP(=O)(O)OP(=O)(O)OP(=O)(O)O)C=O)N.[Na]
|
| InChi Key |
KOCWSVPHHGJWHA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H14N5O13P3.Na/c11-9-8-10(13-4-12-9)15(5-14-8)7(2-17)26-6(1-16)3-25-30(21,22)28-31(23,24)27-29(18,19)20;/h1-2,4-7H,3H2,(H,21,22)(H,23,24)(H2,11,12,13)(H2,18,19,20);
|
| Chemical Name |
Adenosine 5'-triphosphate-2',3'-dialdehyde
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| Synonyms |
oATP; OxAT; KOCWSVPHHGJWHA-UHFFFAOYSA-N; CID 57369844; Adenosine 5'-triphosphate-2',3'-dialdehyde
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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, avoid exposure to moisture. |
| 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) |
H2O: ~50 mg/mL (87.6 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 | 1.8934 mL | 9.4668 mL | 18.9337 mL | |
| 5 mM | 0.3787 mL | 1.8934 mL | 3.7867 mL | |
| 10 mM | 0.1893 mL | 0.9467 mL | 1.8934 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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