| Size | Price | Stock | Qty |
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| 5mg |
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| 50mg |
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| Targets |
P2X7 Receptor
HEI3090 targets the P2X7 receptor (P2X7R), a trimeric ATP-gated cation channel expressed on immune cells, including dendritic cells, macrophages, and lymphocytes. Activation of P2X7R by HEI3090 triggers the assembly and activation of the NLRP3 inflammasome, leading to caspase-1-mediated cleavage of pro-IL-18 and pro-IL-1beta into their active forms. Mature IL-18 released from dendritic cells then acts on NK cells and CD4+ T cells, promoting the production of IFN-gamma, a key cytokine for anti-tumor immunity. |
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| ln Vitro |
In vitro, HEI3090 (1-100 uM) stimulates P2X7R-expressing dendritic cells, leading to IL-18 production (measured by ELISA). In co-culture systems, HEI3090-treated dendritic cells activate NK cells and CD4+ T cells to produce IFN-gamma. The effect is blocked by the P2X7R antagonist A438079 or by knocking down P2X7R expression, confirming target specificity. HEI3090 also induces IL-1beta release in macrophages. It has no direct cytotoxicity on tumor cells (IC₅0 > 50 uM). In human PBMCs, HEI3090 (10-50 uM) enhances IFN-gamma production and CD8+ T cell activation.
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| ln Vivo |
In vivo, HEI3090 triggers a sustained antitumor response in transplantable and oncogene-induced mouse models of lung cancer. In a mouse model of non-small cell lung cancer (NSCLC, e.g., Lewis lung carcinoma or KRAS-driven lung cancer), intraperitoneal administration of HEI3090 (10-30 mg/kg) significantly reduces tumor growth and prolongs survival. HEI3090 enhances the efficacy of alphaPD-1 (anti-PD-1) therapy when used in combination, leading to tumor regression and long-lasting immunity. In the tumor microenvironment, HEI3090 increases NK cell and CD8+ T cell infiltration and activation. The compound is also effective in models of melanoma and colon cancer. No significant toxicity is observed at therapeutic doses.
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| Enzyme Assay |
For P2X7R activation assays, HEK293 cells stably expressing human P2X7R are seeded in black-walled 96-well plates. Cells are loaded with a calcium indicator (Fluo-4-AM, 5 uM, 30 min). HEI3090 (0.1-100 uM) is added, and fluorescence (Ex/Em 494/506 nm) is measured immediately. The EC₅0 for calcium influx is calculated from dose-response curves. For IL-18 ELISA, human monocyte-derived dendritic cells (moDCs) are seeded in 96-well plates (1 × 10⁵ cells/well) and treated with HEI3090 (1-50 uM) for 24 h. IL-18 in the supernatant is measured by ELISA. The P2X7R antagonist A438079 (10 uM) is added 30 min before HEI3090 to block the response. For selectivity, the compound is tested against other P2X receptors (P2X1, P2X2, P2X3, P2X4) to confirm P2X7R specificity.
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| Cell Assay |
For cellular assays, human PBMCs (peripheral blood mononuclear cells) are isolated from whole blood by Ficoll-Paque density gradient centrifugation. PBMCs are seeded in 96-well plates (2 × 10⁵ cells/well) in RPMI-1640 with 10% FBS. HEI3090 (1-50 uM) is added for 24-48 h. IFN-gamma, IL-18, and IL-1beta in the supernatant are measured by ELISA. NK cell activation is assessed by flow cytometry using anti-CD69, anti-CD25, and anti-granzyme B antibodies. CD4+ and CD8+ T cell activation is assessed by IFN-gamma and granzyme B intracellular staining. For dendritic cell-T cell coculture, moDCs are treated with HEI3090 (10 uM) for 24 h, washed, and co-cultured with autologous CD4+ T cells or NK cells at a 1:10 ratio for 48-72 h. IFN-gamma is measured by ELISA, and T cell proliferation is measured by CFSE dilution. For cytotoxicity, HEK293 or J774 cells are treated with HEI3090 (1-200 uM) for 48 h; IC₅0 > 100 uM.
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| Animal Protocol |
For in vivo efficacy studies in a subcutaneous LLC (Lewis lung carcinoma) model, female C57BL/6 mice (6-8 weeks, n=10-12) are inoculated with 5 × 10⁵ LLC cells in 100 uL PBS. When tumors reach 100-150 mm3 (day 7-10), mice are randomized. HEI3090 is formulated in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline and administered intraperitoneally at 10-30 mg/kg once daily for 14 days. For combination therapy, anti-mouse PD-1 antibody (200 ug/mouse, IP, every 3 days) is co-administered. Tumor volumes are measured with calipers every 2-3 days. At study termination, tumors are harvested for flow cytometry (CD45+, CD3+, CD4+, CD8+, NK1.1+, FoxP3+, IFN-gamma+, granzyme B+), for immunohistochemistry (CD8, CD56, PD-1), and for cytokine analysis (IFN-gamma, IL-18, IL-1beta) by ELISA of tumor lysates. For survival studies, treatment continues for 28 days or until tumors reach 2000 mm3. For a KRAS-driven lung cancer model (e.g., Krasᴸˢᴸ-G¹2ᴰ/+; p53ˡˣ/ˡˣ mice), HEI3090 is administered by IP injection (10-30 mg/kg) 3×/week for 4-8 weeks. Lung tumor burden is assessed by micro-CT and H&E histology. Blood is collected for PK analysis. All IACUC guidelines apply.
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| ADME/Pharmacokinetics |
HEI3090 is soluble in DMSO (100 mg/mL). After IP administration in mice (10-30 mg/kg), Tmax is 0.5-1 h, and elimination half-life (t½) is 2-4 h. Plasma protein binding is moderate (60-80%). The compound is metabolized by CYP3A4 and excreted in feces and urine. Not orally bioavailable (<10%). Not for human use.
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| Toxicity/Toxicokinetics |
Preclinical toxicology: HEI3090 is well tolerated at doses up to 30 mg/kg IP in mice. No significant body weight loss, hepatotoxicity, or nephrotoxicity is reported. No genotoxicity. Immune-related adverse events (e.g., cytokine release syndrome) may occur at high doses (>100 mg/kg) but are not observed at therapeutic doses.
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| References | |
| Additional Infomation |
HEI3090 is a research compound in preclinical development as a P2X7R activator for cancer immunotherapy. It is not an FDA-approved drug. The compound can enhance the efficacy of alphaPD-1 therapy in non-small cell lung cancer and has potential for treating other solid tumors. For research use only.
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| Molecular Formula |
C18H15CL3N4O3
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| Molecular Weight |
441.70
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| CAS # |
2377167-56-9
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| Appearance |
White to off-white solid powder
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| SMILES |
N1(C(NC2=CC=C(Cl)N=C2)=O)C(=O)CC[C@H]1C(NCC1=CC=C(Cl)C=C1Cl)=O
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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) |
DMSO : ~50 mg/mL (~113.20 mM; with ultrasonication)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.66 mM) (saturation unknown) in 10% DMSO +90% Corn Oil (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 the 25.0 mg/mL clear DMSO stock solution to 900 μL corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2640 mL | 11.3199 mL | 22.6398 mL | |
| 5 mM | 0.4528 mL | 2.2640 mL | 4.5280 mL | |
| 10 mM | 0.2264 mL | 1.1320 mL | 2.2640 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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