| 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 |
Ki: 41 nM (ENPP1)[1]
Enpp-1-IN-12 is a highly potent inhibitor of the ENPP1 enzyme, with a reported Ki of 41 nM. ENPP1 is the target, and its inhibition prevents the hydrolysis of extracellular nucleotides like ATP and cGAMP. This action is critical because ENPP1 hydrolyzes cGAMP, the natural agonist of the STING pathway, thereby dampening immune responses. |
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| ln Vitro |
In human and mouse hepatocytes, respectively, Enpp-1-IN-12 (5 μM) displays a half-life and intrinsic clearance of >120 min and <11.55 μL/min/million cells, and 61.88 min and 22.4 μL/min/million cells, respectively[1].
In vitro, Enpp-1-IN-12 shows potent biochemical inhibition with a Ki of 41 nM. In cell-based assays, it demonstrates good metabolic stability. In human and mouse hepatocytes, treatment with 5 microM of Enpp-1-IN-12 results in half-lives of >120 min and 61.88 min, and intrinsic clearance rates of <11.55 and 22.4 microL/min/million cells, respectively. |
| ln Vivo |
Lung cancer tumor growth in the LLC1 syngeneic murine tumor model is inhibited by Enpp-1-IN-12 (100 mg/kg; po)[1]. In healthy female BALB/c mice, Enpp-1-IN-12 (10 mg/kg; po) has a moderate oral bioavailability (F=45.1%), half-life (t1/2=1.04 h), and Cmax (303.10 ng/mL)[1]. In healthy female BALB/c mice, Enpp-1-IN-12 (1 mg/kg; iv) has a half-life of t1/2=0.76 h, a Cmax of 308.64 ng/mL, and a CL of 73.22 mL/min/kg[1].
In vivo, Enpp-1-IN-12 exhibits significant anti-tumor activity. In the LLC1 syngeneic murine tumor model, oral administration of Enpp-1-IN-12 at 100 mg/kg effectively inhibits lung cancer tumor growth. This anti-tumor effect is primarily due to the activation of the STING pathway and subsequent recruitment of immune cells into the tumor microenvironment. |
| Enzyme Assay |
The non-cellular protocol for Enpp-1-IN-12 is a standard ENPP1 enzyme inhibition assay. A typical procedure involves incubating recombinant human ENPP1 enzyme with the substrate bis(p-nitrophenyl)phosphate or a more sensitive fluorogenic substrate. Varying concentrations of the test compound are added, and after 30 minutes at 37degC, the reaction is stopped. The absorbance (for p-nitrophenol) or fluorescence is measured to calculate the Ki.
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| Cell Assay |
In vitro cellular assays for Enpp-1-IN-12 often focus on assessing its permeability and stability. To determine intrinsic clearance, hepatocytes are isolated from human and mouse livers and seeded. Enpp-1-IN-12 at 5 microM is added to the hepatocyte suspension. At various time points (0, 15, 30, 60, 90, and 120 min), samples are collected, and the reaction is stopped. The remaining compound concentration is measured by LC-MS/MS to calculate its half-life.
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| Animal Protocol |
For in vivo efficacy, a syngeneic mouse model is used. Female BALB/c mice are subcutaneously injected with LLC1 lung cancer cells. Once tumors are established, the mice are orally administered Enpp-1-IN-12, typically daily, at a dose of 100 mg/kg. Tumor volume is measured every 2-3 days with calipers. At the end of the study, tumors are excised and weighed, and immune cell infiltration is analyzed by flow cytometry.
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| ADME/Pharmacokinetics |
In healthy female BALB/c mice, oral administration of Enpp-1-IN-12 at 10 mg/kg shows moderate oral bioavailability (F=45.1%). It has a relatively short half-life (t1/2=1.04 h) and a Cmax of 303.10 ng/mL. After intravenous administration (1 mg/kg), the half-life is 0.76 h, Cmax is 308.64 ng/mL, and clearance (CL) is 73.22 mL/min/kg.
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| Toxicity/Toxicokinetics |
No specific toxicology data beyond the in vivo efficacy studies is provided. In the efficacy study at 100 mg/kg, no overt signs of toxicity were reported, but a standard toxicology panel would be required for development. The compound has been assessed for its effects on hepatocytes, but this was for stability, not toxicity.
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| References | |
| Additional Infomation |
Enpp-1-IN-12 is a key research tool for understanding the cGAS-STING pathway. The STING pathway is a critical mediator of the innate immune response to infection and cancer. By inhibiting ENPP1, the primary extracellular enzyme that degrades the STING agonist cGAMP, this compound provides a pharmacological method to potentiate immune surveillance against tumors and has significant potential for immunotherapy research.
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| Molecular Formula |
C16H18N6O3S
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|---|---|
| Molecular Weight |
374.417520999908
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| Exact Mass |
374.116
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| CAS # |
2631703-41-6
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| PubChem CID |
166565091
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| Appearance |
Off-white to gray solid powder
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| LogP |
2
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
26
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| Complexity |
487
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(NC1=CC=C(O)C(OC)=C1)(=O)C(SC1=C2C(=NC(N)=N1)NC=N2)CC
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| InChi Key |
ZDCMJGUVFRHQEH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H18N6O3S/c1-3-11(14(24)20-8-4-5-9(23)10(6-8)25-2)26-15-12-13(19-7-18-12)21-16(17)22-15/h4-7,11,23H,3H2,1-2H3,(H,20,24)(H3,17,18,19,21,22)
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| Chemical Name |
2-[(2-amino-7H-purin-6-yl)sulfanyl]-N-(4-hydroxy-3-methoxyphenyl)butanamide
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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: 50 mg/mL (133.54 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (2.67 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1 mg/mL (2.67 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 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6708 mL | 13.3540 mL | 26.7080 mL | |
| 5 mM | 0.5342 mL | 2.6708 mL | 5.3416 mL | |
| 10 mM | 0.2671 mL | 1.3354 mL | 2.6708 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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