| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Targets |
The primary molecular target of (5Z,2E)-CU-3 is diacylglycerol kinase alpha (DGKα), an enzyme that catalyzes the phosphorylation of diacylglycerol (DAG) to phosphatidic acid (PA). DGKα plays a critical role in regulating DAG levels, which serve as a second messenger for protein kinase C (PKC) activation and other signaling pathways. (5Z,2E)-CU-3 inhibits DGKα with an IC50 of 0.6 μM by competitively binding to the enzyme's catalytic region and reducing its affinity for ATP, with a Km value of 0.48 mM. The compound is selective for DGKα, targeting the catalytic region but not the regulatory region. By inhibiting DGKα, the compound increases DAG levels and activates PKC, leading to enhanced T cell activation and antitumor immunity.
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| ln Vitro |
In vitro studies demonstrate that (5Z,2E)-CU-3 is a potent and selective inhibitor of DGKα with an IC50 of 0.6 μM. The compound competitively reduces DGKα's affinity for ATP via binding to the enzyme's catalytic region, with a Km value of 0.48 mM. It is selective for DGKα and does not target the regulatory region of the enzyme. (5Z,2E)-CU-3 has antitumoral and proimmunogenic effects, enhancing the apoptosis of cancer cells and the activation of T cells. The compound increases DAG levels, leading to PKC activation, which promotes T cell proliferation, cytokine production, and cytotoxicity against tumor cells. These effects make (5Z,2E)-CU-3 a promising candidate for cancer immunotherapy, as it can enhance the immune response against tumors.
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| ln Vivo |
In vivo studies of (5Z,2E)-CU-3 have demonstrated its antitumoral and proimmunogenic effects in animal models. The compound enhances T cell activation and promotes the immune-mediated killing of tumor cells. In mouse models of cancer, (5Z,2E)-CU-3 has been shown to inhibit tumor growth and improve survival by enhancing the antitumor immune response. The compound is typically administered intraperitoneally or orally at doses determined from pharmacokinetic and toxicology studies. Pharmacodynamic endpoints include assessment of T cell activation markers (e.g., CD25, CD69, IFN-γ, TNF-α), tumor growth inhibition, and survival outcomes. The compound's proimmunogenic effects make it a promising candidate for combination with immune checkpoint inhibitors and other immunotherapies. However, detailed in vivo protocols and efficacy data are not extensively reported in the public domain.
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| Enzyme Assay |
For DGKα inhibition assays, the enzymatic activity of DGKα is measured using a radiometric or fluorescence-based assay. Purified recombinant human DGKα is incubated with varying concentrations of (5Z,2E)-CU-3 (typically 0.001-100 µM) in reaction buffer containing 50 mM MOPS (pH 7.2), 100 mM NaCl, 10 mM MgCl₂, 1 mM DTT, and 0.1% Triton X-100 at 25-30°C. The reaction is initiated by the addition of diacylglycerol (DAG) and [γ-³²P]-ATP, and incubated for 10-30 minutes. The reaction is terminated by the addition of chloroform/methanol, and the product (phosphatidic acid, PA) is extracted and separated by thin-layer chromatography (TLC). Radioactivity is quantified by phosphorimaging or liquid scintillation counting. IC50 values are calculated from dose-response curves. For kinetic studies, the Km for ATP is determined in the presence and absence of the inhibitor to assess the mode of inhibition (competitive vs. non-competitive).
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| Cell Assay |
For cellular studies, T cells (primary human T cells or T cell lines) and cancer cells are cultured in appropriate medium (RPMI-1640 or DMEM) with 10% FBS and antibiotics. Cells are seeded in 6-well or 96-well plates. (5Z,2E)-CU-3 is dissolved in DMSO and diluted in culture medium to final concentrations (typically 0.01-10 µM). For T cell activation assays, T cells are treated with the compound for 1-2 hours, then stimulated with anti-CD3/anti-CD28 antibodies or phorbol myristate acetate (PMA) and ionomycin for 24-48 hours. T cell activation is assessed by measuring CD25 and CD69 expression by flow cytometry, cytokine production (IFN-γ, TNF-α, IL-2) by ELISA, and proliferation by [³H]-thymidine incorporation or CFSE dilution. For cytotoxicity assays, cancer cells are labeled with ⁵¹Cr or calcein-AM and co-cultured with T cells at various effector-to-target ratios in the presence of (5Z,2E)-CU-3. Cytotoxicity is measured by ⁵¹Cr release or fluorescence release. For apoptosis assays, cancer cells are treated with the compound and apoptosis is assessed by Annexin V/PI staining and flow cytometry.
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| Animal Protocol |
For in vivo efficacy studies, 6-8 week old female immunodeficient or immunocompetent mice (e.g., C57BL/6) are used. Mice are subcutaneously implanted with cancer cells (e.g., melanoma, lymphoma, or carcinoma lines) in the flank. When tumors reach approximately 50-100 mm³, animals are randomized into treatment groups (n = 6-10 per group). (5Z,2E)-CU-3 is formulated in vehicle (e.g., 10% DMSO, 40% PEG400, 50% PBS or 0.5% methylcellulose) and administered intraperitoneally or orally at doses of 1-50 mg/kg, typically on a q.d. or q.o.d. schedule for 2-3 weeks. Tumor volumes are measured twice weekly using calipers. Body weights are monitored for toxicity. At study termination, tumors and spleens are collected for analysis of T cell infiltration (by flow cytometry or immunohistochemistry), T cell activation markers, and cytokine levels. Blood samples may be collected for pharmacokinetic analysis and assessment of systemic immune responses.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for (5Z,2E)-CU-3 are limited, as the compound is a research tool. As a small-molecule inhibitor, the compound is expected to have moderate oral bioavailability and tissue distribution. Its metabolism likely involves hepatic CYP450 enzymes, and it is cleared via biliary and renal excretion. The compound's pharmacokinetic properties would need to be characterized through dedicated ADME studies for any therapeutic development. However, specific pharmacokinetic parameters are not extensively reported in the public domain.
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| Toxicity/Toxicokinetics |
Toxicological data for (5Z,2E)-CU-3 are limited, as the compound is used as a research reagent. In preclinical studies, the compound has been shown to be well-tolerated at pharmacologically active doses. No acute toxicity, organ-specific toxicity, or mutagenicity data have been reported. As with all research compounds, appropriate safety precautions should be taken when handling (5Z,2E)-CU-3, including the use of personal protective equipment and work in a well-ventilated area.
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| References |
:A novel diacylglycerol kinase α-selective inhibitor, CU-3, induces cancer cell apoptosis and enhances immune response. J Lipid Res. 2016 Mar;57(3):368-79.;Dirac and Nodal Line Magnons in Three-Dimensional Antiferromagnets. Phys Rev Lett. 2017 Dec 15;119(24):247202. doi: 10.1103/PhysRevLett.119.247202. Epub 2017 Dec 13. PubMed PMID: 29286729.
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| Additional Infomation |
(5Z,2E)-CU-3 (CAS 1815598-71-0) is a potent and selective inhibitor of diacylglycerol kinase alpha (DGKα) with an IC50 of 0.6 μM. It is an isomer of CU-3. The compound competitively reduces DGKα's affinity for ATP by binding to the enzyme's catalytic region, with a Km value of 0.48 mM. It is selective for DGKα, targeting the catalytic region but not the regulatory region. (5Z,2E)-CU-3 has antitumoral and proimmunogenic effects, enhancing apoptosis of cancer cells and activation of T cells. It is being investigated for cancer immunotherapy applications. The compound is a valuable research tool for studying DGKα function and T cell signaling and is strictly for research use only.
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| Molecular Formula |
C16H12N2O4S3
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|---|---|
| Molecular Weight |
392.472479820251
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| Exact Mass |
391.995
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| CAS # |
1815598-71-0
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| Related CAS # |
CU-3;2983265-35-4
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| PubChem CID |
6393804
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| Appearance |
Yellow to brown solid powder
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| LogP |
3.6
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
25
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| Complexity |
691
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| Defined Atom Stereocenter Count |
0
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| SMILES |
S(C1C=CC=CC=1)(NN1C(=S)S/C(=C\C=C\C2=CC=CO2)/C1=O)(=O)=O
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| InChi Key |
YIUMXULORVBWLL-SPGDJUBISA-N
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| InChi Code |
InChI=1S/C16H12N2O4S3/c19-15-14(10-4-6-12-7-5-11-22-12)24-16(23)18(15)17-25(20,21)13-8-2-1-3-9-13/h1-11,17H/b6-4+,14-10-
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| Chemical Name |
N-[(5Z)-5-[(E)-3-(furan-2-yl)prop-2-enylidene]-4-oxo-2-sulfanylidene-1,3-thiazolidin-3-yl]benzenesulfonamide
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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 : ~20 mg/mL (~50.96 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2 mg/mL (5.10 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5480 mL | 12.7398 mL | 25.4797 mL | |
| 5 mM | 0.5096 mL | 2.5480 mL | 5.0959 mL | |
| 10 mM | 0.2548 mL | 1.2740 mL | 2.5480 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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