| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
DCN1-UBC12[1]
DCN1 (defective in cullin neddylation 1, Ki = 12 nM) and DCN2 (Ki = 10.4 nM). DI-591 is a selective inhibitor of the DCN1-UBC12 interaction. It targets a specific step in the neddylation pathway, preventing the activation of the E3 ligase cullin 3 (CUL3). |
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| ln Vitro |
DI-591 (compound 44) has no discernible protein binding to DCN3, DCN4, or DCN5. Instead, it binds to DCN1 and DCN2 with Ki values of 12 nM and 10.4 nM, respectively. As a result, DI-591 is more than 1000 times more selective than recombinant human DCN3-5 proteins and shows very high binding affinity to recombinant human DCN1 and DCN2 proteins [1]. DI-591 (Compound 44; 0–10 μM; 1 hour; KYSE70 cells) binds to DCN1 and DCN2 proteins in cells and prevents DCN1 and UBC12 proteins from binding to each other [1]. The treatment of THLE2 cells with DI-591 (Compound 44; 10 μM) for 24 hours resulted in a significant increase in NQO1 and HO1 mRNA levels, which in turn caused the cells to upregulate HO1 protein. Interestingly, NRF2 mRNA levels are unaffected by DI-591 [1]. By specifically blocking cuLlin 3's neddylation, DI-591 causes an accumulation of NRF2 protein and the transcriptional activation of that protein. According to knockdown studies, cuLlin 3's neddylation is primarily regulated by DCN1, not DCN2, but not by other cuLlins. For the purpose of researching the function of cuLlin 3 CRL (CuLlin-RING E3 ubiquitin ligase) in biological processes and human diseases, DI-591 is a fantastic probe compound [1].
DI-591 binds to DCN1 and DCN2 with Ki values of 12 nM and 10.4 nM, respectively, and shows >1000-fold selectivity over DCN3-5. It selectively inhibits neddylation of CUL3 without affecting other cullins. In KYSE70 esophageal cancer cells, DI-591 (tested at 0-30 uM) disrupts DCN1-UBC12 association and elevates the mRNA expression of NQO1 and HO1, leading to increased HO1 protein expression. DI-591 is a chemical probe for studying the NRF2 pathway. |
| ln Vivo |
In a subcutaneous KYSE70 xenograft mouse model, DI-591 (0.6 mg/kg or 1.4 mg/kg, administered intraperitoneally, daily for 21 days) reduced tumor growth. Although it had no effect on the initial tumor growth rate, DI-591 decreased the final tumor size and weight, and suppressed tumor cell proliferation as measured by Ki-67 staining. It also significantly increased the expression of NRF2 protein in the tumors.
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| Enzyme Assay |
The activity of DCN1 is measured by its ability to promote the transfer of NEDD8 from the E2 enzyme UBC12 to cullin proteins. Recombinant DCN1 is incubated with UBC12 in the presence of NEDD8 and ATP. DI-591 is added at varying concentrations (1-1000 nM). The reaction is stopped, and the formation of the NEDD8-cullin conjugate is detected by Western blotting using an antibody specific for neddylated cullins. Quantification of the band intensity yields the IC50 or Ki values.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: KYSE70 cells Tested Concentrations: 0 μM, 1 μM, 3 Μm or 10 μM Incubation Duration: 1 hour Experimental Results: Potently bound to cellular DCN1 and DCN2 proteins. Enhanced the stability of DCN1 and DCN2 protein in a dose-dependent manner. RT-PCR[1] Cell Types: THLE2 cells Tested Concentrations: 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Robustly increases the mRNA levels of NQO1 and HO1. KYSE70 cells are treated with DI-591 (0-30 uM) for 24 hours. The cells are harvested, and the levels of neddylated CUL3, as well as total CUL3, are measured by Western blot to confirm target engagement. The levels of NRF2 and its target genes (HO1 and NQO1) are assessed by qPCR and Western blot. Cell proliferation is measured by the MTT assay. The effect of DI-591 on intracellular ROS levels can be measured using the fluorescent probe DCFH-DA. |
| Animal Protocol |
For in vivo efficacy studies, DI-591 is formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. It is administered intraperitoneally to mice bearing subcutaneous KYSE70 xenografts (0.6 mg/kg or 1.4 mg/kg, daily for 21 days). Tumor volume is measured with calipers. At the end of the study, tumors are harvested for Western blot analysis of neddylated cullins and NRF2 target proteins, and for immunohistochemistry to assess proliferation (Ki-67).
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| ADME/Pharmacokinetics |
DI-591 is cell-permeable. No detailed PK parameters (half-life, Cmax, AUC) are provided in the literature. It is typically dissolved in DMSO for in vitro studies. For in vivo administration, it is formulated in 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. The compound has a molecular weight of 585.80 g/mol and a molecular formula of C31H47N5O4S. It is stored as a solid at -20degC.
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| Toxicity/Toxicokinetics |
No detailed toxicological data are available. In the KYSE70 xenograft study at doses of 0.6 mg/kg and 1.4 mg/kg, no significant systemic toxicity or body weight loss was reported, suggesting a favorable safety profile at these doses. As a research chemical, standard safety precautions should be followed. It is not intended for human use.
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| References | |
| Additional Infomation |
DI-591 is a chemical probe for studying the NRF2 signaling pathway, which is activated in response to oxidative stress. By specifically inhibiting CUL3 neddylation, it stabilizes NRF2 and upregulates its target genes. This compound is a valuable tool for research into cancer, inflammation, and neurodegenerative diseases where the NRF2 pathway plays a protective role. It is not an FDA-approved drug.
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| Molecular Formula |
C31H47N5O4S
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|---|---|
| Molecular Weight |
585.800986528397
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| Exact Mass |
585.334
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| CAS # |
2245887-38-9
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| PubChem CID |
131750123
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| Appearance |
White to off-white solid powder
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| LogP |
4.3
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
41
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| Complexity |
848
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| Defined Atom Stereocenter Count |
2
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| SMILES |
S1C2C=C(C=CC=2N=C1C[C@H](C(N[C@@H](CNC(CCN1CCOCC1)=O)C1CCCCC1)=O)NC(CC)=O)C(C)C
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| InChi Key |
CNJKDQGPBAWNRY-IZZNHLLZSA-N
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| InChi Code |
InChI=1S/C31H47N5O4S/c1-4-28(37)33-25(19-30-34-24-11-10-23(21(2)3)18-27(24)41-30)31(39)35-26(22-8-6-5-7-9-22)20-32-29(38)12-13-36-14-16-40-17-15-36/h10-11,18,21-22,25-26H,4-9,12-17,19-20H2,1-3H3,(H,32,38)(H,33,37)(H,35,39)/t25-,26+/m0/s1
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| Chemical Name |
(2S)-N-[(1S)-1-cyclohexyl-2-(3-morpholin-4-ylpropanoylamino)ethyl]-2-(propanoylamino)-3-(6-propan-2-yl-1,3-benzothiazol-2-yl)propanamide
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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: 12.5 mg/mL (21.34 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (2.13 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 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL 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: ≥ 1.25 mg/mL (2.13 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 12.5 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. View More
Solubility in Formulation 3: ≥ 1.25 mg/mL (2.13 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7071 mL | 8.5353 mL | 17.0707 mL | |
| 5 mM | 0.3414 mL | 1.7071 mL | 3.4141 mL | |
| 10 mM | 0.1707 mL | 0.8535 mL | 1.7071 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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