| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| Other Sizes |
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| Targets |
NJH-2-056 targets the deubiquitinase OTUB1 (via covalent binding to C23 allosteric cysteine) and mutant F508del-CFTR protein (via lumacaftor moiety)[1]
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|---|---|
| ln Vitro |
NJH-2-056 engaged recombinant OTUB1 in gel-based activity-based protein profiling (ABPP) assays, confirming covalent binding to OTUB1. However, unlike its longer-linker analog NJH-2-057, it failed to stabilize F508del-CFTR protein levels in CFBE41o-4.7 human cystic fibrosis bronchial epithelial cells at tested concentrations (up to 10 μM), as measured by Western blotting[1]
The compound did not alter OTUB1 protein abundance in cells, indicating no off-target degradation effects[1] |
| Enzyme Assay |
OTUB1 Covalent Binding Assay: Recombinant OTUB1 was treated with NJH-2-056 and subsequently labeled with IA-rhodamine. Reduced fluorescence intensity (vs. DMSO control) confirmed covalent engagement at C23. LC-MS/MS of tryptic peptides verified selective modification at C23 without inhibiting catalytic activity[1]
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| Cell Assay |
F508del-CFTR Stabilization Assay: CFBE41o-4.7 cells expressing F508del-CFTR were treated with NJH-2-056 (concentrations up to 10 μM) for 24 hours. Cells were lysed, and protein extracts were analyzed by SDS-PAGE/Western blotting using CFTR-specific antibodies. GAPDH served as a loading control. Lumacaftor (VX-809) and OTUB1 recruiter EN523 were used as monotherapy controls[1]
OTUB1 Engagement Assay: Recombinant OTUB1 protein was pre-incubated with NJH-2-056 followed by IA-rhodamine probe labeling. Competition for labeling was visualized via in-gel fluorescence[1] |
| References | |
| Additional Infomation |
NJH-2-056 is a heterobifunctional DUBTAC (Deubiquitinase-Targeting Chimera) composed of the OTUB1 recruiter EN523 linked to lumacaftor (a CFTR corrector) via a C3 alkyl spacer. It served as a critical negative control, demonstrating that linker length (C3 alkyl) was insufficient to stabilize F508del-CFTR, unlike C5-C6 linkers (e.g., NJH-2-057)[1]
The compound’s design addresses cystic fibrosis pathology by recruiting OTUB1 to remove K48-linked polyubiquitin chains from misfolded F508del-CFTR, thereby rescuing it from proteasomal degradation[1] |
| Molecular Formula |
C41H40F2N6O8
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|---|---|
| Molecular Weight |
782.79
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| Exact Mass |
782.2875685
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| CAS # |
2858812-69-6
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| Appearance |
Off-white to pink solid powder
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| LogP |
4.4
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| SMILES |
CC1=C(N=C(C=C1)NC(=O)C2(CC2)C3=CC4=C(C=C3)OC(O4)(F)F)C5=CC(=CC=C5)C(=O)NCCCNC(=O)CCC6=CC=C(O6)N7CCN(CC7=O)C(=O)C=C
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| InChi Key |
SXPTYNZPHCBSCJ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C41H40F2N6O8/c1-3-34(51)48-20-21-49(35(52)24-48)36-15-11-29(55-36)10-14-33(50)44-18-5-19-45-38(53)27-7-4-6-26(22-27)37-25(2)8-13-32(46-37)47-39(54)40(16-17-40)28-9-12-30-31(23-28)57-41(42,43)56-30/h3-4,6-9,11-13,15,22-23H,1,5,10,14,16-21,24H2,2H3,(H,44,50)(H,45,53)(H,46,47,54)
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| Chemical Name |
3-[6-[[1-(2,2-difluoro-1,3-benzodioxol-5-yl)cyclopropanecarbonyl]amino]-3-methylpyridin-2-yl]-N-[3-[3-[5-(2-oxo-4-prop-2-enoylpiperazin-1-yl)furan-2-yl]propanoylamino]propyl]benzamide
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| Synonyms |
NJH-2-056; 2858812-69-6; SCHEMBL27164042; NJH2-056; SCHEMBL-27164042;
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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: ≥ 100 mg/mL (127.75 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 | 1.2775 mL | 6.3874 mL | 12.7748 mL | |
| 5 mM | 0.2555 mL | 1.2775 mL | 2.5550 mL | |
| 10 mM | 0.1277 mL | 0.6387 mL | 1.2775 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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