| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
RNF5 (Ring Finger protein 5; also known as RMA1). RNF5 inhibitor inh-02 is a selective inhibitor of this E3 ubiquitin ligase. It blocks the ubiquitination activity of RNF5, preventing the degradation of its specific substrate proteins.
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| ln Vitro |
RNF5 inhibitor inh-02 is a potent inhibitor of the E3 ubiquitin ligase RNF5/RMA1. In bronchial epithelial cells homozygous for the F508del-CFTR mutation (the most common mutation in cystic fibrosis), this compound results in significant functional rescue of the CFTR chloride channel, with an EC50 of 2.2 uM. This rescue is achieved by inhibiting RNF5-mediated ubiquitination and degradation of the misfolded F508del-CFTR protein. It also regulates ATG4B and paxillin.
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| ln Vivo |
In vivo activity data for RNF5 inhibitor inh-02 are not detailed in the supplier literature. By rescuing functional CFTR expression, this compound has potential applications in research into cystic fibrosis, a disease caused by misfolded CFTR protein. Further in vivo studies would be required to assess its efficacy and safety in animal models of cystic fibrosis.
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| Enzyme Assay |
The activity of RNF5 can be measured in non-cellular assays using purified components of the ubiquitination cascade. Recombinant E1, E2 (e.g., UbcH5c), RNF5, and ubiquitin are incubated with a biotinylated substrate (e.g., a peptide or a purified protein) in the presence of ATP. RNF5 inhibitor inh-02 is added at varying concentrations (0.1-100 uM). The ubiquitination of the substrate is detected by Western blot or by a TR-FRET assay. A decrease in the ubiquitin signal indicates inhibition of RNF5 activity.
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| Cell Assay |
Cellular assays are performed using bronchial epithelial cells (e.g., CFBE41o- cells) that are homozygous for the F508del-CFTR mutation. Cells are treated with RNF5 inhibitor inh-02 (0.1-50 uM) for 24-48 hours. CFTR function is measured by a halide-sensitive fluorophore (e.g., SPQ) assay or by a YFP-based assay in a plate reader. Maturation of CFTR is assessed by Western blotting for band C (the mature, complex-glycosylated form) and band B (the immature form). The IC50 for functional rescue is calculated from the dose-response curve.
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| Animal Protocol |
For in vivo studies, RNF5 inhibitor inh-02 can be formulated in a vehicle such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline. It would be administered intraperitoneally or intratracheally to mouse models of cystic fibrosis (e.g., CFTR-/- mice or F508del humanized mice). Efficacy endpoints would include measurement of CFTR function in the nasal epithelium (by nasal potential difference) or in the lung, and assessment of lung histology, mucus accumulation, and inflammation.
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| ADME/Pharmacokinetics |
RNF5 inhibitor inh-02 has a molecular formula of C23H20N4S and a molecular weight of 384.50 g/mol. It is soluble in DMSO. The compound should be stored at 4degC, sealed, and away from moisture and light. For in vitro use, a 10 mM stock solution can be prepared in DMSO. Detailed pharmacokinetic parameters (half-life, oral bioavailability) are not reported in the supplier literature.
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| Toxicity/Toxicokinetics |
Detailed toxicological data for RNF5 inhibitor inh-02 are not available. It is intended for research use only and not for human consumption. Standard safety precautions for handling research chemicals should be followed, including the use of gloves and eye protection. The compound may be irritating to the skin, eyes, and respiratory tract.
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| References | |
| Additional Infomation |
RNF5 inhibitor inh-02 is a research-grade chemical, not an FDA-approved drug. It is used as a chemical probe to study the role of the ubiquitin-proteasome system in protein quality control. It has particular utility in validating RNF5 as a target for the treatment of cystic fibrosis, as inhibiting RNF5 allows for the rescue of the misfolded F508del-CFTR protein to the plasma membrane. It was described in a patent application (WO2012056087).
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| Molecular Formula |
C23H20N4S
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|---|---|
| Molecular Weight |
384.496703147888
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| Exact Mass |
384.14
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| CAS # |
324579-65-9
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| PubChem CID |
4151456
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| Appearance |
White to yellow solid powder
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| LogP |
5.1
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
28
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| Complexity |
594
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| Defined Atom Stereocenter Count |
0
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| SMILES |
S1/C(=N\C(\C2C=CC=CC=2)=N\C2C=CC=CC=2)/N(C(C)=N1)CC1C=CC=CC=1
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| InChi Key |
FYZFQLYZABRFAZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C23H20N4S/c1-18-26-28-23(27(18)17-19-11-5-2-6-12-19)25-22(20-13-7-3-8-14-20)24-21-15-9-4-10-16-21/h2-16H,17H2,1H3
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| Chemical Name |
N-(4-benzyl-3-methyl-1,2,4-thiadiazol-5-ylidene)-N'-phenylbenzenecarboximidamide
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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: 83.33 mg/mL (216.72 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 | 2.6008 mL | 13.0039 mL | 26.0078 mL | |
| 5 mM | 0.5202 mL | 2.6008 mL | 5.2016 mL | |
| 10 mM | 0.2601 mL | 1.3004 mL | 2.6008 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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