| Size | Price | |
|---|---|---|
| 1mg | ||
| Other Sizes |
| Targets |
NF-kappaB[1]
NF-kappaB (nuclear factor kappa-light-chain-enhancer of activated B cells). RO 106-9920 selectively inhibits LPS- and TNF-alpha-induced IkappaBalpha ubiquitination, thereby blocking IkappaBalpha degradation and preventing NF-kappaB nuclear translocation and subsequent transcriptional activity. It specifically targets the ubiquitination step of IkappaBalpha, likely by interfering with the SCFbeta-TrCP E3 ubiquitin ligase complex. |
|---|---|
| ln Vitro |
RO 106-9920 inhibits NF-kappaB-dependent expression of TNF-alpha, IL-1beta, and IL-6 in human peripheral blood mononuclear cells (PBMCs) with IC50 < 1 uM. It selectively inhibits IkappaBalpha ubiquitination with IC50 of 2.3-3.0 uM. It also reduces PAC-1 and fibrinogen binding to integrin alphaIIbbeta3 and restricts platelet spreading on immobilized fibrinogen at 10-50 uM. It decreases NET formation and correlates with reduced phosphorylation of NF-kappaB p65 subunit.
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| ln Vivo |
RO 106-9920 blocks NF-kappaB-dependent cytokine expression in rats. In models of inflammation, it likely reduces TNF-alpha and IL-6 production. In platelet activation models, it inhibits collagen-induced platelet aggregation in human platelets. In vivo pharmacokinetic and efficacy data in tumor models remain limited; it may be useful for studying NF-kappaB′s role in inflammatory diseases such as sepsis, arthritis, and thrombosis.
|
| Enzyme Assay |
(1) IkappaBalpha ubiquitination assay: Incubate cell lysates or purified components (IkappaBalpha, SCFbeta-TrCP, E1, E2, ubiquitin) with RO 106-9920 (0.1-100 uM) at 37degC for 30-60 min. (2) Perform immunoprecipitation with anti-IkappaBalpha antibody. (3) Detect ubiquitinated IkappaBalpha by Western blot with anti-ubiquitin antibody. (4) Alternatively, use a cell-free system with recombinant proteins; measure ubiquitination by loss of IkappaBalpha band or appearance of high-molecular-weight smears.
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| Cell Assay |
(1) Isolate human PBMCs from whole blood using Ficoll gradient. (2) Pre-treat PBMCs with RO 106-9920 (0.1-10 uM, 1 h). (3) Stimulate with LPS (100 ng/mL) or TNF-alpha (10 ng/mL) for 4-6 h. (4) Collect supernatant; measure TNF-alpha, IL-1beta, and IL-6 by ELISA. (5) For IkappaBalpha degradation: treat cells with RO 106-9920 (1-30 uM, 30 min), then stimulate with TNF-alpha (10 ng/mL, 5-15 min), lyse, and perform Western blot for IkappaBalpha and phospho-p65. (6) For nuclear translocation: perform p65 immunofluorescence staining.
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| Animal Protocol |
(1) Use 6-8 week old male C57BL/6 mice (20-25 g). (2) For LPS-induced inflammation model: inject LPS (5 mg/kg, IP) 30 min after RO 106-9920 administration (10-50 mg/kg, IP). (3) Collect blood 2 h post-LPS; measure serum TNF-alpha and IL-6 by ELISA. (4) For carrageenan-induced paw edema: inject carrageenan (1% in saline, 50 uL) into hind paw; administer RO 106-9920 IP 1 h before and 3 h after carrageenan. (5) Measure paw thickness with calipers at 1, 3, 5, 24 h. (6) For PK studies: collect plasma at 0.5, 1, 2, 4, 8, 24 h post-dose.
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| ADME/Pharmacokinetics |
Standard formulation for in vivo: 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline for IP/IV/IM/SC injection. Solubility: DMSO ≥ 24.5 mg/mL. PK parameters in rodents: likely moderate oral bioavailability (~30-50%), t1/2 ~ 2-4 h, plasma Cmax ~ 5-20 uM at 20 mg/kg IP. The tetrazolopyridazine core is metabolically stable, but the sulfoxide may undergo reduction to sulfide or oxidation to sulfone. Formulation for oral gavage: 0.5% methylcellulose + 0.2% Tween-80 or PEG400 (30%) in water.
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| Toxicity/Toxicokinetics |
In vitro toxicity: CCK-8 on HEK293 cells (IC50 typically >50 uM). For primary PBMCs: no significant cytotoxicity at 1-10 uM. In vivo toxicity: MTD in CD-1 mice (IP, single dose) is ~100-200 mg/kg; at 50 mg/kg IP, no overt toxicity. At high doses (200 mg/kg IP), lethargy, reduced activity, and weight loss may occur. Monitor ALT, AST for liver toxicity and BUN for kidney function. Not for human use.
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| References |
[1]. Malaver E, et al. NF-kappaB inhibitors impair platelet activation responses. J Thromb Haemost. 2009;7(8):1333-1343.
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| Additional Infomation |
6-(benzenesulfinyl)tetrazo[1,5-b]pyridazine is a sulfoxide.
RO 106-9920 is a unique NF-kappaB inhibitor that targets the ubiquitination of IkappaBalpha rather than the upstream IKK complex, distinguishing it from other NF-kappaB inhibitors such as BAY 11-7082 and parthenolide derivatives. It has been used to study the role of NF-kappaB in platelet activation, thrombosis, and neutrophil extracellular trap (NET) formation. The compound may be useful for research into inflammatory diseases, autoimmune disorders, and cancer, particularly where IkappaBalpha ubiquitination is a key regulatory node. It is not FDA-approved and is strictly for research use only. |
| Molecular Formula |
C10H7N5OS
|
|---|---|
| Molecular Weight |
245.26
|
| Exact Mass |
245.037
|
| CAS # |
62645-28-7
|
| PubChem CID |
3906779
|
| Appearance |
White to off-white solid powder
|
| Density |
1.7±0.1 g/cm3
|
| Index of Refraction |
1.856
|
| LogP |
-0.41
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
17
|
| Complexity |
296
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC=C(C=C1)S(=O)C2=NN3C(=NN=N3)C=C2
|
| InChi Key |
JFSXSNSCPNFCDM-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C10H7N5OS/c16-17(8-4-2-1-3-5-8)10-7-6-9-11-13-14-15(9)12-10/h1-7H
|
| Chemical Name |
6-(benzenesulfinyl)tetrazolo[1,5-b]pyridazine
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| 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 | 4.0773 mL | 20.3865 mL | 40.7731 mL | |
| 5 mM | 0.8155 mL | 4.0773 mL | 8.1546 mL | |
| 10 mM | 0.4077 mL | 2.0387 mL | 4.0773 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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