| Size | Price | Stock | Qty |
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| 25mg |
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| Targets |
Nexinhib20 specifically targets the protein-protein interaction (PPI) between the small GTPase **Rab27a** and its effector **JFC1** (synaptotagmin-like protein 1, Slp1). By blocking this interaction, it inhibits the exocytosis of azurophilic granules in neutrophils. It also inhibits Rac-1-GTP signaling. Unlike some earlier reports, recent studies confirm that Nexinhib20 does **not** inhibit Rac1 activation; its primary mode of action is the inhibition of Rab27a-JFC1 binding. Additionally, it has been reported to inhibit neutral sphingomyelinase 2 (nSMase2).
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| ln Vitro |
In vitro, Nexinhib20 inhibits the Rab27a-JFC1 interaction with an IC50 of 2.6 µM. It inhibits Rab27a with an IC50 of 330 nM. It effectively inhibits neutrophil exocytosis, adhesion, and β2 integrin activation. It inhibits myeloperoxidase (MPO) secretion induced by GM-CSF and fMLP in isolated human neutrophils with an IC50 of 0.33 µM. Nexinhib20 also inhibits the upregulation of adhesion molecules CD11b and CD66b, and suppresses extracellular production of superoxide anion. It prevents the expression of cytochrome b558 in the plasma membrane.
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| ln Vivo |
In vivo, Nexinhib20 displays anti-inflammatory activity. It reduces systemic inflammation mediated by neutrophils. Its therapeutic potential has been demonstrated in several preclinical models of inflammatory disease. Nexinhib20 is suitable for research applications focused on systemic inflammation and myocardial ischemia-reperfusion injury. It modulates vesicular trafficking and neutrophil activities in living organisms. Further details on specific in vivo dosing regimens and efficacy endpoints are available in the primary literature.
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| Enzyme Assay |
In vitro assays for Nexinhib20 typically involve measuring the inhibition of Rab27a-JFC1 binding using fluorescence polarization or other protein-protein interaction assays. The compound is dissolved in DMSO and diluted in assay buffer. IC50 values are calculated from dose-response curves. For neutrophil function assays, human neutrophils are isolated from peripheral blood and treated with Nexinhib20 at various concentrations (0.01-10 µM). Neutrophil exocytosis is stimulated with fMLP or GM-CSF, and released markers (e.g., MPO, lactoferrin) are measured by ELISA or enzymatic assays. Superoxide anion production and CD11b upregulation are assessed by standard methods.
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| Cell Assay |
For cell-based assays, primary human neutrophils or neutrophil-like cell lines (e.g., HL-60 differentiated cells) are cultured in RPMI-1640 with appropriate supplements. Cells are seeded in multi-well plates and pre-incubated with Nexinhib20 at various concentrations (typically 0.1-10 µM) for 15-30 minutes. Cells are then stimulated with agonists such as fMLP, PMA, or GM-CSF. Exocytosis is quantified by measuring granule marker release (e.g., MPO for azurophilic granules, lactoferrin for specific granules) in the supernatant. β2 integrin activation and adhesion are assessed by flow cytometry or adhesion assays. Cell viability is confirmed by standard assays. All treatments include vehicle controls.
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| Animal Protocol |
In vivo, Nexinhib20 is typically administered to rodents via intraperitoneal (IP) or intravenous (IV) injection. The compound is formulated in a suitable vehicle such as DMSO, PEG, or saline-based solutions. For inflammatory disease models (e.g., acute lung injury, peritonitis, or ischemia-reperfusion injury), Nexinhib20 is dosed at various regimens (e.g., 1-10 mg/kg). Endpoints include assessment of neutrophil infiltration (e.g., MPO activity in tissues), inflammatory cytokine levels, and histological evaluation of tissue damage. Blood and tissue samples are collected for pharmacokinetic and pharmacodynamic analysis. All procedures follow institutional animal care guidelines.
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| ADME/Pharmacokinetics |
Nexinhib20 (MW 300.31, C15H16N4O3) is soluble in DMSO (up to 100 mM or 30 mg/mL). It is a small, lipophilic molecule with good cell permeability. Detailed pharmacokinetic parameters (bioavailability, half-life, clearance) are not extensively reported in publicly available sources but are described in preclinical studies. The compound is typically stored at +4°C. For in vivo use, it is formulated in suitable vehicles to ensure adequate exposure. Further ADME studies are available in the primary literature.
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| Toxicity/Toxicokinetics |
Toxicology data for Nexinhib20 are limited in publicly available sources. The compound has been evaluated in preclinical models of inflammation and appears to be well-tolerated at efficacious doses. It selectively inhibits neutrophil exocytosis without affecting phagocytosis or NET production, suggesting a favorable safety profile regarding innate immune function. However, standard safety pharmacology and toxicology studies would be required for therapeutic development. The compound is for research use only and not intended for human therapeutic applications. Standard laboratory safety precautions should be followed.
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| References |
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| Additional Infomation |
Nexinhib20 is a research-grade chemical probe for studying neutrophil exocytosis, Rab27a-JFC1 interaction, and inflammatory diseases. Its primary applications include immunology, inflammation research, and drug discovery. The compound is not approved for clinical use and has not entered clinical trials. It is commercially available from various chemical suppliers for research purposes only. Its unique mechanism of specifically inhibiting neutrophil exocytosis while preserving other immune functions makes it a valuable tool for investigating the role of neutrophils in inflammatory diseases.
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| Molecular Formula |
C15H16N4O3
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|---|---|
| Molecular Weight |
300.312542915344
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| Exact Mass |
300.122
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| CAS # |
331949-35-0
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| PubChem CID |
5715188
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| Appearance |
White to yellow solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
508.2±60.0 °C at 760 mmHg
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| Flash Point |
261.2±32.9 °C
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| Vapour Pressure |
0.0±1.3 mmHg at 25°C
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| Index of Refraction |
1.601
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| LogP |
2.5
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
22
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| Complexity |
464
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(C)(C)C(=O)/C(=C/C1=CC(=CC=C1)[N+](=O)[O-])/N2C=NC=N2
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| InChi Key |
VKSOIYNNOFGGES-JYRVWZFOSA-N
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| InChi Code |
InChI=1S/C15H16N4O3/c1-15(2,3)14(20)13(18-10-16-9-17-18)8-11-5-4-6-12(7-11)19(21)22/h4-10H,1-3H3/b13-8-
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| Chemical Name |
4,4-Dimethyl-1-(3-nitrophenyl)-2-(1H-1,2,4-triazol-1-yl)-1-penten-3-one
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| Synonyms |
Nexinhib-20Nexinhib 20Nexinhib20
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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) |
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
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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 | 3.3299 mL | 16.6495 mL | 33.2989 mL | |
| 5 mM | 0.6660 mL | 3.3299 mL | 6.6598 mL | |
| 10 mM | 0.3330 mL | 1.6649 mL | 3.3299 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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