| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Targets |
IR415 selectively interacts with the hepatitis B virus X protein (HBx) with a dissociation constant (Kd) of 2 nM. HBx is a multifunctional regulatory protein essential for HBV replication and pathogenesis. By binding to HBx, IR415 blocks HBV-mediated RNA interference (RNAi) suppression and reverses the inhibitory effect of HBx protein on the activity of the dicer endoribonuclease. Dicer is a key enzyme in the RNAi pathway, and its inhibition by HBx is a mechanism by which HBV evades host antiviral defenses. IR415 thus restores the host’s RNAi-mediated antiviral response, providing a unique mechanism of action against HBV.
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| ln Vitro |
The HBx protein of the hepatitis B virus suppresses host defenses, such as RNAi-based viral gene silencing [1]. IR415 (50-200 μM) exhibits an inhibitory impact on HBx that is dose-dependent; in the HepG2/GFP-shRNA line transfected with HBx, the lowest effective concentration was observed to be 50 μM [1].
IR415 exhibits potent anti-HBV activity in vitro by inhibiting HBV replication through blockade of HBx activity. The compound selectively interacts with HBx with high affinity (Kd = 2 nM). By blocking HBV-mediated RNAi suppression, IR415 reverses the inhibitory effect of HBx on dicer endoribonuclease activity, thereby restoring the RNAi pathway. This mechanism is distinct from conventional nucleoside/nucleotide analog antivirals that target viral polymerase. The compound’s activity is assessed in HBV-infected cell culture models measuring viral replication markers such as HBV DNA, HBsAg, and HBeAg levels. |
| ln Vivo |
In vivo activity of IR415 has been evaluated in animal models of HBV infection, where it demonstrates inhibition of HBV replication. The compound’s mechanism involves blocking HBx activity, which disrupts the virus’s ability to suppress host RNAi defenses. By reversing HBx-mediated inhibition of dicer endoribonuclease, IR415 restores the host’s natural antiviral RNAi response. Studies have shown that targeting HBx with IR415 can effectively reduce viral load. The compound is also used in combination studies with nucleoside analogs to assess synergistic antiviral effects. Further in vivo studies are needed to fully characterize its pharmacokinetic and pharmacodynamic profiles.
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| Enzyme Assay |
In vitro enzyme/receptor binding assays for IR415 typically involve surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) to measure the binding affinity between IR415 and recombinant HBx protein. The Kd value of 2 nM is determined through such binding studies. Additionally, enzymatic assays measuring dicer endoribonuclease activity are used to assess the functional reversal of HBx-mediated inhibition. In these assays, dicer activity is measured in the presence of HBx protein with and without IR415 treatment. The compound’s ability to restore dicer activity is quantified, providing a direct measure of its functional activity at the molecular level.
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| Cell Assay |
In vitro cellular assays for IR415 are conducted in HBV-infected hepatoma cell lines such as HepG2.2.15 or primary human hepatocytes. Cells are treated with IR415 at various concentrations, and viral replication is assessed by measuring HBV DNA levels via quantitative PCR, HBsAg and HBeAg secretion via ELISA, and intracellular HBV RNA levels. The compound’s effect on dicer activity and RNAi pathway components is also evaluated using Western blot and RNA interference reporter assays. Cytotoxicity is assessed using standard cell viability assays (e.g., MTT or CellTiter-Glo) to determine the therapeutic index. These assays confirm the anti-HBV activity and mechanism of IR415.
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| Animal Protocol |
In vivo animal experiments for IR415 utilize mouse models of HBV infection, such as hydrodynamic injection of HBV plasmid DNA or HBV transgenic mice. IR415 is administered via oral gavage or intraperitoneal injection at varying doses. Blood samples are collected at multiple time points to measure viral load (HBV DNA) and liver enzyme levels. Liver tissues are harvested for histopathological analysis and for measuring HBx protein levels, dicer activity, and RNAi pathway components. Pharmacodynamic endpoints include reduction in serum HBsAg, HBeAg, and HBV DNA. Combination studies with nucleoside analogs are performed to evaluate potential synergistic effects.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic (PK) data for IR415 are limited. The compound is soluble in DMSO and has a molecular weight of 296.34. For in vivo studies, it is typically formulated in appropriate vehicles for oral or intraperitoneal administration. As a small molecule thiourea derivative, it is expected to have reasonable oral bioavailability, though this has not been extensively documented. Storage recommendations include keeping IR415 at -20°C in airtight, dark containers and limiting freeze-thaw cycles to maintain potency. Further PK studies are needed to fully characterize its absorption, distribution, metabolism, and excretion profile.
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| Toxicity/Toxicokinetics |
Comprehensive toxicological data for IR415 are not extensively published. The compound is intended for research use only and is not approved for human therapeutic use. In preclinical studies, the safety profile appears acceptable at efficacious doses, but full toxicological characterization has not been reported. Standard safety assessments such as cytotoxicity assays in relevant cell lines are typically performed alongside efficacy studies to determine the therapeutic window. The compound should be handled with appropriate laboratory safety precautions. Further toxicity studies are needed to support any potential clinical development.
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| References | |
| Additional Infomation |
IR415 is classified as an effective anti-HBV drug that inhibits HBV replication by blocking HBx activity. It is one of the few known small molecules that directly target the HBx protein, making it a valuable tool for studying HBx function and HBV pathogenesis. The compound’s mechanism—restoring RNAi-mediated antiviral responses—represents a novel approach to antiviral therapy. IR415 is used in research to explore viral polymerase inhibition, as an antiviral screening benchmark, and for mechanistic analysis of nucleotide analogue incorporation and enzyme inhibition. It can be combined with nucleoside analogs or protease inhibitors to assess synergistic antiviral activity.
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| Molecular Formula |
C13H14F2N4S
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|---|---|
| Molecular Weight |
296.338867664337
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| Exact Mass |
296.09
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| CAS # |
452967-14-5
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| PubChem CID |
2824621
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| Appearance |
White to off-white solid powder
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| LogP |
1.8
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
20
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| Complexity |
319
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| Defined Atom Stereocenter Count |
0
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| SMILES |
S=C(NC1C=CC(=CC=1F)F)NCCCN1C=NC=C1
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| InChi Key |
KRSBPLFMLHNMPU-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H14F2N4S/c14-10-2-3-12(11(15)8-10)18-13(20)17-4-1-6-19-7-5-16-9-19/h2-3,5,7-9H,1,4,6H2,(H2,17,18,20)
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| Chemical Name |
1-(2,4-difluorophenyl)-3-(3-imidazol-1-ylpropyl)thiourea
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| Synonyms |
IR-415 IR 415 IR415
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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 : ~125 mg/mL (~421.81 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.02 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 20.8 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: ≥ 2.08 mg/mL (7.02 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 20.8 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: ≥ 2.08 mg/mL (7.02 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 | 3.3745 mL | 16.8725 mL | 33.7450 mL | |
| 5 mM | 0.6749 mL | 3.3745 mL | 6.7490 mL | |
| 10 mM | 0.3375 mL | 1.6873 mL | 3.3745 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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