| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| Other Sizes |
Purity: ≥98%
| Targets |
HBV(IC50= 53 nM )
HBV replication (IC50 = 53 nM in HepG2.2.15 cells) [1] HBV capsid protein (Cp) - no specific IC50/Ki reported; binding stoichiometry: at ≤1 drug per two dimers stabilizes capsids, at ≥1 drug per dimer destabilizes capsids [2] |
|---|---|
| ln Vitro |
In vitro, BAY 41-4109 has the capacity to misdirect and speed up capsid assembly. BAY 41-4109 stabilizes formed capsids up to a ratio of one inhibitor molecule for every two dimers [2]. In HepG2.2.15 cells, BAY 41-4109 exhibits an equivalent level of efficacy in suppressing HBV DNA release and cytoplasmic HBcAg levels, with IC50 values of 32.6 and 132 nM, respectively. The dose-dependent inhibition of HBV DNA and HBcAg suggests that the anti-HBV mechanisms are linked to and reliant on the rate of HBcAg inhibition[3].
BAY 41-4109 inhibited HBV replication in HepG2.2.15 cells with an IC50 of 53 nM and a CC50 (cytotoxicity against HepG2.2.15 cells) of 7 μM. [1] BAY 41-4109 enhanced the rate of HBV capsid assembly in vitro; as little as 0.2 μM drug (ratio of 1 drug molecule per 50 Cp149 dimers) increased assembly rate of 10 μM Cp149 in 150 mM NaCl at 40°C. [2] BAY 41-4109 misdirected assembly to form aberrant non-capsid polymers (balloon-shaped particles, spirals, sheets, tubes) at higher concentrations (≥5 μM BAY 41-4109 with 10 μM Cp149). [2] On preformed HBV capsids, BAY 41-4109 at a stoichiometry of ≤1 drug molecule per two dimers (e.g., 2 μM Cp with 1 μM drug) stabilized capsids without changing size; at ≥1 drug molecule per dimer (e.g., 2 μM Cp with 2 μM drug), capsids were destabilized and formed large non-capsid void polymers. This effect was temperature-dependent, with more void material observed at 4°C than at room temperature. [2] |
| ln Vivo |
With an efficacy similar to 3TC, BAY 41-4109 decreases viral DNA in the liver and plasma in a dose-dependent manner. Hepatitis B virus core antigen (HBcAg) in the livers of HBV-transgenic mice is decreased by BAY 41 -4109. Research on pharmacokinetics in mice has demonstrated quick absorption, a 30% bioavailability, and dose-proportional plasma concentrations in rats and dogs of roughly 60%[1]. By targeting the viral capsid, BAY41-4109 inhibits the production of viruses in vivo[2].
In HBV-transgenic mice (Tg[HBV1.3 fsX-3'5]), oral administration of BAY 41-4109 at doses of 3, 7.5, 15, and 30 mg/kg twice daily for 28 days reduced HBV-specific DNA in liver and plasma in a dose-dependent manner. At 15 and 30 mg/kg, significant reductions in liver HBV DNA (P<0.05) and plasma HBV DNA (P<0.05) were observed. Efficacy was comparable to or better than lamivudine (3TC) at 30 mg/kg. [1] Immunohistological analysis of livers from treated mice showed that BAY 41-4109 (30 mg/kg t.i.d. for 20 days) reduced cytoplasmic hepatitis B virus core antigen (HBcAg) to undetectable levels in both male and female mice, whereas 3TC had no effect on cytoplasmic HBcAg. Nuclear HBcAg was unaffected by BAY 41-4109. [1] |
| Enzyme Assay |
No classical enzyme assay described. For HBV capsid assembly studies: Assembly of truncated HBV capsid protein (Cp149) was monitored in real time by 90° light scattering at 400 nm in a fluorometer, with 3 nm band pass, at 40°C. Reactions were initiated by mixing Cp149 and drug with NaCl in 50 mM HEPES pH 7.5, 5 mM DTT. Light scattering was used to measure assembly kinetics. [2]
Size exclusion chromatography (SEC) on a Superose 6 10/300 column equilibrated with 50 mM HEPES pH 7.5, 50 mM NaCl was used to analyze assembly products. Elution volumes: void (6.0-7.2 ml), capsid (7.2-8.3 ml), dimer (15-16.5 ml). [2] Electron microscopy: Samples were visualized on a transmission electron microscope; images collected with CCD camera. [2] |
| Cell Assay |
MTT colorimetry is used to measure cellular metabolism. In 96-well plates, HepG2.2.15 cells are plated at a density of 2 × 103 cells per well. 20 μL of MTT solution (5 g/L) is added to each well after 8 days of treatment with varying concentrations of each antiviral compound, and the wells are then incubated for 4 hours at 37°C. To dissolve the crystals, 150 μL of DMSO is then added and stirred for 10 minutes. An ELISA reader is used to record absorbance values at 490 nm. The curve regression equation is used to compute the MTT values[3].
HepG2.2.15 cell assay: Cells were plated in 96-well plates and incubated with compound dilutions at 37°C, 5% CO2. Medium was changed at day 4. After 8 days, supernatants were collected, cells lysed, and HBV-DNA detected by dot-blot hybridization and quantified using a Lumilmager. [1] For HBV capsid assembly in vitro: Purified Cp149 dimers (truncated HBV capsid protein) were used. Assembly reactions were carried out in 50 mM HEPES pH 7.5, 150 mM or 500 mM NaCl, 5 mM DTT, at 40°C or room temperature, with various concentrations of BAY 41-4109. Products analyzed by SEC and EM. [2] |
| Animal Protocol |
Mice: For this study, HBV-transgenic mice are employed. The compounds (BAY 41-4109) are given to mice twice a day for 28 days in a suspension solution formulated with 0.5% tylose. The placebo is the 0.5% tylose. The animals are killed six hours after the last treatment, and the livers are taken out and frozen right away for further examination. The anesthetized animals' hearts are punctured to obtain blood[1].
HBV-transgenic mouse model (Tg[HBV1.3 fsX-3'5]): Male and female mice (~10 weeks old) with plasma HBV DNA levels of 10^7-10^8 genome equivalents/ml were used. BAY 41-4109 was formulated as a suspension in 0.5% Tylose (vehicle) and administered per os (oral gavage) at doses of 3, 7.5, 15, 30 mg/kg twice daily (b.i.d.) for 28 days. Six hours after the last treatment, animals were sacrificed; livers removed and frozen; blood collected by cardiac puncture. [1] For pharmacokinetic studies: Blood was collected after intravenous and oral administration at various time points (three animals per time point) up to 24 h post dose. Plasma was precipitated with acetonitrile and drug concentration determined by LC/MS/MS. [1] For capsid destabilization studies (in vitro): Preformed capsids (assembled from Cp149 dimers) were incubated with BAY 41-4109 at room temperature or 4°C overnight in 150 mM NaCl, then analyzed by SEC and EM. [2] |
| ADME/Pharmacokinetics |
In mice: clearance = 4.0 L/h/kg; intravenous half-life ≤1 h (text also states "short half-life of 2h" in mice); oral bioavailability = 31%; rapid absorption with tmax = 0.17 h. [1]
In rats: clearance = 1.75 L/h/kg; intravenous half-life = 2 h; oral bioavailability = 58%; tmax = 0.17 h. [1] In dogs: clearance = 0.72 L/h/kg; intravenous half-life = 5 h; oral bioavailability = 58%; tmax = 0.8 h. [1] Predicted human parameters: clearance = 0.46 L/h/kg; half-life = 4 h; bioavailability = 48% (allometric scaling). [1] Dose-proportional plasma concentrations observed. [1] |
| Toxicity/Toxicokinetics |
BAY-41-4109 (R-isomer) (CAS#: 298708-81-3) cytotoxicity: MTT assay in HepG2.2.15 cells showed TC50 = 19.3 μM. No signs of toxicity or reduced proliferative capacity were observed at concentrations used for antiviral assays (up to 1 μM). The therapeutic index (TI = TC50/EC50) is approximately 592 (19.3 μM / 32.6 nM). [3]
|
| References |
|
| Additional Infomation |
BAY 41-4109 is a pure R-enantiomer (the active enantiomer), whereas HAP-1 is a racemic mixture. [2]
Mechanism of action: BAY 41-4109 targets HBV capsid protein (Cp) and accelerates capsid assembly at substoichiometric ratios, acting as an allosteric activator. At higher concentrations, it misdirects assembly to form aberrant non-capsid polymers (sheets, tubes, spirals). It stabilizes preformed capsids at low ratios (≤1 drug per two dimers) but destabilizes them at higher ratios (≥1 drug per dimer), leading to void polymers. This suggests two functionally distinct classes of drug-binding sites on HBV capsids. [2] Unlike lamivudine (3TC), BAY 41-4109 reduces cytoplasmic HBcAg in livers of transgenic mice, indicating a different mode of action. [1] The compound is currently under investigation for treatment of HBV infection. [1] |
| Molecular Formula |
C18H13CLF3N3O2
|
|---|---|
| Molecular Weight |
395.762933492661
|
| Exact Mass |
395.065
|
| CAS # |
298708-81-3
|
| Related CAS # |
Bay 41-4109 racemate;298708-79-9;Bay 41-4109 (less active enantiomer);476617-51-3
|
| PubChem CID |
489221
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
3.454
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
27
|
| Complexity |
645
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
CC1=C([C@@H](N=C(N1)C2=C(C=C(C=N2)F)F)C3=C(C=C(C=C3)F)Cl)C(=O)OC
|
| InChi Key |
FVNJBPMQWSIGJK-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C18H13ClF3N3O2/c1-8-14(18(26)27-2)15(11-4-3-9(20)5-12(11)19)25-17(24-8)16-13(22)6-10(21)7-23-16/h3-7,15H,1-2H3,(H,24,25)
|
| Chemical Name |
Methyl 4-(2-chloro-4-fluorophenyl)-2-(3,5-difluoropyridin-2-yl)-6-methyl-1,4-dihydropyrimidine-5-carboxylate
|
| Synonyms |
BAY-414109; BAY414109; BAY 414109; BAY-41-4109; BAY41-4109; BAY 41-4109
|
| 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) |
DMSO : 79~100 mg/mL ( 199.61~252.68 mM )
Ethanol : ~11 mg/mL Water : ˂1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.32 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 25.0 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: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: ≥ 2.5 mg/mL (6.32 mM) (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5268 mL | 12.6339 mL | 25.2678 mL | |
| 5 mM | 0.5054 mL | 2.5268 mL | 5.0536 mL | |
| 10 mM | 0.2527 mL | 1.2634 mL | 2.5268 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA