AB-423, a member of the sulfamoylbenzamide (SBA) class, is a potent HBV (hepatitis B virus) capsid assembly inhibitor that inhibits HBV replication with EC50/EC90 of 0.08-0.27 μM/0.33-1.32 μM in cells. AB-423 is currently under phase 1 clinical trials. AB-423 inhibited HBV genotypes A through D and nucleos(t)ide-resistant variants in vitro Treatment of HepDES19 cells with AB-423 resulted in capsid particles devoid of encapsidated pregenomic RNA and relaxed circular DNA (rcDNA), indicating that it is a class II capsid inhibitor. In a de novo infection model, AB-423 prevented the conversion of encapsidated rcDNA to covalently closed circular DNA, presumably by interfering with the capsid uncoating process. Molecular docking of AB-423 into crystal structures of heteroaryldihydropyrimidines and an SBA and biochemical studies suggest that AB-423 likely also binds to the dimer-dimer interface of core protein.
Physicochemical Properties
| Molecular Formula | C17H17F3N2O3S | |
| Molecular Weight | 386.39 | |
| Exact Mass | 386.09 | |
| Elemental Analysis | C, 52.84; H, 4.43; F, 14.75; N, 7.25; O, 12.42; S, 8.30 | |
| CAS # | 1572510-80-5 | |
| Related CAS # |
|
|
| PubChem CID | 90259477 | |
| Appearance | White to off-white solid powder | |
| LogP | 3.4 | |
| Hydrogen Bond Donor Count | 2 | |
| Hydrogen Bond Acceptor Count | 7 | |
| Rotatable Bond Count | 6 | |
| Heavy Atom Count | 26 | |
| Complexity | 584 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | CC[C@@H](C)NS(=O)(=O)C1=CC(=C(C=C1)F)C(=O)NC2=CC(=C(C=C2)F)F |
|
| InChi Key | BBLXLHYPDOMJMO-SNVBAGLBSA-N | |
| InChi Code | InChI=1S/C17H17F3N2O3S/c1-3-10(2)22-26(24,25)12-5-7-14(18)13(9-12)17(23)21-11-4-6-15(19)16(20)8-11/h4-10,22H,3H2,1-2H3,(H,21,23)/t10-/m1/s1 | |
| Chemical Name | (R)-5-(N-(sec-Butyl)sulfamoyl)-N-(3,4-difluorophenyl)-2-fluorobenzamide | |
| Synonyms | AB-423; AB 423; AB423 | |
| 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) |
Biological Activity
| Targets | HBV capsid |
| ln Vitro | Anti-HBV capsid assembly inhibitor AB-423. With EC50s of approximately 0.260 μM, AB-423 inhibits the production of rcDNA in AML12-HBV10 and HepDE19 cells. Additionally, AB-423 inhibits HBV DNA levels in culture supernatants of HepG 2.2.15 cells with an EC50 of 0.134 μM and suppresses cccDNA formation-dependent HBeAg production in the HepBHAe82 assay with an EC50 of 0.267 μM. Nonetheless, none of the three cell lines exhibit any cytotoxicity from AB-423[1]. |
| ln Vivo | In a mouse model of HBV, AB-423 (30 and 100 mg/kg, p.o. bid) inhibits HBV replication. In an HDI model of HBV in immunodeficient NOD-SCID mice, AB-423 (100 mg/kg, p.o. bid) combined with entecavir (ETV, 100 ng/mg, qd, p.o.) or 0.1 mg/kg dose of ARB-1467 potently inhibits serum HBV DNA[1]. |
| Cell Assay | HepBHAe82 (50,000 cells/well) are plated in 96-well tissue-culture treated microtiter plates in DMEM/F12 medium supplemented with 1% penicillin-streptomycin, 1% fetal bovine serum, and 1% tetracycline (1 μg/mL). The cells are then incubated for an entire night at 37°C and 5% CO2 in a humidified incubator. This is done to test the compound combinations. The cells are treated with inhibitors A and B the following day, with concentration ranges close to their individual EC50 values, after being transferred to a fresh medium.The inhibitors are diluted in either growth medium (ARB-1467 and ARB-1740) or 100% DMSO (ETV, TDF, and AB-423). The assay's final DMSO concentration is ≤0.5%. The effects of the two inhibitors on the inhibition of rcDNA production are assessed both individually and in combination. In the assay, the final DMSO concentration is 0.5%. The plates are kept in a humidified incubator with 5% CO2 and 37°C for nine days. After incubating for nine days, the medium is removed, and the cells undergo RNA extraction in order to quantify the precore mRNA level that is dependent on cccDNA[1]. |
| Animal Protocol |
Mice NOD receives 10 micrograms of the plasmid pHBV1.3 prior to the start of treatment.hydrodynamic injection (HDI; fast, high-volume injection into the tail vein; n = 6–8 animals per group) of CB17-Prkdcscid/J mice. Hepatitis B virus particles and other HBV products are produced when the 1.3-fold overlength copy of the HBV genotype D genome contained in this plasmid is expressed. Beginning on Day 0, AB-423 is given orally twice a day for seven days at a dose of 30 or 100 mg/kg. Beginning on Day 0, entecavir (ETV) at 100 ng/kg once daily for seven days in a row. On Day 0, a single intravenous bolus tail vein injection of ARB-1467 at a dose of 0.1 mg/kg is given. Days 0 (pre-dose), 4, and 7 are when blood is drawn for HBV biomarker analysis. Through the use of primer/probe sequences in a quantitative PCR assay, the amount of HBV DNA present in mouse serum is determined from the total extracted DNA[1]. |
| References |
[1]. Preclinical profile of AB-423, an inhibitor of Hepatitis B virus pgRNA encapsidation. Antimicrob Agents Chemother. 2018 May 25;62(6):e00082-18. |
Solubility Data
| Solubility (In Vitro) |
DMSO : 77~100 mg/mL ( 199.28~258.81 mM ) Ethanol : ~77 mg/mL Water : ˂1 mg/mL |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.47 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: ≥ 2.5 mg/mL (6.47 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 900 μL of corn oil and mix evenly. Solubility in Formulation 3: 10% DMSO+90% Corn Oil: ≥ 2.5 mg/mL (6.47 mM) (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5881 mL | 12.9403 mL | 25.8806 mL | |
| 5 mM | 0.5176 mL | 2.5881 mL | 5.1761 mL | |
| 10 mM | 0.2588 mL | 1.2940 mL | 2.5881 mL |