Hepatitis B Virus (HBV) Replication: AT-130 is a non-nucleoside analogue inhibitor of HBV replication. Its mechanism of action appears to be independent of interference with the RNA- or DNA-dependent activities of the HBV polymerase. Previous studies suggest it may inhibit the packaging of pregenomic viral RNA. No direct IC50/Ki values for specific enzyme targets are provided. [1]

AT-130 suppresses rtL180M (IC50=9.8 μM), rtM204I (IC50=35.6 μM), and Wt (IC50=2.4 μM) HBV [1]. HepG2 cells transduced with HBV baculovirus showed dose-dependent suppression of wt HBV replication when treated with AT-130 (0.1, 1, 5, 10, 100 μM) for 7 days. When AT-130 is present, encapsidated HBV DNA is reduced by 50% (IC50) at 2.5 μM and by 90% (IC90) at 18.5 μM [1]. At concentrations as high as 250 μM, AT-130 does not cause harm to HepG2 or Huh-7 cells [1]. By directly inhibiting the HBV endogenous DNA polymerase reaction in Huh 7 or HepG2 cells, AT-130 (0.005, 0.05, 0.5, 5, 50 μM) does not inhibit HBV DNA synthesis. While AT-130 has no effect on viral DNA polymerase activity or core protein translation, it suppresses the replication of HBV DNA in liver cancer cells [3]. While it decreases encapsidated RNA, AT-130 (2.5, 18.5 μM) had little effect on total HBV RNA. The activity of protein expression vectors and the synthesis of nucleocapsids or core proteins are unaffected by AT-130 [3].

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Inhibition of Wild-Type HBV Replication: In HepG2 cells transduced with recombinant baculovirus encoding wild-type HBV (genotype A, subtype adw2), AT-130 inhibited replication with IC50 = 2.40 ± 0.92 μM and IC90 = 18.5 μM, based on Southern blot analysis of intracellular replicative intermediates after 7 days of drug exposure. For comparison, lamivudine had IC50 = 0.064 ± 0.020 μM against wild-type HBV. [1]
Activity Against Lamivudine-Resistant HBV Mutants: AT-130 was tested against three nucleoside analogue-resistant HBV mutants: rtL180M, rtM204I, and rtL180M + rtM204V. The IC50 values were: 1.6 ± 0.4 μM for rtL180M; 4.2 ± 2.4 μM for rtM204I; and 1.3 ± 0.5 μM for rtL180M + rtM204V. Resistance factors (mutant IC50 / wild-type IC50) were 0.7, 2.1, and 0.6, respectively, indicating no significant cross-resistance. [1]
Comparison with AT-61: AT-130 was found to be more potent than its congener AT-61 on a molar basis. AT-61 had IC50 values ranging from 19-27 μM and IC90 values from 63-97 μM against the same HBV variants. [1]
Cytotoxicity Testing: AT-130 was tested for cytotoxicity in HepG2 and Huh-7 human hepatic cell lines using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) cleavage assay after 7 days of continuous exposure. No measurable toxicity was observed at concentrations up to 250 μM, the highest concentration tested. [1]

Cell Culture and Transduction: HepG2 cells were cultured and transduced with recombinant baculoviruses encoding either wild-type HBV or mutant HBV (rtL180M, rtM204I, or rtL180M+rtM204V) at a multiplicity of infection of 50 PFU/cell. Transduction was performed as previously described. [1]
Drug Treatment: Stock solutions of AT-130 were prepared in dimethyl sulfoxide (DMSO) and freshly diluted in culture media. The final DMSO concentration was always <1% (v/v), which has no measurable effect on HBV replication. Beginning immediately after transduction, cells were continuously exposed to five different concentrations of AT-130 for 7 days. Culture media were changed on days 2, 4, and 6 post-transduction. [1]
HBV Replication Analysis: On day 7 post-transduction, cells were harvested. Replicating viral DNA was extracted from cytoplasmic core particles and analyzed by Southern hybridization and autoradiography. Image densities from suitably exposed autoradiographs were measured by computer-assisted densitometry. The amount of viral replication in drug-treated samples was expressed as a percentage of replication in drug-free controls. [1]
Cytotoxicity Assay: HepG2 and Huh-7 cells were continuously exposed to AT-130 for 7 days. Cellular toxicity was assayed colorimetrically by MTT cleavage. [1]
Dose-Response Analysis: Where possible, logistic dose-response curves described by the equation y = a / [1 + (x/b)^(-c)] were fitted to each dataset. Drug concentrations that inhibited replication by 50% (IC50) and 90% (IC90) were estimated from individual dose-response curves. Curve fit parameters and coefficients of determination (r²) were calculated (see Table 1). [1]

[1]. Phenylpropenamide derivatives AT-61 and AT-130 inhibit replication of wild-type and lamivudine-resistant strains of hepatitis B virus in vitro. Antimicrob Agents Chemother. 2002 Sep;46(9):3057-60.

[2]. Phenylpropenamide derivatives as inhibitors of hepatitis B virus replication. Bioorg Med Chem Lett. 2000 Dec 4;10(23):2687-90.

[3]. The phenylpropenamide derivative AT-130 blocks HBV replication at the level of viral RNA packaging. Antiviral Res. 2007 Nov;76(2):168-77.

Background: AT-130 is a phenylpropenamide derivative, a congener of AT-61. Its chemical structure differs from AT-61 in that ring A contains an ortho-methoxy substituent, ring B contains a para-nitroso substituent, and a bromine atom replaces the chlorine present in AT-61 (see Fig. 1 in the original paper). [1]
Mechanism of Action (Proposed): While the exact mechanism is unknown, AT-130 appears to inhibit HBV replication through a mechanism different from nucleoside analogues like lamivudine. Previous studies with AT-61 suggested that phenylpropenamide derivatives may inhibit the packaging of pregenomic viral RNA into cytoplasmic core particles. [1]
Advantage Over Nucleoside Analogues: AT-130 retains activity against HBV strains that are resistant to lamivudine (rtL180M, rtM204I, and rtL180M+rtM204V mutants). This lack of cross-resistance makes it a promising candidate for combination therapy with nucleoside/nucleotide analogs to prevent or overcome drug resistance. [1]
Structural Features: The compound is a low-molecular-weight nonnucleoside inhibitor, which distinguishes it from the nucleoside analog class of HBV drugs. [1]
Clinical Potential: The authors suggest that the favorable activity profile against drug-resistant HBV justifies further development of AT-130 or its derivatives for eventual clinical use, possibly in combination with other anti-HBV drugs. [1]

C22H22BRN3O5

488.3312

487.074

211364-06-6

3002812

White to off-white solid powder

4

1

5

5

31

694

0

COC1=CC=CC=C1/C(=C(/C(=O)N2CCCCC2)\NC(=O)C3=CC=C(C=C3)[N+](=O)[O-])/Br

OQIUTYABZMBBME-FMQUCBEESA-N

InChI=1S/C22H22BrN3O5/c1-31-18-8-4-3-7-17(18)19(23)20(22(28)25-13-5-2-6-14-25)24-21(27)15-9-11-16(12-10-15)26(29)30/h3-4,7-12H,2,5-6,13-14H2,1H3,(H,24,27)/b20-19+

N-[(E)-1-bromo-1-(2-methoxyphenyl)-3-oxo-3-piperidin-1-ylprop-1-en-2-yl]-4-nitrobenzamide

AT-130 AT130 AT 130

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~25 mg/mL (~51.19 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.12 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.

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 (Please use freshly prepared in vivo formulations for optimal results.)