The novel compound Z060228 inhibits assembly of the HBV capsid.
OBJECTIVE
The effective anti-HBV drugs on the market are mainly immunomodulators or nucleoside analogs. The uses of INF-α and lamivudine (3TC) are considerably limited by their low response rate, side effects, drug resistance and HBV recurrence. Thus, new mechanism-based drugs remain in urgent need. This study aimed to investigate the anti-HBV effects of the novel compound Z060228 and to confirm its anti-HBV mechanisms.


METHODS
HepG2.2.15 cells and HBV-transgenic mice were used to evaluate the anti-HBV activity of Z060228. Conformational changes of the capsid structure induced by Z060228 were detected with high-resolution electron microscopy (EM), size-exclusion chromatography (SEC), and atomic force microscopy (AFM).


RESULTS
The HBV DNA replication in the supernatants of the HepG2.2.15 cells was effectively inhibited by Z060228 and Bay41-4109. In the liver of HBV-transgenic mice, the HBcAg content was significantly decreased and HBV DNA replication was also inhibited after high-dose (30 mg/kg) Z060228 treatment. Z060228 and Bay41-4109 exhibited similar effects on the self-assembly of Cp149. SEC data revealed that Z060228 altered the equilibrium (a state of stability) of Cp149 assembly. EM data further demonstrated that Z060228 could prevent Cp149 from self-assembling to the correct core particles. Additionally, AFM results showed that a low concentration of Z060228 caused Cp149 syncretizing, whereas a high concentration caused Cp149 to polymerize.


CONCLUSIONS
Z060228 was demonstrated to be a potential capsid targeting anti-HBV drug candidate. The methods employed here could be used as a general strategy to study mechanisms of self-assembling protein-targeted drugs.
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China; Department of Radiation Toxicology and Oncology, Beijing Key Laboratory for Radiobiology, Beijing Institute of Radiation Medicine, Beijing 100850, People's Republic of China.
Hua
H
Guan
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China.
Guoming
G
Zhao
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China.
Wei
W
Chen
Peking University, Beijing 100871, People's Republic of China.
Guoyi
G
Wu
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China.
Hongying
H
Liu
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China.
Xingkai
X
Jiang
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China. Electronic address: lisong_amms@yahoo.cn.
Song
S
Li
Beijing Institute of Pharmacology and Toxicology, Beijing 100850, People's Republic of China. Electronic address: wangll63@126.com.
Li-li
LL
Wang
D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use*; D002213:Capsid / Q000187:drug effects* / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects; D056945:Hep G2 Cells; D006509:Hepatitis B / Q000188:drug therapy* / Q000235:genetics; D006515:Hepatitis B virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D008099:Liver / Q000821:virology*; D008297:Male; D008822:Mice, Transgenic; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents
Capsid; Cp149; HAPs; HBV; Z060228