Anti-HBV Activity Of Isopropylidene Sedoheptuiosan

PARTⅠ: ANTIVIRAL ACTIVITY OF ISOPROPYLIDENE SEDOHEPTUIOSAN IN HEPG2.2.15 CELL LINEObjectives: To investigate the antiviral activity of isopropylidene sedoheptuiosan in vitro in HepG2.2.15 cell line.Methods: HBV genomic DNA transfected HepG2.2.15 cell lines were treated with isopropylidene sedoheptuiosan of concentration 10μg/ml, 40μg/ml, 100μg/ml for 72h, entecavir (10μg/ml) as positive control group and no-drug-containing culture media as negative control group. Culture supernatants at 24h,48h and 72h were harvested to measure the levels of HBsAg and HBeAg using micro-article enzyme immune assay(MEIA). HBV-DNA levels both in those supernatants and in the cells were determined by fluorescence quantative polymerase chain reaction (PCR).Methyl Thiazolyl Tetrazolium assay was used to evaluate the cytotoxicity of isopropylidene sedoheptuiosan to the experimental cells.Results: The survival rates of HepG2.2.15 cell lines in 10μg/ml, 40μg/ml, 100μg/ml group were 93.09%, 91.12% and 87.03%(P>0.05), respectively;The levels of HBsAg, HBeAg and HBV-DNA in culture supernatant at 24h, 48h, 72h and levels of HBV-DNA in the cells were lower in treatment groups than those in negative control group. But no significant differences were observed among those groups. The suppressive rates of drug on HBsAg, HBeAg and HBV-DNA increased with the enlengthing duration of treatment, and the suppressive rates in high-dose group were higher than those in low-dose ones, but no significant differences were observed among them. However, enticavir can obviously down regulate HBV load both in the supernatant and in the cells.Conclusion: isopropylidene sedoheptuiosan with the concentration of 10μg/ml, 40μg/ml and 100μg/ml had no cytotoxicity in HepG2.2.15 cell line, and also no significant antiviral efficacy on HBV was observed.PARTⅡTHE SAFETY AND EFFICACY OF THE TREATMENT WITH ISOPROPYLIDENE SEDOHEPTUIOSAN IN HBEAG POSITIVE CHRONIC HEPATITIS B(CHB) PATIENTS Objectives: To investigate the safety and efficacy of isopropylidene sedoheptuiosan in treating HBeAg positive chronic hepatitis B (CHB) patients, and to explore the propable antiviral mechanism.Patients and Methods: HBeAg positive CHB patients with elevated alanine aminotransferase(ALT) and HBV-DNA level﹥1.0×105copies/ml participated in the clinical study. Those patients were randomly distributed to the treatment groups (800mg, 400mg, and 200mg) and the control group with the ratio of 1:1:1:1. Patients in treatment groups received isopropylidene sedoheptuiosan of various dosage and those in control group received mannitol.The duration was 12 weeks. Virological, biochemical and serological responses, adverse effects and levels of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α) and interleukine-2 in serum were detected before and after treatment.Result: The virologic response rates in 800mg, 400mg and 200mg groups were 18.75%, 12.5% and 12.5%, respectively. The differences were no statistical significance.There were no signigicant differences in the median decreased HBV-DNA level after treatment among treatment groups and control group. Significant differences of log (HBV-DNA) before and after 12 weeks treatment were observed in 800mg group. No significant differences were observed both in HBeAg seroconversion rate and in ALT normalization rate among those groups. No adverse effects were observed in 800mg group, and only one severe adverse event occurred in control group.There were no differences in serum IFN-γlevels before and after treatment in every group. Obvious increases in the levels of TNF-αand IL-2 at 12 week after treatment were detected in 800mg group(P=0.015,0.023), while no obvious increases were observed among other groups.We analyzed the differences between the levels of IFN-γ,TNF-αand IL-2 before and after 12 weeks treatment among treatment groups and control group.Results showed that there were significant differences between 800mg group and control group (P=0.031, 0.005, and 0.041).Conclusion: Isopropylidene sedoheptuiosan of 800mg could significantly decreased the serum HBV-DNA load probably through activating host immune response. It showed some antiviral efficiacy and good security in HBeAg positive CHB patients. However, this conclusion needs to be checked up with large scale clinical trials.