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689
S-ANTIGEN TRAFFIC-INHIBITING OLIGONUCLEOTIDE
POLYMERS (STOPS) CAN EFFECTIVELY INHIBIT HEPATITIS B
SURFACE ANTIGEN (HBsAg) SECRETION FROM HEPATITIS B
VIRUS (HBV) CELL LINES
Jin Hong, Rajendra Pandey, Vivek K. Rajwanshi, Hua
Tan, Yuchun Nie, Cheng Kao, Suping Ren, John Cortez,
Dinah Misner, Sushmita Chanda, David B. Smith, Julian
Symons, Lawrence M Blatt and Leonid N. Beigelman, Aligos
Therapeutics
Background: Chronic Hepatitis B (CHB) is a global public
health problem, affecting 300 million people Current standard
of care can effectively inhibit viral DNA replication but fails to
reduce HBsAg that suppresses the human immune system
and prevents the attainment of “functional cure” Nucleic
acid polymers (NAPs) have been reported to significantly
reduce circulating HBsAg in CHB patients when given as
monotherapy1 and in combination therapy2. We have studied
oligonucleotides that can inhibit HBsAg secretion and have
identified STOPs that share structural similarity with NAPs but
contain several novel chemical features, providing enhanced
potency in several HBV cell lines. Methods: STOPs were
synthesized on an ABI 394 and Expedite 8909 synthesizers
using standard phosphoramidite chemistry Compounds
were profiled in the HepG2.2.15 cell line and HepG2-NTCP
live HBV infection system. In HepG2.2.15, compounds were
administered by transfection using RNAiMAX and released
HBsAg was measured by ELISA 6 days post transfection
HepG2-NTCP cells were infected with live HBV at 200 moi (ge)
and 5 days later, STOPs were added and HBsAg measured as
for HepG2 2 15 cells Results: In vitro profiling identified ALG-
10093, as a potent STOP with EC50 values of 2 5 nM and 2 1
nM in HBsAg release assays from HepG2 2 15 and HepG2-
NTCP/live HBV systems respectively. This was approximately
>100-fold more potent than the lead NAP currently in clinical
trials In probing the mechanism of action of STOPs, we
found reduced intracellular HBsAg levels similar to secreted
HBsAg, suggesting that HBsAg was degraded intracellularly
There was no effect on intracellular HBV RNA, excluding the
RNaseH pathway as an HBV RNA degradation mechanism
for STOPs HBeAg secretion was inhibited concomitantly
with HBsAg, however, extracellular HBV DNA and RNA were
not inhibited, indicating that STOPs do not affect capsid
assembly. Biochemical studies failed to find binding of STOPs
to HBV antigens and demonstrated interactions with multiple
host proteins Conclusion: STOPs such as ALG-10093
are a class of very potent oligonucleotides that can reduce
HBsAg secretion by potentially affecting protein trafficking
from the infected cell STOPs have the potential to be used
therapeutically to reduce HBsAg in CHB patients to achieve a
functional cure Further investigation is therefore warranted. |
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发表于 2019-10-30 17:32
