BARACLUDE
®
(entecavir) BARACLUDE
®
(entecavir)
entecavir was not antagonistic to the cell culture anti-HIV activity of these six NRTIs or
emtricitabine at concentrations greater than 100 times the C
max
of entecavir using the
1 mg dose.
Antiviral Activity Against HIV
A comprehensive analysis of the inhibitory activity of entecavir against a panel of laboratory
and clinical HIV type 1 (HIV-1) isolates using a variety of cells and assay conditions yielded
EC
50
values ranging from 0.026 to >10 µM; the lower EC
50
values were observed when
decreased levels of virus were used in the assay. In cell culture, entecavir selected for an
M184I substitution in HIV reverse transcriptase at micromolar concentrations, confirming
inhibitory pressure at high entecavir concentrations. HIV variants containing the M184V
substitution showed loss of susceptibility to entecavir.
Resistance
In Cell Culture
In cell-based assays, 8- to 30-fold reductions in entecavir phenotypic susceptibility were
observed for lamivudine-resistant strains. Further reductions (>70-fold) in entecavir
phenotypic susceptibility required the presence of amino acid substitutions rtM204I/V
with or without rtL180M along with additional substitutions at residues rtT184, rtS202,
or rtM250, or a combination of these substitutions with or without an rtI169 substitution
in the HBV reverse transcriptase. Lamivudine-resistant strains harboring rtL180M
plus rtM204V in combination with the amino acid substitution rtA181C conferred
16- to 122-fold reductions in entecavir phenotypic susceptibility.
Clinical Studies
Nucleoside-inhibitor-naïve subjects: Genotypic evaluations were performed on evaluable
samples (>300 copies/mL serum HBV DNA) from 562 subjects who were treated
with BARACLUDE for up to 96 weeks in nucleoside-inhibitor-naïve studies (AI463022,
AI463027, and rollover study AI463901). By Week 96, evidence of emerging amino acid
substitution rtS202G with rtL180M and rtM204V substitutions was detected in the HBV of
2 subjects (2/562=<1%), and 1 of them experienced virologic rebound (≥1 log
10
increase
above nadir). In addition, emerging amino acid substitutions at rtM204I/V and rtL80I,
rtV173L, or rtL180M, which conferred decreased phenotypic susceptibility to entecavir in
the absence of rtT184, rtS202, or rtM250 changes, were detected in the HBV of 3 subjects
(3/562=<1%) who experienced virologic rebound. For subjects who continued treatment
beyond 48 weeks, 75% (202/269) had HBV DNA <300 copies/mL at end of dosing
(up to 96 weeks).
HBeAg-positive (n=243) and -negative (n=39) treatment-naïve subjects who failed to
achieve the study-defined complete response by 96 weeks were offered continued
entecavir treatment in a rollover study. Complete response for HBeAg-positive was
<0.7 MEq/mL (approximately 7 × 10
5
copies/mL) serum HBV DNA and HBeAg loss
and, for HBeAg-negative was <0.7 MEq/mL HBV DNA and ALT normalization. Subjects
received 1 mg entecavir once daily for up to an additional 144 weeks. Of these
282 subjects, 141 HBeAg-positive and 8 HBeAg-negative subjects entered the long-term
follow-up rollover study and were evaluated for entecavir resistance. Of the 149 subjects
entering the rollover study, 88% (131/149), 92% (137/149), and 92% (137/149) attained
serum HBV DNA <300 copies/mL by Weeks 144, 192, and 240 (including end of dosing),
respectively. No novel entecavir resistance-associated substitutions were identified in a
comparison of the genotypes of evaluable isolates with their respective baseline isolates.
The cumulative probability of developing rtT184, rtS202, or rtM250 entecavir resistance-
associated substitutions (in the presence of rtL180M and rtM204V substitutions) at
Weeks 48, 96, 144, 192, and 240 was 0.2%, 0.5%, 1.2%, 1.2%, and 1.2%, respectively.
Lamivudine-refractory subjects: Genotypic evaluations were performed on evaluable
samples from 190 subjects treated with BARACLUDE for up to 96 weeks in studies of
lamivudine-refractory HBV (AI463026, AI463014, AI463015, and rollover study AI463901).
By Week 96, resistance-associated amino acid substitutions at rtT184, rtS202, or
rtM250, with or without rtI169 changes, in the presence of amino acid substitutions
rtM204I/V with or without rtL80V, rtV173L/M, or rtL180M emerged in the HBV from
22 subjects (22/190=12%), 16 of whom experienced virologic rebound (≥1 log
10
increase
above nadir) and 4 of whom were never suppressed <300 copies/mL. The HBV from 4
of these subjects had entecavir resistance substitutions at baseline and acquired further
changes on entecavir treatment. In addition to the 22 subjects, 3 subjects experienced
virologic rebound with the emergence of rtM204I/V and rtL80V, rtV173L/M, or rtL180M.
For isolates from subjects who experienced virologic rebound with the emergence
of resistance-associated substitutions (n=19), the median fold-change in entecavir
EC
50
values from reference was 19-fold at baseline and 106-fold at the time of virologic
rebound. For subjects who continued treatment beyond 48 weeks, 40% (31/77) had
HBV DNA <300 copies/mL at end of dosing (up to 96 weeks).
Lamivudine-refractory subjects (n=157) who failed to achieve the study-defined complete
response by Week 96 were offered continued entecavir treatment. Subjects received 1 mg
entecavir once daily for up to an additional 144 weeks. Of these subjects, 80 subjects
entered the long-term follow-up study and were evaluated for entecavir resistance.
By Weeks 144, 192, and 240 (including end of dosing), 34% (27/80), 35% (28/80), and
36% (29/80), respectively, attained HBV DNA <300 copies/mL. The cumulative probability
of developing rtT184, rtS202, or rtM250 entecavir resistance-associated substitutions (in
the presence of rtM204I/V with or without rtL180M substitutions) at Weeks 48, 96, 144,
192, and 240 was 6.2%, 15%, 36.3%, 46.6%, and 51.5%, respectively. The HBV of 6
subjects developed rtA181C/G/S/T amino acid substitutions while receiving entecavir, and
of these, 4 developed entecavir resistance-associated substitutions at rtT184, rtS202, or
rtM250 and 1 had an rtT184S substitution at baseline. Of 7 subjects whose HBV had an
rtA181 substitution at baseline, 2 also had substitutions at rtT184, rtS202, or rtM250 at
baseline and another 2 developed them while on treatment with entecavir.
In a post-approval integrated analysis of entecavir resistance data from 17 Phase 2
and 3 clinical trials, an emergent entecavir resistance-associated substitution rtA181C
was detected in 5 out of 1461 (0.3%) subjects during treatment with entecavir. This
substitution was detected only in the presence of lamivudine resistance-associated
substitutions rtL180M plus rtM204V.
Cross-resistance
Cross-resistance has been observed among HBV nucleoside analogue inhibitors.
In cell-based assays, entecavir had 8- to 30-fold less inhibition of HBV DNA synthesis
for HBV containing lamivudine and telbivudine resistance-associated substitutions
rtM204I/V with or without rtL180M than for wild-type HBV. Substitutions rtM204I/V with
or without rtL80I/V, rtV173L, or rtL180M, which are associated with lamivudine and
telbivudine resistance, also confer decreased phenotypic susceptibility to entecavir. The
efficacy of entecavir against HBV harboring adefovir resistance-associated substitutions
has not been established in clinical trials. HBV isolates from lamivudine-refractory subjects
failing entecavir therapy were susceptible in cell culture to adefovir but remained resistant
to lamivudine. Recombinant HBV genomes encoding adefovir resistance-associated
substitutions at either rtA181V or rtN236T had 1.1- or 0.3-fold shifts in susceptibility to
entecavir in cell culture, respectively.
13 NONCLINICAL TOXICOLOGY
13.1 Carcinogenesis, Mutagenesis, Impairment of Fertility
Carcinogenesis
Long-term oral carcinogenicity studies of entecavir in mice and rats were carried out
at exposures up to approximately 42 times (mice) and 35 times (rats) those observed
in humans at the highest recommended dose of 1 mg/day. In mouse and rat studies,
entecavir was positive for carcinogenic findings. It is not known how predictive the results
of rodent carcinogenicity studies may be for humans [see Adverse Reactions (6.2)].
In mice, lung adenomas were increased in males and females at exposures 3 and
40 times those in humans. Lung carcinomas in both male and female mice were
increased at exposures 40 times those in humans. Combined lung adenomas and
carcinomas were increased in male mice at exposures 3 times and in female mice at
exposures 40 times those in humans. Tumor development was preceded by pneumocyte
proliferation in the lung, which was not observed in rats, dogs, or monkeys administered
entecavir, supporting the conclusion that lung tumors in mice may be a species-specific
event. Hepatocellular carcinomas were increased in males and combined liver adenomas
and carcinomas were also increased at exposures 42 times those in humans. Vascular
tumors in female mice (hemangiomas of ovaries and uterus and hemangiosarcomas of
spleen) were increased at exposures 40 times those in humans. In rats, hepatocellular
adenomas were increased in females at exposures 24 times those in humans; combined
adenomas and carcinomas were also increased in females at exposures 24 times those
in humans. Brain gliomas were induced in both males and females at exposures 35 and
24 times those in humans. Skin fibromas were induced in females at exposures 4 times
those in humans.
Mutagenesis
Entecavir was clastogenic to human lymphocyte cultures. Entecavir was not mutagenic in
the Ames bacterial reverse mutation assay using S. typhimurium and E. coli strains in the
presence or absence of metabolic activation, a mammalian-cell gene mutation assay, and
a transformation assay with Syrian hamster embryo cells. Entecavir was also negative in
an oral micronucleus study and an oral DNA repair study in rats.
Impairment of Fertility
In reproductive toxicology studies, in which animals were administered entecavir at up
to 30 mg/kg for up to 4 weeks, no evidence of impaired fertility was seen in male or
female rats at systemic exposures greater than 90 times those achieved in humans
at the highest recommended dose of 1 mg/day. In rodent and dog toxicology studies,
seminiferous tubular degeneration was observed at exposures 35 times or greater than
those achieved in humans. No testicular changes were evident in monkeys.
14 CLINICAL STUDIES
14.1 Outcomes in Adults
At 48 Weeks
The safety and efficacy of BARACLUDE in adults were evaluated in three Phase 3
active-controlled trials. These studies included 1633 subjects 16 years of age or older
with chronic hepatitis B virus infection (serum HBsAg-positive for at least 6 months)
accompanied by evidence of viral replication (detectable serum HBV DNA, as measured
by the bDNA hybridization or PCR assay). Subjects had persistently elevated ALT levels at
least 1.3 times ULN and chronic inflammation on liver biopsy compatible with a diagnosis
of chronic viral hepatitis. The safety and efficacy of BARACLUDE were also evaluated in a
study of 191 HBV-infected subjects with decompensated liver disease and in a study of
68 subjects co-infected with HBV and HIV.
Nucleoside-inhibitor-naïve Subjects with Compensated Liver Disease
HBeAg-positive: Study AI463022 was a multinational, randomized, double-blind study of
BARACLUDE 0.5 mg once daily versus lamivudine 100 mg once daily for a minimum of
52 weeks in 709 (of 715 randomized) nucleoside-inhibitor-naïve subjects with chronic
hepatitis B virus infection, compensated liver disease, and detectable HBeAg. The mean
age of subjects was 35 years, 75% were male, 57% were Asian, 40% were Caucasian,
and 13% had previously received interferon-α. At baseline, subjects had a mean