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406 Interferon therapy improves prognosis in patients with HBeAg positive chronic hepatitis B
Category 5a." Viral Hepatitis." Hepatitis B Basic may reflect differences both in the genetic make-up of the virus and the molecular mechanism of HBV morphogenesis in the natural history of HBV infection.
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COMPARATIVE DISPOSITION AND METABOLIC PROFILES OF [14C]REMOFOVIR AND [14ClADEFOVIR DIPIVOXIL IN RAT LIVER AND KIDNEY
C-C. Lin, N. Zhu, D. Lourenco, L-T. Yeh. Drug Development, Valeant Pharmaceuticals International, Costa Mesa, CA, USA Background: Adefovir dipivoxil, an estevase-activated prodrug of PMEA has a recommended dose of 10rag QD, which is sub-optimal due to dose-limiting nephrotoxicity. Remofovir is a CYP3A4-activated prodrug of PMEA with excellent liver-target properties in vats and improved safety compared to adefovir dipivoxil in a one-month monkey toxicology study. Th~ls treatment with remofovir may result in improved efficacy and lower potential for nephrotoxicity than adefovir dipivoxil. Aim: To evaluate disposition and metabolic profile of remofovir and adefovir dipivoxil in rat liver and kidney following oral administration. Methods: Following an overnight fast, vats received ovally 30mg/kg of [14C]remofovir or [14C]adefovir dipivoxil. At 3 hours, liver and kidney were collected, sohibilized with KOH, decolorized with H202 and neutralized, followed by radioactivity determination. Liver and kidney samples were also extracted with perchloric acid, homogenized and centrifuged, followed by" metabolic profiling using a HPLC procedure with DEAE column. Results: Oral dosing of adefovir dipivoxil resulted in a radioactivity level of 78.3 ~tM and 12.9~tM in !ddney and liver, respectivelg In contrast, oral dosing of remofovir resulted in a radioactivity level of 13.6 ~tM and 41.3 tIM in kidney and liver, respectively. Following single oral dosing of renmfovir, PMEA, PMEAp and PMEApp accounted for 36.2, 34.0 and 29.8% of total radioactivity in the liver, respectively, and accounted for 35.3, 49.4 and 7.4% of total radioactivity in the kidney, respectivel3~ In addition, renmfovir was also detected in kidney and accounted for 7.8% of kithley radioacti,Aty. Following single oral dosing of adefovir dipivoxil, PMEA, PMEAp and PMEApp accounted for 43.4, 31.3 and 25.3% of total radioactivity in the liver, respectively, and accounted for 29.9, 33.8 and 36.4% of total radioactivity in the kidney, respectively. Conclusions: Administration of remofovir resulted in a 3-fold higher radioactivity level in the liver but only 1/6 in the kidneys, compared to the levels afier dosing with adefovir dipivoxil, tn vat kidney and liver, PMEA, PMEAp and PMEApp were the major radioactive peaks, except that unchanged remofovir was detected in rat kidney following dosing of [14C]remofovir.
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INTERFERON T H E R A P Y IMPROVES P R O G N O S I S IN PATIENTS WITH HBeAg POSITIVE CHRONIC HEPATITIS B
S.M. Lin 1, R.N. Chien2, I.S. Sheen 1, C.M. Lee 1, M i . Yu 1, C.M. Chu 1, Y.E Liaw I . 1Liver Research Unit, Chang Gung Memorial Hospital,
Taipei, Taiwan; 2Division of Hepatology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan Background: The effectiveness of interferon (IFN) on the prognosis in patients with chronic hepatitis B is controversial. Methods: A total of 466 patients (233 in IFN group, 233 in well-matched control group) were enrolled. Results: Seventy one (30°/5) patients in IFN group while 32 (14%) controls (P <0.001) were responders. One hundred and fifteen (49.4%) patients in IFN group and 86 (36.9%) controls (P<0.01) were seroconverters (sustained responders). The cumulative incidence of cirrhosis at 11 years were 33.7% in control group and 17.8% in IFN group (P=0.041); 45.3% in control-nonseroconverter subgroup and higher than 13.5% in controlseroconverter subgroup (P = 0.031), than 21.6% in IFN-nonseroconverter
149
subgroup (P=0.065), and than 10.4% in IFN-seroconverter subgroup (P = 0.023). The cumulative incidence of hepatocellular carcinoma (HCC) at 11 years was 12.5% in control group and 2.7% in IFN group (P = 0.011); 17.9% in control-nonseroconverter subgroup and higher than 3.5% in control-seroconverter subgroup (P=0.009), than 0% in IFNseroconverter subgroup (P=0.003), than 5.2% in IFN-nonseroconverter subgroup (P=0.014); the rate also higher in IFN-nonseroconverter subgroup than IFN-seroconverter subgroup (P= 0.019). The cumulative rate of survival at 11 years were 52.7% in control group and 97.5% in IFN group(P=0.003); 33.7% in control-nonseroconverter subgroup and higher than 96.3% in control-seroconverter subgroup (P=0.034), than 100% in IFN-seroconverter subgroup (P =0.001), and than 93.8% in IFNnonseroconverter subgroup (P = 0.026). Conclusion: IFN therapy improves prog-nosis in terms of seroconversion, redudng cirrhosis and HCC development, and prolonging survival in patients with chronic hepatitis B, particularly in seroconverters. OF ISOLATED ANTIBODIES TO 41407W•PERSISTENCE O O D C H U C K HEPATITIS VIRUS CORE ANTIGEN
IS
INDICATIVE OF OCCULT INFECTION
C.S. Coffin, T.N.Q. Pham, RM. Mulrooney, N.D. Churchill, T.I. Michalak.
Molecular Virology and Hepatology Research, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, Canada Antibodies against HBV nucleocapsid (anti-HBc) may occur in the absence of symptoms and other serological indicators of the infection. This situation can be encountered following a clinically and serologically unapparent exposure to HBV or after recovery from hepatitis B. In this study, woodchucks inoculated with woodehnck hepatitis virus (WHV) were investigated to determine a relationship between detection of isolated anticore (also termed anti-core alone) and the molecular status of virus replication in a primary WHV snrface antigen (WHsAg)-negative infection or long after resolution of WHV hepatitis. Parallel samples of sera, peripheral blood mononuclear cells (PBMC) and liver tissue, serially collected for more than 5 years after inoculation with virus, were examined for WHV DNA by PCWnncleic acid hybridization (PCPdNAH) assays. Sera were also tested for WHV DNA after DNase treatment and for WHV DNA and WHsAg after concentration in sucrose. Liver biopsies and PBMC were examined for WHV cccDNA and viral RNA transcripts by PCR-based techniques to assess virus replication status. The study shows that anti-core antibodies existing in the absence of other serological markers are a reliable indicator of occult WHV infection. This state can be accompanied by traces of circulating particles behaving as intact virions and by intermittent minimal to mild liver iTtflammation. The findings suggest that the long-term presence of anti-core antibodies alone is a consequence of sustained restimulation of the immune system by hepadnavirus nucleocapsid produced during low-level continued viral assembly. A high degree o f virological and pathogenic compatibility between HBV and WHV suggests that the occurrence of isolated anti-HBc could be important in identifying occult HBV persistence in humans.
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SINGLE-CHAIN Fv (scFc) ANTIBODIES FROM THE CDR3 VH MUTANT LIBRARY INHIBITS THE BINDING OF PRES1 OF HEPATITIS B VIRUS TO THE LIVER ORIGIN CELL LINES
N.J. Park 1, Y.J. Jeong 2, S.R. Jee I , E.T. Park t , Y.J. Lee t, S.H. Lee 1, S.Y. Seol 1, Y.Y. Lee2, S.G. Park 2, I.H. Choi 2, J.M. Chung t . 1Department
of Internal Medicine, Collage of Medicine, Inje University. Busan, South Korea," 2Department of Microbiology and Immunology, Collage of Medicine, Inje University, Busan, South Korea The complementarity determining regions (CDRs) within the Fv are the regions both mediating a potential antigen binding and being mutated at a
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COMPARATIVE DISPOSITION AND METABOLIC PROFILES OF [14C]REMOFOVIR AND [14ClADEFOVIR DIPIVOXIL IN RAT LIVER AND KIDNEY
C-C. Lin, N. Zhu, D. Lourenco, L-T. Yeh. Drug Development, Valeant Pharmaceuticals International, Costa Mesa, CA, USA Background: Adefovir dipivoxil, an estevase-activated prodrug of PMEA has a recommended dose of 10rag QD, which is sub-optimal due to dose-limiting nephrotoxicity. Remofovir is a CYP3A4-activated prodrug of PMEA with excellent liver-target properties in vats and improved safety compared to adefovir dipivoxil in a one-month monkey toxicology study. Th~ls treatment with remofovir may result in improved efficacy and lower potential for nephrotoxicity than adefovir dipivoxil. Aim: To evaluate disposition and metabolic profile of remofovir and adefovir dipivoxil in rat liver and kidney following oral administration. Methods: Following an overnight fast, vats received ovally 30mg/kg of [14C]remofovir or [14C]adefovir dipivoxil. At 3 hours, liver and kidney were collected, sohibilized with KOH, decolorized with H202 and neutralized, followed by radioactivity determination. Liver and kidney samples were also extracted with perchloric acid, homogenized and centrifuged, followed by" metabolic profiling using a HPLC procedure with DEAE column. Results: Oral dosing of adefovir dipivoxil resulted in a radioactivity level of 78.3 ~tM and 12.9~tM in !ddney and liver, respectivelg In contrast, oral dosing of remofovir resulted in a radioactivity level of 13.6 ~tM and 41.3 tIM in kidney and liver, respectively. Following single oral dosing of renmfovir, PMEA, PMEAp and PMEApp accounted for 36.2, 34.0 and 29.8% of total radioactivity in the liver, respectively, and accounted for 35.3, 49.4 and 7.4% of total radioactivity in the kidney, respectivel3~ In addition, renmfovir was also detected in kidney and accounted for 7.8% of kithley radioacti,Aty. Following single oral dosing of adefovir dipivoxil, PMEA, PMEAp and PMEApp accounted for 43.4, 31.3 and 25.3% of total radioactivity in the liver, respectively, and accounted for 29.9, 33.8 and 36.4% of total radioactivity in the kidney, respectively. Conclusions: Administration of remofovir resulted in a 3-fold higher radioactivity level in the liver but only 1/6 in the kidneys, compared to the levels afier dosing with adefovir dipivoxil, tn vat kidney and liver, PMEA, PMEAp and PMEApp were the major radioactive peaks, except that unchanged remofovir was detected in rat kidney following dosing of [14C]remofovir.
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INTERFERON T H E R A P Y IMPROVES P R O G N O S I S IN PATIENTS WITH HBeAg POSITIVE CHRONIC HEPATITIS B
S.M. Lin 1, R.N. Chien2, I.S. Sheen 1, C.M. Lee 1, M i . Yu 1, C.M. Chu 1, Y.E Liaw I . 1Liver Research Unit, Chang Gung Memorial Hospital,
Taipei, Taiwan; 2Division of Hepatology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan Background: The effectiveness of interferon (IFN) on the prognosis in patients with chronic hepatitis B is controversial. Methods: A total of 466 patients (233 in IFN group, 233 in well-matched control group) were enrolled. Results: Seventy one (30°/5) patients in IFN group while 32 (14%) controls (P <0.001) were responders. One hundred and fifteen (49.4%) patients in IFN group and 86 (36.9%) controls (P<0.01) were seroconverters (sustained responders). The cumulative incidence of cirrhosis at 11 years were 33.7% in control group and 17.8% in IFN group (P=0.041); 45.3% in control-nonseroconverter subgroup and higher than 13.5% in controlseroconverter subgroup (P = 0.031), than 21.6% in IFN-nonseroconverter
149
subgroup (P=0.065), and than 10.4% in IFN-seroconverter subgroup (P = 0.023). The cumulative incidence of hepatocellular carcinoma (HCC) at 11 years was 12.5% in control group and 2.7% in IFN group (P = 0.011); 17.9% in control-nonseroconverter subgroup and higher than 3.5% in control-seroconverter subgroup (P=0.009), than 0% in IFNseroconverter subgroup (P=0.003), than 5.2% in IFN-nonseroconverter subgroup (P=0.014); the rate also higher in IFN-nonseroconverter subgroup than IFN-seroconverter subgroup (P= 0.019). The cumulative rate of survival at 11 years were 52.7% in control group and 97.5% in IFN group(P=0.003); 33.7% in control-nonseroconverter subgroup and higher than 96.3% in control-seroconverter subgroup (P=0.034), than 100% in IFN-seroconverter subgroup (P =0.001), and than 93.8% in IFNnonseroconverter subgroup (P = 0.026). Conclusion: IFN therapy improves prog-nosis in terms of seroconversion, redudng cirrhosis and HCC development, and prolonging survival in patients with chronic hepatitis B, particularly in seroconverters. OF ISOLATED ANTIBODIES TO 41407W•PERSISTENCE O O D C H U C K HEPATITIS VIRUS CORE ANTIGEN
IS
INDICATIVE OF OCCULT INFECTION
C.S. Coffin, T.N.Q. Pham, RM. Mulrooney, N.D. Churchill, T.I. Michalak.
Molecular Virology and Hepatology Research, Faculty of Medicine, Memorial University of Newfoundland, St. John's, NL, Canada Antibodies against HBV nucleocapsid (anti-HBc) may occur in the absence of symptoms and other serological indicators of the infection. This situation can be encountered following a clinically and serologically unapparent exposure to HBV or after recovery from hepatitis B. In this study, woodchucks inoculated with woodehnck hepatitis virus (WHV) were investigated to determine a relationship between detection of isolated anticore (also termed anti-core alone) and the molecular status of virus replication in a primary WHV snrface antigen (WHsAg)-negative infection or long after resolution of WHV hepatitis. Parallel samples of sera, peripheral blood mononuclear cells (PBMC) and liver tissue, serially collected for more than 5 years after inoculation with virus, were examined for WHV DNA by PCWnncleic acid hybridization (PCPdNAH) assays. Sera were also tested for WHV DNA after DNase treatment and for WHV DNA and WHsAg after concentration in sucrose. Liver biopsies and PBMC were examined for WHV cccDNA and viral RNA transcripts by PCR-based techniques to assess virus replication status. The study shows that anti-core antibodies existing in the absence of other serological markers are a reliable indicator of occult WHV infection. This state can be accompanied by traces of circulating particles behaving as intact virions and by intermittent minimal to mild liver iTtflammation. The findings suggest that the long-term presence of anti-core antibodies alone is a consequence of sustained restimulation of the immune system by hepadnavirus nucleocapsid produced during low-level continued viral assembly. A high degree o f virological and pathogenic compatibility between HBV and WHV suggests that the occurrence of isolated anti-HBc could be important in identifying occult HBV persistence in humans.
[
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SINGLE-CHAIN Fv (scFc) ANTIBODIES FROM THE CDR3 VH MUTANT LIBRARY INHIBITS THE BINDING OF PRES1 OF HEPATITIS B VIRUS TO THE LIVER ORIGIN CELL LINES
N.J. Park 1, Y.J. Jeong 2, S.R. Jee I , E.T. Park t , Y.J. Lee t, S.H. Lee 1, S.Y. Seol 1, Y.Y. Lee2, S.G. Park 2, I.H. Choi 2, J.M. Chung t . 1Department
of Internal Medicine, Collage of Medicine, Inje University. Busan, South Korea," 2Department of Microbiology and Immunology, Collage of Medicine, Inje University, Busan, South Korea The complementarity determining regions (CDRs) within the Fv are the regions both mediating a potential antigen binding and being mutated at a