Hepatitis: Zhou Y

Li M, Zhou Y, Feng G, Su SB. · Institute of Inflammation and Immune Diseases, Shantou University Medical College, Shantou 515041, China. · Curr Mol Med. · Pubmed #19355917 No free full text.

Abstract: Toll-like receptors (TLRs) form a large family of pattern recognition receptors with at least 11 members in human and 13 in mouse. TLRs recognize a wide variety of microbial components and potential host-derived agonists that have emerged as key mediators of innate immunity. TLR signaling also plays an important role in the activation of the adaptive immune system by inducing proinflammatory cytokines and upregulating costimulatory molecules of antigen presenting cells. The dysregulation of TLR signaling may cause autoimmunity. This review discusses the contribution of TLR signaling to the initiation and progression of autoimmune diseases, such as rheumatoid arthritis, experimental autoimmune encephalitis, myocarditis, hepatitis, kidney disease, systemic lupus erythematosus, diabetes, obesity, and experimental autoimmune uveitis as well as aging. The involvement of TLR signaling in the pathogenesis of autoimmune diseases may provide novel targets for the development of therapeutics.

Zhou Y, Bartels DJ, Hanzelka BL, Müh U, Wei Y, Chu HM, Tigges AM, Brennan DL, Rao BG, Swenson L, Kwong AD, Lin C. · Vertex Pharmaceuticals Incorporated, 130 Waverly Street, Cambridge, MA 02139, USA. · Antimicrob Agents Chemother. · Pubmed #17938182 links to  free full text

Abstract: In patients chronically infected with hepatitis C virus (HCV) strains of genotype 1, rapid and dramatic antiviral activity has been observed with telaprevir (VX-950), a highly selective and potent inhibitor of the HCV NS3-4A serine protease. HCV variants with substitutions in the NS3 protease domain were observed in some patients during telaprevir dosing. In this study, purified protease domain proteins and reconstituted HCV subgenomic replicons were used for phenotypic characterization of many of these substitutions. V36A/M or T54A substitutions conferred less than eightfold resistance to telaprevir. Variants with double substitutions at Val36 plus Thr54 had approximately 20-fold resistance to telaprevir, and variants with double substitutions at Val36 plus Arg155 or Ala156 had >40-fold resistance to telaprevir. An X-ray structure of the HCV strain H protease domain containing the V36M substitution in a cocomplex with an NS4A cofactor peptide was solved at a 2.4-A resolution. Except for the side chain of Met36, the V36M variant structure is identical to that of the wild-type apoenzyme. The in vitro replication capacity of most variants was significantly lower than that of the wild-type replicon in cells, which is consistent with the impaired in vivo fitness estimated from telaprevir-dosed patients. Finally, the sensitivity of these replicon variants to alpha interferon or ribavirin remained unchanged compared to that of the wild-type.

Sun Z, Wang Y, Fu Q, Zhou Y, Jia S, Du J, Peng J, Wang Y, Yang S, Zhan L. · Beijing Institute of Transfusion Medicine, Beijing, China. · Antivir Ther. · Pubmed #19474473 No free full text.

Abstract: BACKGROUND: The lack of a robust small animal model for hepatitis C virus (HCV) has hindered the development of novel drugs, including internal ribosome entry site (IRES) inhibitors. Phage phiC31 integrase has emerged as a potent tool for achieving long-term gene expression in vivo. This study utilized phiC31 integrase to develop a stable, reproducible and easily accessible HCV IRES mouse model. METHODS: phiC31 integrase plasmid and the reporter vector, HCV-IRES-luciferase expression cassette (containing an attB site), was codelivered to murine livers using high pressure tail vein injection. HCV IRES-dependent translation reflected by luciferase expression was accurately monitored in vivo by bioluminescence imaging. Genomic integration of the transgene was confirmed by partial hepatectomy and nested PCR. An HCV IRES-targeted short hairpin RNA (shRNA) expression plasmid, sh184, was hydrodynamically transfected into mouse liver to study its inhibition efficacy in vivo. RESULTS: phiC31 integrase mediated intramolecular recombination between wild-type attB and attP sites in mice. The expression of luciferase was stable after 30 days post-transfection and remained so for 300 days only in the livers of mice that were coinjected with the integrase-encoding plasmid. Luciferase levels reduced dramatically after hydrodynamic transfection of sh184. CONCLUSIONS: These results indicate that this mouse model provides a powerful tool for accurate and long-term evaluation of potential anti-IRES compounds in vivo.

Wang X, Zhou Y, Sun L, Chen W, Li X, Wang Q, Lin H. · Laboratory of Molecular Pathology, Shaanxi University of Chinese Medicine, Xianyang, Shaanxi 712046, People's Republic of China. · J Proteome Res. · Pubmed #19367718 No free full text.

Abstract: Hepatitis B virus (HBV) infection is a major factor contributing to the development of hepatocellular carcinoma (HCC) in the world. Hepatitis B virus X protein (HBx) has been verified to play an important role in hepatocarcinogenesis. Heat shock protein 72 (HSP72) as a molecular chaperone has been confirmed to overexpress in epithelial carcinoma cells. There may be a possible association between the expression of HSP72 and HBx during the growth and progression of hepatocellular carcinoma cells. The aim of the study was to investigate the relationship between heat shock protein 72 and HBx in human hepatocellular carcinomas. The localization of HSP72 and HBx in human hepatocellular carcinomas was determined by immunohistochemistry and confocal laser microscopy. The interaction between HSP72 and HBx in liver cancer cells was analyzed by immunoprecipitation and Western blot. Our results revealed that hepatocellular carcinoma synchronously co-expressed higher level of HSP72 and HBx than that in the adjacent tissues to hepatocellular carcinoma. HSP72 and HBx were mainly immunolocalized in the cytoplasm. On the basis of immunoprecipitation and Western blot, we found that HSP72 formed complex with HBx in human hepatocellular carcinoma cells. The association between HSP72 and HBx in human hepatocellular carcinoma cells may contribute to study the pathogenesis and immunotherapy of hepatocellular carcinoma.

Rong G, Zhou Y, Xiong Y, Zhou L, Geng H, Jiang T, Zhu Y, Lu H, Zhang S, Wang P, Zhang B, Zhong R. · Department of Laboratory Medicine, Changzheng Hospital, Second Military Medical University, Shanghai, China. · Clin Exp Immunol. · Pubmed #19302244 No free full text.

Abstract: Primary biliary cirrhosis (PBC) is an organ-specific autoimmune liver disease characterized by progressive loss of intrahepatic small bile ducts. Cellular immune mechanisms involving T cell reaction are thought to be involved significantly in the pathogenesis of PBC. Recent studies have independently revealed enhanced T helper type 17 (Th17) response and weakened T regulatory cell (T(reg)) response in some autoimmune diseases, indicating a role of Th17/T(reg) imbalance in the pathogenesis of autoimmunity. This prompted us to investigate whether the Th17/T(reg) balance was broken in the peripheral blood of patients with PBC and, if it was, what cytokine circumstances might contribute to this imbalance. The expression of 11 Th17/T(reg) differentiation-related genes and serum concentrations of the corresponding cytokines in 36 patients with PBC, 28 patients with chronic hepatitis B and 28 healthy controls were measured by real-time quantitative-polymerase chain reaction and enzyme-linked immunosorbent assay respectively. Peripheral Th17 and T(reg) cells were analysed by flow cytometry. Th17-related cytokines were increased significantly in patients with PBC. Consistent with the cytokine profile, the Th17 cell population and retinoid-related orphan receptor gammat expression were increased markedly. In contrast, the T(reg) cell population and forkhead box P3 expression were decreased dramatically in the peripheral blood of patients with PBC. Our study revealed that the Th17/T(reg) imbalance, both cytokine profile and cell numbers, exists in patients with PBC, suggesting its potential role in the breakdown of immune self-tolerance in PBC. Interleukin-23, which characterized the imbalanced cytokine profile, may play an essential role in Th17-related human autoimmunity.

Wang LJ, Zhou Y, Hu YQ, Chang ZR. · Center for Disease Prevention and Control of Chongwen District, Beijing, China. · Zhonghua Yu Fang Yi Xue Za Zhi. · Pubmed #19115624 No free full text.

Abstract: OBJECTIVE: To understand the epidemiological features of viral hepatitis, and provide scientific evidence for developing strategies for prevention and control. METHODS: Through descriptive epidemiological methodology, the epidemiological characteristics of 1121 viral hepatitis cases in Chongwen District (1997-2006) were studied. chi2-test was used in ratio comparison. RESULTS: There were 1121 viral hepatitis cases occurring in the past decade, and the incidence rate was declining by year with the average of 27.10/ 100,000. Of which, the incidence rate of viral hepatitis B was the highest (13.90/ 100,000), viral hepatitis C (1.38/100,000) was the lowest. The proportion of viral hepatitis B was the highest (575, 51.29%) and viral hepatitis C (57, 5.08%) was the lowest. The incidence of viral hepatitis A, B and no-typing was declining year by year, while viral hepatitis C and E was climbing. Most viral hepatitis A and E cases occurred in Spring and Winter, and the difference of incidence rate between male and female was observed (chi2 = 188.39, P < 0.001). The findings also showed that viral hepatitis might occur in all kind of occupation, but most were workers (306, 27.3%) and officers (209, 18.64%). For the age distribution, 20 -49 years old group took the majority (749, 66.82%); and for the transmission route, blood transfusion (7.64/ 100,000) was significantly higher than fecal-oral route (3.02/ 100,000) (chi2 = 5.09, P < 0.01). CONCLUSION: It is necessary to raise and increase the public awareness for viral hepatitis prevention and control, and enhance immunization. Moreover, effective measures should be taken to the safety of blood and blood products, and to prevent nosocomial infection.

Ruebsam F, Tran CV, Li LS, Kim SH, Xiang AX, Zhou Y, Blazel JK, Sun Z, Dragovich PS, Zhao J, McGuire HM, Murphy DE, Tran MT, Stankovic N, Ellis DA, Gobbi A, Showalter RE, Webber SE, Shah AM, Tsan M, Patel RA, Lebrun LA, Hou HJ, Kamran R, Sergeeva MV, Bartkowski DM, Nolan TG, Norris DA, Kirkovsky L. · Department of Medicinal Chemistry, Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #19054673 No free full text.

Abstract: 5,6-Dihydro-1H-pyridin-2-one analogs were discovered as a novel class of inhibitors of genotype 1 HCV NS5B polymerase. Among these, compound 4ad displayed potent inhibitory activities in biochemical and replicon assays (IC(50) (1b)<10nM; IC(50) (1a)<25nM, EC(50) (1b)=16nM), good in vitro DMPK properties, as well as moderate oral bioavailability in monkeys (F=24%).

Bian T, Zhou Y, Bi S, Tan W, Wang Y. · National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Yingxin Street 100, Xuanwu Distract, Beijing 100052, PR China. · Biochem Biophys Res Commun. · Pubmed #19013428 No free full text.

Abstract: Although significant advances have been made on the studies of HCV glycoproteins (E1 and E2) recently, the role of the peptides preceding each glycoprotein remains unclear. We expressed E1 and E2 using two individual plasmids to form HCV pseudoparticles (HCVpp) in order to characterize the peptides preceding E1 and E2. Our data show that 14 amino acids from the HCV core and 12 amino acids from the E1 C-terminus are required for E1 and E2 function, respectively. The lack of a long enough peptide preceding E1 or E2 will abolish HCVpp infectivity, and the presence of fewer than 14 amino acids ahead of E1 and 12 amino acids ahead of E2 may alter their glycosylation. Furthermore, the peptides preceding E1 and E2 may be interchanged or may be replaced by those from genotype 2a. Our findings may contribute to the future development of new anti-HCV drugs.

Chen J, Wang Z, Guo Y, Peng J, Sun J, Ahmed CS, Zhou Y, Hou J. · Hepatology Unit and Department of Infectious Diseases, Nanfang Hospital, Nanfang Medical University, Guangzhou 510515, China. · Antiviral Res. · Pubmed #18948141 No free full text.

Abstract: Hepatitis B surface antigen (HBsAg) loss under antiviral therapy is rare in chronic hepatitis B patients and the dynamics of serum HBsAg in these patients are not available. The changes in serum HBsAg following treatment with adefovir (n=31) or peg-interferon-alpha-2a (n=23) were studied in hepatitis B e-antigen (HBeAg) positive chronic hepatitis B patients. Abbott Architect HBsAg assay was used to quantify serum HBsAg. HBsAg levels were significantly decreased during the first 12 weeks of treatment with median change of -397.0 IU/ml and -555.4 IU/ml, respectively for adefovir and peg-interferon-alpha-2a (p=0.005 and 0.001, respectively). Beyond 12 weeks, no further significant HBsAg reductions were found even in patients with sustained viral replication inhibition in either group. Three distinct patterns of HBsAg changes were observed in most patients in both treatment groups: biphasic pattern (rapid HBsAg reduction from baseline to week 12); assurgent pattern (higher HBsAg level at week 12 than at baseline); and wavy pattern (HBsAg reduction from baseline to week 12, followed by relapse at week 24 or week 28). These results might offer insights into the possible mechanism(s) underlying the unusual occurrences of HBsAg loss under antiviral therapy.

Du J, Zhou Y, Fu QX, Gong WL, Zhao F, Peng JC, Zhan LS. · Laboratory of Blood-borne Virus, Beijing Institute of Transfusion Medicine, Tai Ping Road 27, Beijing 100850, China. · FEBS Lett. · Pubmed #18822287 No free full text.

Abstract: By bioluminescence imaging and hydrodynamic gene transfer technology, the activities of hepatitis B virus (HBV) promoters and the effects of HBV enhancers on these promoters in mice under true physiological conditions have been assessed. Our studies reveal that either of the two HBV enhancers can stimulate HBV major promoter activity in hepa 1-6 cells (in vitro) and in mouse liver (in vivo), and the enhancer effects on the three promoters (S1, S2 and X promoter) are markedly greater in vivo than in vitro. The two HBV enhancers have no cooperative action on HBV promoters in vitro or in vivo.

Kim E, Li K, Lieu C, Tong S, Kawai S, Fukutomi T, Zhou Y, Wands J, Li J. · Liver Research Center, Rhode Island Hospital and Warren Alpert Medical School of Brown University, Providence, RI 02903, USA. · J Hepatol. · Pubmed #18809223 No free full text.

Abstract: BACKGROUND/AIMS: We previously reported that hepatitis C virus (HCV) core protein up regulated transcription of apolipoprotein C-IV (ApoC-IV, 10.7-fold increase), a member of the apolipoprotein family implicated in liver steatosis. Here, we identified host transcription factors regulating the ApoC-IV gene expression. METHODS: Transcriptional regulators were identified by DNA affinity purification and steatosis was detected by oil red O staining and triglyceride assay. RESULTS: We defined a 163-bp ApoC-IV promoter as a core protein responsive element, and identified Ku antigen complex (Ku70 and Ku80) as well as nuclear receptors PPARgamma/RXRalpha as key regulators of ApoC-IV gene expression. Both Ku70 overexpression and PPARgamma agonist significantly increased ApoC-IV promoter activity; conversely, Ku70 silencing or mutation of PPARgamma binding site diminished the ApoC-IV promoter activity. Interestingly, transient transfection of ApoC-IV cDNA into a human hepatoma cell line was able to trigger moderate lipid accumulation. In agreement with this in vitro study, ApoC-IV transcript level was increased in HCV infected livers which correlated with triglyceride accumulation. CONCLUSIONS: ApoC-IV overexpression may perturb lipid metabolism leading to lipid accumulation. HCV core protein may modulate ApoC-IV expression through Ku antigen and PPARgamma/RXRalpha complex.

Han M, Yan W, Guo W, Xi D, Zhou Y, Li W, Gao S, Liu M, Levy G, Luo X, Ning Q. · Department of Infectious Disease, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. · J Biol Chem. · Pubmed #18801734 No free full text.

Abstract: Fibrinogen-like protein 2 (FGL2)/fibroleukin plays a pivotal role in the pathogenesis of experimental and human fulminant and chronic viral hepatitis. To define the transcription factor(s) and upstream signal transduction pathways involved in the transcription of human FGL2 (hFGL2) in response to hepatitis B (HB) virus, hepatitis B core (HBc), hepatitis B virus S protein (HBs), or hepatitis B virus X protein (HBx) protein, expression plasmids were cotransfected with an hFGL2 promoter luciferase reporter construct into Chinese hamster ovary and HepG2 cells, respectively. HBc and HBx proteins, but not HBs protein, enhanced hFGL2 transcription in both cell lines. A strong regulatory region from -712 to -568 (relative to the transcriptional starting site) was shown to be responsible for hFGL2 gene transcription in response to both HBc and HBx proteins. c-Ets-2 was shown to be translocated to the nucleus in association with hFGL2 expression in response to both HBc and HBx proteins. Short hairpin RNA (shRNA) interference of c-Ets-2 expression inhibited hFGL2 gene transcription by 64.8 and 60.0% in response to HBc and HBx, respectively. c-Ets-2 protein was highly expressed in peripheral blood mononuclear cells from patients with severe chronic hepatitis B (CHB) in contrast to patients with mild CHB. Increased phosphorylation of ERK and JNK was detected in peripheral blood mononuclear cells from patients with severe CHB. ERK inhibitor PD098059 or ERK shRNA abolished the nuclear c-Ets-2 DNA binding activity and hFGL2 induction in response to HBc, whereas JNK inhibitor SP600125 or JNK shRNA abolished the nuclear c-Ets-2 DNA binding activity and hFGL2 induction in response to HBx. In conclusion, HBc and HBx proteins enhance transcription of hFGL2 through c-Ets-2 dependent on MAPK signal pathways.

Dragovich PS, Blazel JK, Ellis DA, Han Q, Kamran R, Kissinger CR, LeBrun LA, Li LS, Murphy DE, Noble M, Patel RA, Ruebsam F, Sergeeva MV, Shah AM, Showalter RE, Tran CV, Tsan M, Webber SE, Kirkovsky L, Zhou Y. · Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #18796353 No free full text.

Abstract: The synthesis of 4-(1',1'-dioxo-1',4'-dihydro-1'lambda(6)-benzo[1',2',4']thiadiazin-3'-yl)-5-hydroxy-2H-pyridazin-3-ones bearing 6-amino substituents as potent inhibitors of the HCV RNA-dependent RNA polymerase (NS5B) is described. Several of these agents also display potent antiviral activity in cell culture experiments (EC(50)<0.10 microM). In vitro DMPK data (microsome t(1/2), Caco-2 P(app)) for many of the compounds are also disclosed, and a crystal structure of a representative inhibitor complexed with the NS5B protein is discussed.

Ruebsam F, Sun Z, Ayida BK, Webber SE, Zhou Y, Zhao Q, Kissinger CR, Showalter RE, Shah AM, Tsan M, Patel R, Lebrun LA, Kamran R, Sergeeva MV, Bartkowski DM, Nolan TG, Norris DA, Kirkovsky L. · Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #18722768 No free full text.

Abstract: Hexahydro-pyrrolo- and hexahydro-1H-pyrido[1,2-b]pyridazin-2-one analogs were discovered as a novel class of inhibitors of genotype 1 HCV NS5B polymerase. Among these, compound 4c displayed potent inhibitory activities in biochemical and replicon assays (IC(50) (1b) <10 nM; EC(50) (1b)=34 nM) as well as good stability towards human liver microsomes (HLM t(1/2) =59 min).

Xiao W, Du L, Liang C, Guan J, Jiang S, Lustigman S, He Y, Zhou Y. · State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology & Epidemiology, Beijing 100071, China. · Vaccine. · Pubmed #18675867 links to  free full text

Abstract: Alum, the only adjuvant approved for clinical applications, can induce strong humoral (Th2) but weak cellular (Th1) immune responses. It is necessary to develop safe and effective adjuvants capable of inducing both humoral and cellular immune responses. We previously showed that activation-associated protein-1 (ASP-1) derived from Onchocerca volvulus has potent adjuvant activity. In this study, we have further evaluated the adjuvanticity of recombinant ASP-1 using a panel of recombinant proteins or synthetic peptide-based antigens, including ovalbumin (OVA), synthetic HIV peptide (HIV-p), recombinant HIV gp41 (rgp41) and HBV HBsAg, as well as three commercially available inactivated vaccines against haemorrhagic fever with renal syndrome (HFRS), Influenza and Rabies. Our results indicate that ASP-1 induced significantly higher IgG1 (Th2-associated) and IgG2a (Th1-associated) responses than alum adjuvant against OVA antigen, HIV-p, and rgp41. Consistently, it induced similar level of IgG1 responses as alum but higher level of IgG2a and IFN-gamma-producing T cell responses than alum adjuvant against HBsAg. Further, ASP-1 improved both IgG1 and IgG2a responses to three commercial inactivated vaccines when used separately or in combination. In conclusion, the recombinant ASP-1, unlike alum adjuvant, is able to induce both Th1 and Th2-associated humoral responses and Th1 cellular responses, suggesting that it can be further developed as a promising adjuvant for subunit-based and inactivated vaccines.

Yu Z, Wang Z, Chen J, Li H, Lin Z, Zhang F, Zhou Y, Hou J. · Department of Infectious Diseases, Nanfang Hospital, Nanfang Medical University, Guangzhou, China. · Arch Virol. · Pubmed #18668195 No free full text.

Abstract: Multiple studies have established that GTPase activity is critical for MxA to act against RNA viruses. Recently, it was shown that MxA can also restrict the replication of hepatitis B virus (HBV), a DNA virus, but the requirements for GTPase activity in inhibition of HBV by MxA remain unknown. Here, we report that GTPase-defective mutants (K83A, T103A, and L612K) can downregulate extracellular HBsAg and HBeAg and reduce the expression of extra- and intracellular HBV DNA in HepG2 cells to levels similar to that achieved by wild-type MxA. Furthermore, TMxA and T103, two nuclear forms of wild-type MxA and a GTPase-defective mutant (T103A) could only slightly decrease the expression of extra- and intracellular HBV DNA in HepG2 cells. In conclusion, GTPase activity is not essential for MxA protein to inhibit HBV replication, and MxA may have only a minimal effect on the replicative cycle of HBV in the nucleus.

Ellis DA, Blazel JK, Webber SE, Tran CV, Dragovich PS, Sun Z, Ruebsam F, McGuire HM, Xiang AX, Zhao J, Li LS, Zhou Y, Han Q, Kissinger CR, Showalter RE, Lardy M, Shah AM, Tsan M, Patel R, LeBrun LA, Kamran R, Bartkowski DM, Nolan TG, Norris DA, Sergeeva MV, Kirkovsky L. · Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #18662878 No free full text.

Abstract: 4-(1,1-Dioxo-1,4-dihydro-1lambda(6)-benzo[1,4]thiazin-3-yl)-5-hydroxy-2H-pyridazin-3-one analogs were discovered as a novel class of inhibitors of HCV NS5B polymerase. Structure-based design led to the identification of compound 3a that displayed potent inhibitory activities in biochemical and replicon assays (1b IC(50)<10 nM; 1b EC(50)=1.1 nM) as well as good stability toward human liver microsomes (HLM t(1/2)>60 min).

Bartels DJ, Zhou Y, Zhang EZ, Marcial M, Byrn RA, Pfeiffer T, Tigges AM, Adiwijaya BS, Lin C, Kwong AD, Kieffer TL. · Department of Infectious Diseases, Vertex Pharmaceuticals, Inc., Cambridge, Massachusetts 02139, USA. · J Infect Dis. · Pubmed #18637752 No free full text.

Abstract: BACKGROUND: The prevalence and clinical implications of naturally occurring variants that are resistant to hepatitis C virus (HCV) protease inhibitors in treatment-naive patients has not been reported. We report here the prevalence of such variants and their effect on clinical response. METHODS: Population sequence analysis of the NS3.4A protease was conducted in 570 treatment-naive subjects. RESULTS: Most subjects (98%) had wild-type virus. The remaining subjects had the following variants present in significant proportions (100%): V36M, 0.9%; R155K, 0.7%; V170A, 0.2%; and R109K, 0.2%. The V36M, R109K, and V170A substitutions confer low-level resistance (<7-fold) to protease inhibitors in replicon cells. The R155K substitution confers low-level resistance to telaprevir (TVR) and boceprevir and confers high-level resistance (>70-fold) to BILN 2061 and ITMN-191. Five subjects with the V36M or R109K variant were treated with 8-24 weeks of TVR and peginterferon-alpha2a (P) with or without ribavirin (R). Four achieved a sustained viral response, and 1 was lost to follow-up. In subjects with the R155K variant, TVR/PR provided greater antiviral activity than PR alone; however, the antiviral response was lower than that observed in subjects with wild-type virus. CONCLUSION: High levels of naturally occurring protease inhibitor-resistant variants were uncommon (<1% each) in HCV treatment-naive patients. TVR/PR efficiently inhibited V36M and R109K variants and contributed partial antiviral activity against the R155K variant. As new HCV agents are evaluated in clinical trials, it will be important to monitor the effect of baseline variants on sensitivity.

Zhou Y, Park CM, Cho CW, Song YS. · Department of Food and Nutrition, Food Science Institute and BPRC, Inje University, Obang-dong, Gimhae, Gyeongnam 621-749, Korea. · Biosci Biotechnol Biochem. · Pubmed #18603811 links to  free full text

Abstract: The protective effect of pinitol against D-galactosamine (GalN)-induced liver damage was examined. Forty male Sprague-Dawley rats were divided into normal control, GalN control, and pinitol groups (0.5%, 1%, and 2%). After 8 weeks of feeding, a single dose of GalN (650 mg/kg) was administered 24 h before their sacrifice. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and tumor necrosis factor-alpha (TNF-alpha) levels were significantly increased after an injection with GalN (P<0.05), but pinitol supplementation at the level of 0.5% reversed these changes to normal levels. Significant decreases in serum triglyceride and cholesterol and increases in hepatic cholesterol were observed in GalN-intoxicated rats. However, supplementation with pinitol significantly attenuated these trends. In addition, pinitol elevated the Mn-superoxide dismutase, glutathione reductase, and catalase activities, prevented hepatic lipid peroxidation, and restored the hepatic GSH levels and cytochrome P450 2E1 function. Thus, 0.5% pinitol supplementation protected the rats from the hepatotoxicity induced by GalN, at least part of its effect being attributable to attenuation of the oxidative stress and inflammatory process promoted by GalN.

Kim SH, Tran MT, Ruebsam F, Xiang AX, Ayida B, McGuire H, Ellis D, Blazel J, Tran CV, Murphy DE, Webber SE, Zhou Y, Shah AM, Tsan M, Showalter RE, Patel R, Gobbi A, LeBrun LA, Bartkowski DM, Nolan TG, Norris DA, Sergeeva MV, Kirkovsky L, Zhao Q, Han Q, Kissinger CR. · Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #18554907 No free full text.

Abstract: A novel series of HCV NS5B polymerase inhibitors comprising 1,1-dioxoisothiazoles and benzo[b]thiophene-1,1-dioxides were designed, synthesized, and evaluated. SAR studies guided by structure-based design led to the identification of a number of potent NS5B inhibitors with nanomolar IC(50) values. The most potent compound exhibited IC(50) less than 10nM against the genotype 1b HCV polymerase and EC(50) of 70 nM against a genotype 1b replicon in cell culture. The DMPK properties of selected compounds were also evaluated.

Luo K, Mao Q, Karayiannis P, Liu D, Liu Z, Zhou Y, Feng X, Zhu Y, Guo Y, Jiang R, Zhou F, Peng J, Hou J. · Department of Infectious Diseases, Nanfang Hospital, Guangzhou, China. · J Viral Hepat. · Pubmed #18554243 No free full text.

Abstract: The response to interferon-alpha treatment of patients with chronic hepatitis B under the current protocol is not satisfactory. The aim of this study was to try an alternative approach to improve treatment outcome. Of 374 HBeAg-positive patients, 127 of them received 5 million units of interferon-alpha thrice weekly for 6 months and constituted the control group, while 247 in the study group received the same dosage but the duration of treatment was tailored. The study protocol provided for continuation of treatment if HBV DNA levels were continuously decreasing. The treatment ended when viral, antigenic and biochemical endpoints were reached or when HBV DNA levels were no longer decreasing. The median length of tailored treatment was 10 (range 6-24) months. The end-of-treatment response rates were 39.3% and 23.6% (P = 0.002), and after 12-month, follow-up, the sustained response rates were 40.5% and 28.3% (P = 0.013) in the study and control groups, respectively. Excluding the patients who dropped out, 228 and 115 completed a median of 40- and 44-month-long follow-up; the long-term response was thus 45.3% and 33.1% (P = 0.014) in the respective groups. Interferon-alpha treatment tailored in length demonstrated significantly increased efficacy in patients with chronic hepatitis B.

Wu X, Zhou Y, Zhang K, Liu Q, Guo D. · State Key Laboratory of Virology, the Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, PR China. · FEBS Lett. · Pubmed #18519040 No free full text.

Abstract: Hepatitis C virus (HCV) replication and pathogenesis involve both virus-encoded proteins and cellular factors. In our study, we showed that NS5B, the HCV RNA-dependent RNA polymerase, interacted with M2 type pyruvate kinase (M2PK) but not L type pyruvate kinase. We confirmed the interaction by GST pull down, coimmunoprecipitation and confocal immunofluorescence analysis in cells with transient expression of NS5B and M2PK as well as in a HCV replicon-bearing cell line. Furthermore shRNA which specifically down-regulated M2PK expression could inhibit the replication of HCV in HCV replicon 9B cells.

Ruebsam F, Webber SE, Tran MT, Tran CV, Murphy DE, Zhao J, Dragovich PS, Kim SH, Li LS, Zhou Y, Han Q, Kissinger CR, Showalter RE, Lardy M, Shah AM, Tsan M, Patel R, Lebrun LA, Kamran R, Sergeeva MV, Bartkowski DM, Nolan TG, Norris DA, Kirkovsky L. · Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #18487044 No free full text.

Abstract: Pyrrolo[1,2-b]pyridazin-2-one analogs were discovered as a novel class of inhibitors of genotype 1 HCV NS5B polymerase. Structure-based design led to the discovery of compound 3 k, which displayed potent inhibitory activities in biochemical and replicon assays (IC(50) (1b)<10nM; EC(50) (1b)=12 nM) as well as good stability towards human liver microsomes (HLM t(1/2)>60 min).

Peng JS, Zhou DJ, Pei DE, Zhou Y, Liu MQ, Tang L, Xu J, Wu XW, Huo WZ, Zhou W. · Tongji Medical College, Hua Zhong University of Science & Technology, Wuhan 430030, China. · Zhonghua Liu Xing Bing Xue Za Zhi. · Pubmed #18476583 No free full text.

Abstract: OBJECTIVE: To study the prevalence of hepatitis C virus (HCV) infection and characteristics on molecular biology related to HCV among patients who were enrolled in a Methadone maintenance clinic in Wuhan. METHODS: Serum samples from 332 injection drug users (IDUs) were obtained and anti-HCV IgG was detected by enzyme linked immunosorbrent assay(ELISA), together with 86 anti-HCV positive specimens genotyped. A reverse transcriptase-polymerase chain reaction (RT-nPCR) assay using conserved primers deduced from the core-envelopel (C-E1) region of the HCV genome was employed to amplify a 474 bp fragment. Phylogenetic analysis of the C-E1 sequences was conducted by direct sequencing of the RT-nPCR products and alignment with determined by nucleotide sequencing followed by composition of a phylogenetic tree. RESULTS: There were 313 cases (94.3%) appeared positive anti-HCV IgG in the 332 patients from a Methadone maintenance clinic in Wuhan. It was demonstrated that there were four different subtypes of HCV in that clinic in Wuhan, including 6a--71 cases (82.5%), 3b--7 cases (8.2%), 1a--5 cases (5.8%) and 1b--3 cases (3.5%). CONCLUSION: Infection of 6a genotype HCV was predominant in patients from the Methadone maintenance clinic in Wuhan, followed by HCV 3b, 1a and 1b.

Li LS, Zhou Y, Murphy DE, Stankovic N, Zhao J, Dragovich PS, Bertolini T, Sun Z, Ayida B, Tran CV, Ruebsam F, Webber SE, Shah AM, Tsan M, Showalter RE, Patel R, Lebrun LA, Bartkowski DM, Nolan TG, Norris DA, Kamran R, Brooks J, Sergeeva MV, Kirkovsky L, Zhao Q, Kissinger CR. · Anadys Pharmaceuticals, Inc., 3115 Merryfield Row, San Diego, CA 92121, USA. · Bioorg Med Chem Lett. · Pubmed #18457949 No free full text.

Abstract: 5-Hydroxy-3(2H)-pyridazinone derivatives were investigated as inhibitors of genotype 1 HCV NS5B polymerase. Lead optimization led to the discovery of compound 3a, which displayed potent inhibitory activities in biochemical and replicon assays [IC(50) (1b)<10nM; IC(50) (1a)=22 nM; EC(50) (1b)=5nM], good stability toward human liver microsomes (HLM t(1/2)>60 min), and high ratios of liver to plasma concentrations 12h after a single oral administration to rats.