Association of transforming growth factor-beta1 gene polymorphisms with a hepatocellular carcinoma risk in patients with chronic hepatitis B virus infection

Transforming growth factor-beta1 (TGF-beta1) can act as both a tumor suppressor and a stimulator of tumor progression. We have examined the relationship between polymorphisms of the TGF-beta1 gene and the risk of hepatocellular carcinoma (HCC) in patients with chronic hepatitis B virus (HBV) infection. A total of 1,237 Korean subjects were prospectively enrolled; 1,046 patients with chronic HBV infection and 191 healthy controls with no evidence of recent or remote HBV infection. The patients were divided into two groups: those without (n = 809) and those with HCC (n = 237). Single nucleotide polymorphisms (SNPs) of TGF-beta1 were searched for and genotyped using the single base extension method. In Korean subjects, only two SNPs were found among the seven known polymorphisms of TGF-beta1, at position -509 and in codon 10. The risk of HCC was significantly lower in patients with the T/T or C/T genotypes than in those with the C/C genotypes at position -509 (P < 0.02), and also lower among those with the Pro/Pro or Leu/Pro genotypes than in those with the Leu/Leu genotypes in codon 10 (P < 0.007). Haplotype analysis revealed that the possession of [-509C > T; L10P] conferred a decreased likelihood of HCC (OR = 0.74; 95% CI, 0.59-0.93; P = 0.008). In conclusion, the presence of the TGF-beta1 -509C > T promoter or of the L10P polymorphism, and the combination of both [-509C > T; L10P] as a haplotype were strongly associated with a reduced risk of HCC in patients with chronic HBV infection.     

Computational efforts to identify natural occurring compounds from phyllanthus niruri that target hepatitis B viral infections: DFT,docking and dynamics simulation study

Hepatitis B virus (HBV) is an enveloped hepatotropic virus responsible for nucleic acids replication. It causes chronic infection. Depending on the strain, mutations in the core protein of chronic hepatitis B virus (HBV) infections occur. Medicinal plants, the backbone of traditional medicine, are a potential source of lead molecules in drug discovery due to extensive pharmacological studies. In this study, we have screened twenty-nine phytochemicals. The ADME and drug-likeness of these phytochemicals were investigated. After screening, the binding affinity of ten phytochemicals was studied through molecular docking. Simulation studies were carried out for 100 ns to analyze the properties of RMSD, Rg, RMSF, average hydrogen bond number and SASA of hepatitis B virus capsid protein. As per the docking results phyllanthosterol, may be used as a potential inhibitors against HBV. The simulations findings revealed that, in case of mutant protein, the flexibility nature decreases as compared to wildtype protein. Our results may provide useful information for drug design and to lead the identification of novel inhibitor for hepatotropic viral infection.     

Bioinspired Artificial Nanodecoys for Hepatitis B Virus

A facile route is presented for fabricating a new class of nanomimics that overexpress hepatitis B virus (HBV) receptor by a natural biosynthetic procedure against HBV infection. A nine‐transmembrane HBV‐specific receptor, human sodium taurocholate co‐transporting polypeptide (hNTCP), was engineered to naturally immobilize it onto the cellular surface and subsequently trigger the budding of hNTCP‐anchoring membrane vesicles (hNTCP‐MVs) that favor the HBV virion. hNTCP‐MVs could rapidly block HBV infection in cell models. Furthermore, hNTCP‐MVs treatment could effectively prevent viral infection, spreading, and replication in a human‐liver‐chimeric mouse model of HBV infection. Our findings demonstrate the receptor‐mediated antiviral effect of hNTCP‐MVs to trick HBV and offer novel opportunities for further development of antiviral strategies in nanomedicine.     

Synthesis of Novel Nucleoside: 2'-deoxyuridine-3',5'-di(4, 4'-dimethoxy-5, 6, 5', 6'-dimethylenedioxybipheny1-2-methoxy carbonyl 2'-carboxylate)

Anumberofnucleosideshavebeenreportedtobeanti-hepatitisBvirusagents',althoughnoneofthemhaveyetbeenprovedtobeclinicallyusefull.Inaddition,dimethyldicarboxylatebiphenyl(DDB)isawidelyusedanti-hepatiticdrugwithhigheffectinloweringSGPTandconsiderablelowtoxicity'.InviewofthediscoverythatseveralnucleosideswiththeD-configurationandanaloguesofthebiphenyldicarboxylateareselectiveantiviralagents,itwasofinteresttosynthesizenucleosidederivativesaspotentialantiviralagents.Recently,thecarbocyclicanalogueo…     

LC-MS characterization of efficacy substances in serum of experimental animals treated with Sophora flavescens extracts

Anti-DHBV (duck hepatitis B virus) activity was found in the aqueous extracts of Sophora flavescens Ait. in vivo. Liquid chromatography/electrospray ionization ion trap mass spectrometry was applied to characterize the components in duck serum after oral administration of S. flavescens extract. Oxymatrine (1), sophoranol (2), sophoridine (3) and matrine (4) were identified in the serum. Further research on the four compounds was evaluated for their antiviral activity against HBV (hepatitis B virus) in cell culture. The results suggested that oxymatrine, sophoranol and matrine were the efficacy substances for anti-HBV activity in aqueous extracts of S. flavescens Ait.     

Mutation detection in the drug-resistant hepatitis B virus polymerase gene using nanostructured reverse micelles.

The emergence of drug-resistant hepatitis B virus (HBV) has been reported in patients with prolonged administration of lamivudine, which is a potent drug for the prevention of HBV infection. Lamivudine-resistant HBV has several types of mutations at the YMDD motif of its DNA polymerase. We successfully demonstrated that monitoring the hybridization behavior in nanostructured reverse micelles enables us to detect single nucleotide polymorphisms (SNPs). With the aid of reverse micelles, a model 40-mer oligonucleotide containing a single-base substitution was clearly distinguished from the normal, complementary oligonucleotide. In addition, we extended this technique to a high-throughput analysis. The results obtained with a 96-well micro-plate reader indicated the possibility of SNPs detection toward multiple samples of patients.     

Current Progress in the Development of Hepatitis B Virus Capsid Assembly Modulators: Chemical Structure,Mode-of-Action and Efficacy

Hepatitis B virus (HBV) is a major causative agent of human hepatitis. Its viral genome comprises partially double-stranded DNA, which is complexed with viral polymerase within an icosahedral capsid consisting of a dimeric core protein. Here, we describe the effects of capsid assembly modulators (CAMs) on the geometric or kinetic disruption of capsid construction and the virus life cycle. We highlight classical, early-generation CAMs such as heteroaryldihydropyrimidines, phenylpropenamides or sulfamoylbenzamides, and focus on the chemical structure and antiviral efficacy of recently identified non-classical CAMs, which consist of carboxamides, aryl ureas, bithiazoles, hydrazones, benzylpyridazinones, pyrimidines, quinolines, dyes, and antimicrobial compounds. We summarize the therapeutic efficacy of four representative classical compounds with data from clinical phase 1 studies in chronic HBV patients. Most of these compounds are in phase 2 trials, either as monotherapy or in combination with approved nucleos(t)ides drugs or other immunostimulatory molecules. As followers of the early CAMs, the therapeutic efficacy of several non-classical CAMs has been evaluated in humanized mouse models of HBV infection. It is expected that these next-generation HBV CAMs will be promising candidates for a series of extended human clinical trials.     

Association between interleukin 6 promoter variants and chronic hepatitis B progression

Interleukin 6 (IL6) plays an essential role in the regulation of immune response to chronic disease. In this study, the three known single nucleotide polymorphisms (SNPs) in the IL6 promoter region were genotyped in a large chronic hepatitis B cohort to evaluate the effects of IL6 promoter variants. The single base extension method was used for this genotyping. Haplotypes were constructed by the three SNPs in IL6. Allele frequencies were compared for; i) patients with chronic hepatitis (CH) and chronic carriers vs. chronic hepatis patients with clinical evidence of liver cirrhosis (LC) (i.e., portal hypertension), ii) cirrhotic patients with hepatocellular carcinoma (HCC) vs. without HCC by logistic regression, and iii) with respect to the time intervals from the onset of infection to HCC. Results were analyzed by Cox relative hazard analysis on the assumption that all the patients were infected during early infancy. The frequencies of each SNP were 0.002 (IL6-597 G>A), 0.25 (IL6-572 C>G) and 0.002 (IL6-174 G>C), respectively, in the Korean population (n = 1,046). No significant associations were detected between IL6-572 C>G and chronic hepatitis B outcome in this study; i.e., LC occurrence on CH (OR = 0.16-1.27, P = 0.13- 0.71) and HCC occurrence on LC (OR = 1.04-1.23, P = 0.89-0.60) of heterozygotes and homozygotes for G allele in referent comparison to homozygotes for common allele (C/C genotype), and time interval to HCC (RH = 0.67-1.00; P = 0.14-0.99). In conclusion, there appeared to be no significant associations between IL6 promoter variants and disease outcome in chronic hepatitis B.     

Medicinal chemistry strategies in the discovery and optimization of HBV core protein allosteric modulators (2018–2022 update)

Despite the improving coverage of preventative vaccines, hepatitis B remains a severe global public health problem, with more than 250 million patients living with hepatitis B virus (HBV) infection. Current available therapies, including nucleos(t)ide analogs and peginterferon, can control HBV replication but fail to eliminate covalently closed circular DNA (cccDNA) and achieve a cure. The HBV core protein (Cp) is a well-conserved structural protein, self-assembling to form the viral capsid. It involves in or modulates almost every stage of the HBV lifecycle, which makes it an attractive target for the development of new anti-HBV therapies. HBV core protein allosteric modulators (CpAMs) have become a hotspot in recent years. Herein, we provide a concise report focusing on the various medicinal chemistry strategies involved in the latest research (2018–2022) of HBV CpAMs, including high throughput screening (HTS), virtual screening (VS), drug repositioning, natural products, substitution decorating approach, scaffold hopping, molecular hybridization, prodrug strategy and conformational constraint strategy, to provide guidance for further development of new and effective anti-HBV drugs.     

The potential of magnetic nanocluster and dual-functional protein-based strategy for noninvasive detection of HBV surface antibodies

Magnetic nanoclusters (MNCs) were synthesized in a one-pot process, carboxylic MNCs and dual-functional protein were prepared and used to capture hepatitis B virus surface antibodies (anti-HBs) in simulated diseased oral mucosal transudate (OMT) samples. The specific substrate of dual-functional protein, dual-labeled double-chained DNA molecules, based on Fluorescence Resonance Energy Transfer (FRET), was used to amplify the detection signal and the detection limit of 0.1 ng mL(-1) of anti-HBs monoclonal antibodies was achieved. Combination MNCs with dual-functional protein enables the noninvasive detection of hepatitis B virus (HBV) surface antibodies in OMT samples, showing promise as a diagnostic tool for the OMT diagnosis of infectious diseases with sensitive, specific and facile capabilities.     

纳米探针芯片技术用于微量乙肝病毒DNA的检测

利用两组探针修饰的微粒:(1)表面标记有可与待测乙肝病毒(HBV) DNA另一端结合的纳米金探针1(信号探针)以及可与信号探针部分结合的纳米金探针2(检测探针);(2)表面标记有可与待测HBV DNA一端结合的磁珠探针(捕捉探针1).检测靶HBV DNA时,磁珠探针与信号探针在液相中可分别与HBV DNA靶序列一端结合最终形成三明治样结构.再以磁场将三明治样复合物从反应液中分离,以DTT溶液将信号探针从纳米金颗粒上洗脱.洗脱后的信号探针数量反映靶基因的多寡,信号探针一段与预先点样的基因芯片上的捕捉探针2结合,检测探针与信号探针另一段相结合,最后用银染液将检测探针显色从而得到靶目标DNA相对定量信息.结果表明,本检测方法的检测灵敏度达到10-15 mol/L水平.检测时间少于1.5 h,检测结果与HBV DNA水平呈现较好的线性关系且无假阳性结果;本方法有望用于乙肝病人血清中HBV DNA的快速筛测及其它微生物基因的检测.     

Characterization of hepatitis B virus capsids by resistive-pulse sensing

We report characterization of hepatitis B virus (HBV) capsids by resistive-pulse sensing through single track-etched conical nanopores formed in poly(ethylene terephthalate) membranes. The pores were ～40 nm in diameter at the tip, and the pore surface was covalently modified with triethylene glycol to reduce surface charge density, minimize adsorption of the virus capsids, and suppress electroosmotic flow in the pore. The HBV capsids were assembled in vitro from Cp149, the assembly domain of HBV capsid protein. Assembled T = 3 (90 Cp149 dimer) and T = 4 (120 dimer) capsids are 31 and 36 nm in diameter, respectively, and were easily discriminated by monitoring the change in current as capsids passed through an electrically biased pore. The ratio of the number of T = 3 to T = 4 capsids transiting a pore did not reflect actual concentrations, but favored transport of smaller T = 3 capsids. These results combined with longer transit times for the T = 4 capsids indicated that the capsids must overcome an entropic barrier to enter a pore.     

Synthesis and anti-HBV activity of novel 5-substituted pyridin-2(1H)-one derivatives

<正>Four novel 5-substituted pyridine-2(1H)-one derivatives were designed and synthesized by using addition-elimination reactions.The structures of these novelly synthesized compounds were verified by ~1H NMR,ESI-MS and single crystal X-ray diffraction.Furthermore,all four compounds(most notably compound 7a) were found to be highly efficient against hepatitis B virus (HBV) in cultured HepG2 2.2.15 cell,making them promising drug candidates for potential bioactive molecule against hepatitis B.     

高效液相色谱法同时测定妊娠合并乙型肝炎患者血浆中的吲哚与3-甲基吲哚

建立了一种高效液相色谱-荧光检测法用于同时测定血浆中的吲哚与3-甲基吲哚。样本经液液萃取法提取,采用Shim-Pack VP-ODS柱(150 mm×4.6 mm,4.6μm),以15 mmol/L磷酸二氢钠溶液-甲醇(40∶60,v/v)为流动相,甲奈酚为内标,荧光激发和发射波长分别为274 nm和340 nm。吲哚和3-甲基吲哚的线性范围分别为2.22~88.89μg/L和1.11~44.44μg/L;检出限分别为0.11μg/L(吲哚)和0.06μg/L(3-甲基吲哚);平均回收率为95.5%~112.3%,日内与日间相对标准偏差均小于6.8%。利用该方法对妊娠合并乙肝患者(n=29)和正常孕妇(n=46)的血浆进行了测定,结果表明妊娠合并乙肝患者血浆中吲哚和3-甲基吲哚水平均显著高于正常对照组,且与肝损伤指标转氨酶水平呈正相关。     

Deoxy Derivatives of L-like 5'-Noraristeromycin

Several base variations of 2'- and 3'-deoxy derivatives of (+)-4'-deoxy-5'-noraristeromycin have been prepared from enantiomerically pure precursors following standard purine nucleoside construction. These carbocyclic nucleosides were evaluated against hepatitis B virus (HBV) and found to be inactive. No cytotoxicity to the cell line was observed.     

血液乙肝病毒基因检测

全世界每年死于乙型肝炎病毒相关疾病的人数已达60万人,应用荧光定量PCR法检测能够提高乙肝病毒基因的检出率,正确掌握该技术方法能够更加有效地进行疾病诊断和预防工作。基于此,研究针对荧光定量PCR法检测乙肝病毒基因的方法进行探讨。