Comprehensive Quality and Bioactive Constituent Analysis of Celery Juice Made from Different Cultivars.

Celery juice is rich in bioactive constituents, has good health properties, and is becoming much more popular, with its demand continuing to rise. The results of this study show that celery juice from Chinese cultivars contains more bioactive constituents, whereas celery cultivars from the United States and European countries have a higher juice yield. Compared with the other juices, the juices of five cultivars may taste sweeter, and the juices of three cultivars had a higher antioxidant capacity. The juices of six cultivars (three with the highest antioxidant capacity and three with the lowest antioxidant capacity) were selected to analyze bioactive constituents by LC/MS and GC/MS. A total of 71 phenolic acids, 38 flavonoids, 18 coumarins, 41 terpenoids, and 11 phthalides were detected in the juices of the six celery cultivars. The contents of 14 compounds had a more than 10-fold difference among these celery juices. This study first evaluated the comprehensive quality of the juices made from 26 celery cultivars and then analyzed the differences in bioactive constituents in the juices of6 celery cultivars. These findings provide information for the further study on the health functions of celery juice and can also guide celery juice production and celery breeding.

Evaluation of the safety, tolerability, and pharmacokinetics of RO7049389 in healthy Chinese volunteers.

The objectives of this phase I study are to assess the safety, tolerability, and pharmacokinetics (PKs) of RO7049389 in healthy Chinese volunteers (HVs) and evaluate potential ethnic differences in the safety and PKs using data from this study and the first-in-human study (in which most of the HVs were non-Asian). HVs randomly received a single dose of 200-600 mg of RO7049389 or a placebo in a single ascending dose (n = 28) or multiple doses of 200-400 mg of RO7049389 or a placebo in multiple ascending doses (n = 24). Safety and tolerability were monitored throughout the study. Serial blood samples were collected for PK analysis. RO7049389 was safe and well-tolerated in the HVs. The time to maximum concentration ranged from 1.5 to 3.0 h, and terminal half-life ranged from 3.66 to 14.6 h. A single dose of 200-600 mg and multiple doses of 200-400 mg exhibited nonlinear PKs. In general, the safety profiles were comparable between non-Asian and Asian HVs, but the plasma exposure of RO7049389 in Chinese HVs was higher than that in non-Asian HVs. The data generated from this study will provide guidance for future clinical studies on RO7049389 in Chinese/Asian patients with hepatitis B virus.

Further Evaluation of Coproporphyrins as Clinical Endogenous Markers for OATP1B.

Coproporphyrins (CP-I and CP-III) in plasma are considered potential markers for assessing liver organic anion-transporting polypeptide transporter OATP1B activity and monitoring OATP1B-mediated drug-drug interactions (DDIs) in clinical settings. However, the effect of altered renal clearance (CLrenal ) on CP-I and CP-III plasma exposure has rarely been examined. Therefore, the purpose of this study is to further evaluate CP-I and CP-III as clinical endogenous markers for OATP1B activity and to investigate the impact of CLrenal on DDI assessments for the first time. In this study, 18 healthy participants were recruited to receive RO7049389 (a potential inhibitor of OATP1B) 800 mg twice daily for 6 days and a single dose of pitavastatin (a probe drug of OATP1B) before and after RO7049389 treatment. Plasma concentrations of pitavastatin, CP I, CP III, and the amounts of CP-I and CP-III excreted in urine were measured. Seventeen healthy participants completed the study. After multiple doses of RO7049389, the area under the plasma concentration-time curve from time 0 to 12 hours of pitavastatin increased 1.95-fold (90% confidence interval [CI], 1.58-2.41), while for CP-I and CP-III it increased 3.00-fold (90%CI, 2.35-3.82) and 2.84-fold (90%CI, 2.22-3.65), respectively. Concurrently, the CLrenal of CP-I decreased by 31% (90%CI, 23%-39%), and that of CP-III decreased by 70% (90%CI, 61%-77%). In conclusion, CP-I and CP-III in plasma display the potential to be applied as endogenous markers for the evaluation of OATP1B inhibition in clinical trials. While renal transporters contribute significantly to the CLrenal of CP-III, it would be better to investigate the impact of the CLrenal on plasma exposure of CP-III during clinical DDI assessments.

A Five-in-One First-in-Human Study To Assess Safety, Tolerability, and Pharmacokinetics of RO7049389, an Inhibitor of Hepatitis B Virus Capsid Assembly, after Single and Multiple Ascending Doses in Healthy Participants.

RO7049389, an inhibitor of hepatitis B virus (HBV) capsid assembly, is being developed for the treatment of patients with chronic HBV infection. The objectives of this first-in-human study are to assess the safety, tolerability, pharmacokinetics (PK), food effect, inhibitory effect on CYP3A, and effect on QT of RO7049389 in healthy participants. Five components, single-ascending-dose (SAD) cohorts, multiple-ascending-dose (MAD) cohorts, food effect assessment, drug-drug interaction assessment, and concentration-QT analysis were integrated in one study (five-in-one). Participants randomly received a single dose of 150 to 2,500 mg RO7049389 or placebo in SAD cohorts (n = 41), or multiple doses of 200 to 800 mg RO7049389 or placebo in MAD cohorts (n = 42). A single doses of 450 mg RO7049389 was administered under fasted and fed condition. The microdose of midazolam was administered before and after multiple dosing of RO7049389. Safety and tolerability were monitored throughout the study. Serial blood and urine samples were collected for the PK analysis. RO7049389 was safe and well tolerated in healthy participants. Absorption and elimination of RO7049389 occurred rapidly in plasma with minimal recovery in urine. Greater than dose-proportional increases in plasma exposure were observed. Exposure of RO7049389 (450 mg) increased by ∼2-fold when administered with a high-fat meal. The inhibition effect of RO7049389 on CYP3A was weak (<20%). No effect on QT interval was observed at up to a single dose of 2,500 mg. RO7049389 displayed a favorable safety, tolerability and PK profile suitable for further clinical development. (This trial was registered at ClinicalTrials.gov with the identifier NCT02952924.).

Full-length hepatitis B virus sequences from naïve patients with fluctuation of viral load during ADV monotherapy.

The reasons for adefovir dipivoxil (ADV) treatment failures appear diverse. Few studies have reported full-length hepatitis B virus (HBV) genome in patients with ADV treatment failures. The patients were from a phase III clinical trial that investigated the antiviral response to ADV in China. Seven patients had increase in HBV-DNA (>1 log(10) copies/ml above on-treatment nadir) at week 52. The serum HBV-DNA levels were above 10(4)copies/ml at week 92 in four of them. Sixteen full-length HBV genomes from the four patients at four time points were sequenced using cloning sequencing method. The frequency of substitutions at week 52 was higher than at weeks 28(16 wt) and 92(80). HBV-DNA reduction was correlated negatively with the frequency of substitutions at the three time points. No published ADV-resistant mutations were detected. The mutations, including substitutions in immunogenic epitopes and conserved sites of the polymerase gene, were frequent during ADV treatment. Amino acid deletions in X gene and basal core promoter/pre-core mutations appeared before or during ADV treatment. The substitutions in immunogenic epitopes (mainly of the surface gene) and conserved sites of the polymerase gene other than ADV-resistant mutations may have influenced antiviral efficacy in the study. More potent antiviral drugs may be important to rescue individual patients and for public health safety. It is needed to study how these substitutions influence HBV replication, disease progression, and antiviral treatment efficacy.

[The change of HBV nucleotide sequence during Adefovir dipivoxil treatment].

OBJECTIVE: To study the change of HBV nucleotide sequence during Adefovir dipivoxil treatment. METHODS: Serum samples were collected from 4 patients (numbers 228, 229, 230 and 233) with chronic hepatitis B (CHB) treated with PMEA. PCR amplification of full-length genomes, total genome clone, and sequencing and linkage of the HBV viruses were done with the serum samples collected before the therapy, and again on the 28th week, 52nd week and on the 92nd week of the therapy. Gene types and serum types were studied according to the HBV DNA sequence. The related biological information was analyzed, including the homogeneity and heterogeneity of the HBV DNA sequence before and during PMEA therapy, and variance of amino acids coded by HBV sequence. RESULTS: The gene types and serum types of HBV of the 4 patients were determined. Patient 228 was gene type C/serum type adrq+. Patient 229 was complex gene type B and C/complex serum type adw2 and adrq+. Patients 230 and 233 were both serum type adw2. The homogeneity of total gene sequence was 97.5% - 99.8% in one group of patients and 90.6% - 100% between groups in variant patients. The base mutation quantity of 52W was significantly higher than that of 28W and 92W. There was a common site of hotspot congregating mutation that existed in the P region. The target of PMEA was located in the P region encoding reverse transcriptase, which was generally very conservative. Therefore the mutations in this region were very significant. This might affect the activity of reverse transcriptase, leading to resistance to PMEA. CONCLUSION: We discovered some unusual aminophenol variations in the HBV genomes, which most probably are related to the HBV mutation that resulted in the developing resistance to PMEA.

[SHP-1 gene's methylation status of Daudi lymphoma cell and the demethylation effect of 5-aza-2'-deoxycytidine].

OBJECTIVE: To explore the transcription regulation of 5-aza-2'-deoxycytidine(5-Aza-CdR) on SHP-1 gene and its effects on Daudi cell line growth. METHODS: MTT method and flow cytometry were used to detect the growth and apoptosis of Daudi cells after treated with different dosage of 5-Aza-CdIR. Bisulfite sequencing PCR ( BSP) , T-A cloning and sequence analysis were evaluated for methylation status. The SHP-I mRNA and protein were determined by reverse transcription polymerase chain reaction (RT-PCR) ,immunohistochemistry. RESULTS: (1)After 7 d treatment with 2. 00 micromol/L of 5-Aza-CdR, all cytosines (C) in Daudi cells genome DNA were converted to thymidine, and SHP-1 mRNA and protein expressed again in the cells while those Cs in CpG dinucleotides in untreated Daudi cells remained Cs; (2)5-Aza-CdR inhibited the cell growth,The effects within certain extent were dose and time dependent:after 72 h treatment with 5-Aza-CdR at 200. 00, 20. 00, 2. 00 and 0. 20 micromol/L, the inhibitive rates were 72. 0% , 65. 1%, 51. 5%, 28.8% ,23.4% respectively; (3) 5-Aza-CdR increased apoptosis rate of tumor cells with a dose and times dependent manner within certain extent, too:at the 1,3,5 d treatment with 5-Aza-CdR 2. 00 micromol/L,the apoptosis rates were 2. 3% ,10. 8 % and 17. 1% ; respectively. (4) 5-Aza-CdR also changed cell cycle of tumor cells: at 24 h treatment with 5-Aza-CdR 2.00 micromol/L,92. 7% tumor cells stopped at S phase and G, phase cells were increased gradually with time. CONCLUSION: DNA promoter hypermethylation is associated with SHP-1 gene silence in Daudi lymphoma cell line. 5-Aza-CdR could effectively cause demethylation and inhibit the growth of tumor cell by reactivating the gene transcription.