Direct-acting Antiviral Drugs Research Articles

Hepatitis C virus-induced differential transcriptional traits in host cells after persistent infection elimination by direct-acting antivirals in cell culture.

Chronic hepatitis C virus infection (HCV) causes liver inflammation and fibrosis, leading to the development of severe liver disease, such as cirrhosis or hepatocellular carcinoma (HCC). Approval of direct-acting antiviral drug combinations has revolutionized chronic HCV therapy, with virus eradication in >98% of the treated patients. The efficacy of these treatments is such that it is formally possible for cured patients to carry formerly infected cells that display irreversible transcriptional alterations directly caused by chronic HCV Infection. Combining differential transcriptomes from two different persistent infection models, we observed a major reversion of infection-related transcripts after complete infection elimination. However, a small number of transcripts were abnormally expressed in formerly infected cells. Comparison of the results obtained in proliferating and growth-arrested cell culture models suggest that permanent transcriptional alterations may be established by several mechanisms. Interestingly, some of these alterations were also observed in the liver biopsies of virologically cured patients. Overall, our data suggest a direct and permanent impact of persistent HCV infection on the host cell transcriptome even after virus elimination, possibly contributing to the development of HCC.

Synthesis and anti-HCV activity of novel 5-arylmethylene-1,3-thiazolidin-4-one derivatives via suppression of NS5B polymerase and COX-2

Hepatitis C (HCV) is a viral infection that leads to forms of acute and chronic liver disease, including cirrhosis (scarring of the liver) and liver cancer. The World Health Organization (WHO) estimated in 2019 that approximately 290,000 people died from hepatitis C (mostly from cirrhosis and hepatocellular carcinoma). Direct-acting antiviral drugs (DAAs) can cure more than 95% of individuals with hepatitis C infection, while research on the discovery of new antiviral agents is still ongoing. The Hepatitis C virus (HCV) can cause various biochemical changes in liver cells, and some of these changes are associated with the COX-2 enzyme. The identification of its role in promoting growth in liver cells as well as its involvement in various cancer types, including hepatocellular carcinoma, has made COX-2 an important target in the development of new agents effective against HCV. In this study, thirty-six new 5-arylmethylene-2-imino-1,3-thiazolidin-4-one derivatives (5a-s, 6a-s) were synthesized through Knoevenagel condensation of 2-[(4-substitutedpyridin-2-yl)imino]-1,3-thiazolidin-4-one derivatives with various aldehydes. Structures of the synthesized compounds were elucidated by the use of spectral and chromatographic techniques, besides elemental analyses. Four compounds were selected for further studies as they were found to suppress the NS5B protein with anti-HCV activity using the Western Blotting method. The selected compounds 5o, 6m, 6r, and 6s inhibited HCV with EC50 values of 8.0 ± 0.2 µM, 13.9 ± 0.45, 9.2 ± 0.2 µM, and 12.1 ± 0.1 µM, respectively. It was determined that these compounds reduced HCV-induced COX-2 promoter activity in Ava5 cells compared to Huh7 cells. The antiviral effects of the compounds were also investigated on DENV, closely related to HCV due to sharing certain biological, structural, and mechanical properties throughout their life cycles. However, no significant effect was observed in the preliminary screening study, indicating the compounds' specificity for HCV. Considering the relationship between HCV, DENV, and COX-2, the compounds' COX-1 and COX-2 enzyme inhibition potentials were investigated both in vitro and in silico. Compounds 6d, 6e, 6f, and 6m, which exhibited high selective COX-2 inhibition, were discussed for their interactions with the active site. Our study revealed that our target compounds suppressed COX-2 both at the protein level and through enzyme inhibition, thus providing promising findings for the discovery of new anti-HCV effective COX-2 inhibitors.

Justification of Intensive care management of children with severe COVID19 (а case study)

the unprecedented COVID-19 pandemic that spread across the world and lasted for more than 3 years had significant medical and social consequences. Despite the fact that in the children's population the coronavirus infection has a milder course and a much lower risk of mortality, there are individual cases with pre-existing comorbidities that cause severe acute respiratory distress syndrome. Thus, in most cases, severe COVID-19 developed in children with pre-existing conditions, including: immunodeficiency, diabetes mellitus, bronchial asthma, etc. Therefore, comorbidities disposed children to progress to severe Covid-19 which requires treatments with more aggressive therapeutic tactics and personalised management. The article presents a clinical case of severe acute respiratory distress syndrome caused by the SARS-CoV-2 virus in an early age child. The patient was observed in the Anesthesiology and Intensive Therapy Department of the Chernivtsi Regional Children's Clinical Hospital. Due to acute respiratory infection, the child has been developing signs of respiratory failure. The available instrumental findings confirmed right lung pneumonia, complicated by pneumothorax. Laboratory tests confirmed leukocytosis, thrombocytopenia, hypertransfusion, increased procalcitonin, increased level of C-reactive protein, D-dimer and interleukin-6 as well as circulating immune complexes. The treatment regimen included respiratory protection (oxygen therapy), support of hemodynamics (infusion of glucose-saline solutions), administration of intravenous normal human immunoglobulin, a short course of parenteral glucocorticosteroids, the use of step-by-step combined antibacterial therapy and a direct-acting antiviral drug (remdesivir), and thromboembolism prophylaxis (low-molecular-weight heparin). Following intensive treatment, the child's condition was characterised by a slow positive dynamics, leading to full recovery. This case demonstrates the need for timely detection of life-threatening conditions caused by COVID-19 and requires an aggressive management of child's condition using a complex intensive therapy.

Comparison of six hepatocellular carcinoma prediction models in Japanese patients after sustained virologic response undergoing rigorous surveillance for hepatocellular carcinoma.

While several predictive models for the development of hepatocellular carcinoma (HCC) have been proposed, including those for patients with chronic hepatitis C virus (HCV) infection who have achieved sustained virologic response (SVR), the best model may differ between regions. We compared the ability of six reported models to stratify the risk of post-SVR HCC in Japan, where rigorous surveillance and early detection of HCC is common. A total of 6048 patients with no history of HCC who achieved SVR by oral direct-acting antiviral drugs were enrolled in this nationwide study. Patients continued HCC surveillance every 6months after SVR. The incidence of post-SVR HCC was compared between risk groups using the aMAP score, FIB-4 index, Tahata model, GAF4 criteria, GES score, and ADRES score. During the observation period with a median duration of 4.0years after SVR, post-SVR HCC developed in 332 patients (5.5%). All six models performed significantly at stratifying the incidence of HCC. However, Harrell's C-index was below 0.8 for all models (range, 0.660-0.748), indicating insufficient stratification ability. Although all six proposed models demonstrated a good ability to predict the development of post-SVR HCC, their ability to stratify the risk of post-SVRHCC was unsatisfactory. Further studies are necessary to identify the best model for assessing the risk of post-SVR HCC in regions where early detection of HCC is common.

Assessment of ubiquitin specific Peptidase-18 gene in peripheral blood of chronic hepatitis C patients treated with direct-acting antiviral drugs

Hepatitis C virus (HCV) infection remains one of the leading causes of liver complications globally. Ubiquitin Specific Peptidase-18 (USP18) is a ubiquitin-specific protease that cleaves interferon-stimulated gene 15 (ISG15) from ISGylated protein complexes and is involved in regulating interferon responsiveness. To study the effect of direct-acting antivirals (DAAs) on the USP18 gene using qPCR, 132 participants were recruited and classified into different groups based on treatment duration. USP18 expression was raised compared to rapid virologic response (RVR) and early virologic response (EVR) groups with P = 0.0026 and P = 0.0016, respectively. USP18 was found to be 7.36 folds higher in naïve patients than those with RVR and sustained viral response (SVR). In RVR and SVR groups where patients had cleared HCV RNA after treatment with direct-acting antiviral agents (DAA) therapy, the expression of USP18 was found to be low, with a fold change of 1.3 and 1.4 folds, respectively. Expression of USP18 was significantly higher in the non-RVR group than in the RVR group. In the No EVR group, gene expression was significantly higher than in the EVR group. It is concluded that targeting HCV proteins using DAAs can cause USP18 expression to be normalized more effectively. Moreover, USP18 is a vital marker indicating treatment resistance and distinguishing responders from non-responders during DAA therapy.

Real-world experience of the Lebanese haemodialysis patient infected with the hepatitis C virus and treated with direct-acting antiviral drugs

Real-world experience of the Lebanese haemodialysis patient infected with the hepatitis C virus and treated with direct-acting antiviral drugs

Insights into unexpected relapse and recovery in HCV-infected patients by studying a stochastic within-host HCV model

The causes of viral rebound leading to disease relapse in some chronic hepatitis C virus-infected patients at the end of treatment with direct-acting antiviral drugs are still unclear, which poses a challenge to further improve the permanent cure rate of HCV-infected patients. In this paper, a stochastic within-host HCV model is studied in detail to gain insights into the phenomena of unexpected relapse and recovery in HCV-infected patients. Theoretically, the existence of an ergodic stationary distribution, which also reflects the long-term coexistence of uninfected hepatocytes and free viruses, is proved by constructing suitable stochastic Lyapunov functions at a threshold condition related to the basic reproduction number of the corresponding deterministic model. In addition, the sufficient conditions for the extinction of free viruses is also given, which implies that the viral load eventually drops to zero and HCV-infected patients are cured. In the numerical simulations, the phenomenon of unexpected relapse and recovery in HCV-infected patients is demonstrated by a few simple examples. By Monte Carlo method, under certain conditions, it is observed that the increase in noise intensities makes the phenomena of unexpected relapse and recovery in HCV-infected patients increase as well.

Recent Advances on the Synthesis of Orally Active Antiviral Drug Molnupiravir

Abstract: Molnupiravir is one of the simple orally active antiviral drugs, sold under the brand name Lagevrio. Initially, this drug was used to treat the influenza virus but later on, used against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This is the first orally active and direct-acting antiviral drug and is highly effective against SARS-CoV- 2. In this review, we discuss different synthetic strategies for the synthesis of molnupiravir, which will help for the further development of novel routes.

The effect of direct-acting antivirals (DAA) on confirmed noninvasive fibrous parameters in chronic hepatitis C patients

Aims: Chronic hepatitis C (CHC) is an important public health problem in terms of the number of people it affects worldwide and the diseases it causes. The high sustained virological response (SVR) rates achieved by the use of direct-acting antiviral (DAA) drugs in the recent period have shown that a new era has begun in this disease. It was aimed to evaluate the effect of DAAs on confirmed noninvasive fibrous parameters together with their effectiveness.
 Methods: 75 patients who were started on DAA treatment for CHC were included in the study. In addition, laboratory parameters values at the beginning of the treatment, 12 and 24 weeks after the end of the treatment, hepatitis C virus ribonucleic acid (HCV RNA) results and Aminotransferase-to-Platelet Ratio Index (APRI), Fibrosis-4 (FIB-4) scores were compared.
 Results: The most common comorbidity in patients is hypertension (HT), and the most common source of transmission is surgical operations. Genotype 1b was the dominant genotype. The SVR rates of all patients 12 and 24 weeks after the end of treatment were 100%. The APRI and FIB-4 scores of the patients decreased significantly at the 12th and 24th weeks at the end of the treatment compared to the beginning of the treatment.
 Conclusion: The confirmed noninvasive fibrous parameters used in the treatment of CHC are useful in evaluating the results of the treatments applied.

The Results of Preclinical Studies Assessing the Effect of Riamilovir on the Reproductive Function and Postnatal Growth of Offspring in Laboratory Animals

The paper presents the results of extended preclinical studies demonstrating the safety using the direct-acting antiviral drug Riamilovir in immature white rats. Riamilovir did not show any toxic effects on the growth of animals, their fertility, behavior, hematological and biochemical parameters, as well as morphology of visceral organs when used in a course. According to the results of the studies, Ramilovir can be recommended for multicenter clinical trials in adolescents and children of younger age groups.

White targeted chromatographic screening method of Molnupiravir and its metabolite with degradation kinetics characterization and in-silico toxicity

SARS-CoV-2 virus triggered a worldwide crisis, with world nations putting up massive efforts to halt its spread. Molnupiravir (MLN) was the first oral, direct-acting antiviral drug approved for nasopharyngeal SARS-CoV-2 infection with favorable safety and tolerability profile. This study aims at determination of MLN and N4-hydroxycytidine (NHC), its main degradation product and its main metabolite, using sensitive, simple, and green HPLC–DAD method. Moreover, under different stress conditions using NaOH, HCl, neutral, H2O2, dry heat and sun light, the method was applied for MLN assay along with kinetics degradation investigation. The linearity range for MLN and NHC were both 0.1–100 µg/mL with LOD and LOQ of 0.013 & 0.043 and 0.003 & 0.011 µg/mL, for MLN and NHC, respectively. MLN was found to be extremely vulnerable to alkali hydrolysis compared with acid and dry heat degradation. In contrast, MLN was stable under conditions of oxidative, neutral, and sunlight-induced deterioration. Acid and alkali-induced degradation followed pseudo first-order kinetics model. In addition, LC–MS-UV was used to suggest the mechanism of the stress-induced degradation route and to characterize the eluted degradation products. Toxicities of both MLN and its degradation products were evaluated using ProTox-II and they were found to be negligibly harmful. The proposed HPLC–DAD was effectively used for the analysis of MLN in commercial pharmaceutical formulations. The proposed method for MLN determination after greenness and whiteness appraisal was found to be superior compared to the reported methods for MLN analysis.

Assessment of hepatic fibrosis, portal hemodynamic changes, and disease severity in patients with HCV-related liver cirrhosis after sustained virologic response to direct-acting antiviral drugs (DAAs)

BackgroundRegression of fibrosis and improvement of portal hemodynamics after achievement of sustained viral response (SVR) in patients with chronic hepatitis C (HCV) is a subject of debate in different studies. Some studies reported improvement in the degree of fibrosis, while others did not find significant changes.ObjectiveWe aimed to evaluate changes in liver fibrosis, portal hemodynamics and clinical outcomes in patients with chronic HCV-related liver cirrhosis after the achievement of SVR with direct-acting antiviral drugs (DAAs).Patients and methodsIn our prospective longitudinal study, a total of 100 patients with chronic HCV infection-related liver cirrhosis were recruited, received DAAs, and completed the follow-up period. Clinical evaluation for assessment of liver disease severity using MELD and Child–Pugh class and scores were done. A noninvasive assessment of liver fibrosis using serum biomarkers (APRI index & FIB4 score) and share wave elastography (SWE) was done. Portal hemodynamic evaluation using Doppler ultrasound was done. All were done at baseline and 3 and 12 months after the end of therapy.ResultsA significant reduction in the degree of fibrosis was observed. Share wave elastography (SWE) readings showed 19.79% and 30.45% reduction 3 and 12 months after the end of therapy respectively (P < 0.001). Regarding the FIB4 score, the percentage of score reduction was 19.8% and 26.46% 3 and 12 months after the end of therapy, respectively (P < 0.01). APRI scores showed 22.6% and 41.09% reduction 3 and 12 months after the end of therapy respectively (P < 0.001). Significant improvement in Child–Pugh scores 3 and 12 months after the end of treatment was observed. Doppler ultrasound showed a significant increase in portal vein flow velocity, a significant decrease in time average mean velocity, and cross-section area 12 months after the end of treatment.ConclusionThere was a considerable degree of reduction of liver fibrosis, improvement of portal hemodynamics, and Child–Pugh score in cirrhotic HCV patients who achieved SVR after DAAs.Trial registrationClinicalTrials.gov, ID: NCT03241823. Registered on 08 May 2017.

Molecular Mechanisms of Resistance to Direct-Acting Antiviral (DAA) Drugs for the Treatment of Hepatitis C Virus Infections.

Hepatitis C virus (HCV) is a hepatotropic virus that affects millions of human lives worldwide. Direct-acting antiviral (DAA) regimens are the most effective HCV treatment option. However, amino acid substitution-dependent resistance to DAAs has been a major challenge. This study aimed to determine the increasing risk of DAA resistance due to substitutions in DAA target non-structural proteins (NS3/4A, NS5A, and NS5B). Using a Sequence Retrieval System (SRS) at the virus pathogen resource (ViPR/BV-BRC), n = 32763 target protein sequences were retrieved and analyzed for resistance-associated amino acid substitutions (RAASs) by the Sequence Feature Variant Type (SFVT) antiviral-resistance assessment modeling tool. Reference target protein sequences with 100% identity were retried from UniProt following NCBI BLAST. The types and locations of RAASs were identified and visualized by AlphaFold and PyMol. Linux-r-base/R-studio was used for the data presentation. Multi-drug-resistant variants of NS3/4A in genotype 1 (n = 9) and genotype 5 (n = 5) along with DAA-specific NS3/4A, NS5A, and NS5B variants were identified pan-genotypically. A total of 27 variants (RAASs) of all the targets were identified. Fourteen genotype 1-specific substitutions: V1196A, V1158I, D1194A/T/G, R1181K, T1080S, Q1106R, V1062A, S1148G, A1182V, Y2065N, M2000T, and L2003V were identified. The most frequent substitutions were V1062L and L2003M, followed by Q2002H. L2003V, Q2002H, M2000T, Y2065N, and NL2003M of NS5A and L2003M of NS5B conferred resistance to daclatasvir. S2702T NS5B was the sofosbuvir-resistant variant. D1194A NS3/4A was triple DAA (simeprevir, faldaprevir, and asunaprevir) resistant. The double-drug resistant variants R1181K (faldaprevir and asunaprevir), A1182V and Q1106K/R (faldaprevir and simeprevir), T1080S (faldaprevir and telaprevir), and single drug-resistant variants V1062L (telaprevir), D1194E/T (simeprevir), D1194G (asunaprevir), S1148A/G (simeprevir), and Q1106L (Boceprevir) of NS3/4A were determined. The molecular phenomenon of DAA resistance is paramount in the development of HCV drug candidates. RAASs in NS3, NS5A, and NS5B reduce the susceptibility to DAAs; therefore, continuous RAAS-dependent resistance profiling in HCV is recommended to minimize the probability of DAA therapeutic failure.

Impact of Treatment of Chronic HCV Patients by Direct Acting Antiviral Drugs (DAADs) on COVID -19 Disease Frequency and Severity

Impact of Treatment of Chronic HCV Patients by Direct Acting Antiviral Drugs (DAADs) on COVID -19 Disease Frequency and Severity

Assessment of Quality of Life of Hepatitis C Virus Patients after Treatment with Direct-Acting Antiviral Drugs

Assessment of Quality of Life of Hepatitis C Virus Patients after Treatment with Direct-Acting Antiviral Drugs

IN HIV-INFECTED IMMUNOLOGICAL NON-RESPONDERS, HEPATITIS C VIRUS ERADICATION CONTRIBUTES TO INCOMPLETE NORMALIZATION OF SYSTEMIC INFLAMMATION INDEXES, BUT DOES NOT LEAD TO RAPID CD4&lt;sup&gt;+&lt;/sup&gt; T-CELL COUNT RECOVERY

In HIV-positive individuals taking antiretroviral therapy, coinfection with hepatitis C virus (HCV) increases the systemic inflammation, which interferes with the CD4+ T-cells regeneration. This study evaluated the effect of HCV eradication on systemic inflammation and CD4+ T-cell regeneration in patients who gave poor response to antiretroviral therapy, the so-called “immunological non-responders” (INRs). HIV-infected patients who received a course of direct-acting antiviral drugs for treating hepatitis C were examined. The control groups included HIV/HCV-coinfected individuals and relatively healthy volunteers. It has been established for the first time that HCV eradication is not accompanied by a complete suppression of systemic inflammation, but improves the T-cell pool composition: in INRs, the blood CD4+/CD8+ T-lymphocyte ratio increases and approaches those of healthy individuals. Apparently, in INRs treated for hepatitis C, the immune system recovery takes time and may be incomplete.

Genotype Distribution and Characteristics of Chronic Hepatitis C Infection in Estonia, Latvia, Lithuania, and Ukraine: The RESPOND-C Study.

Background and objectives: Since 2013, highly effective direct-acting antiviral (DAA) treatment for chronic hepatitis C (CHC) has become available, with cure rates exceeding 95%. For the choice of optimal CHC treatment, an assessment of the hepatitis C virus (HCV) genotype (GT) and liver fibrosis stage is necessary. Information about the distribution of these parameters among CHC patients in Estonia, Latvia, and Lithuania (the Baltic states) and especially in Ukraine is scarce. This study was performed to obtain epidemiologic data regarding CHC GT and fibrosis stage distribution for better planning of resources and prioritization of patients for DAA drug treatment according to disease severity in high-income (the Baltic states) and lower-middle-income (Ukraine) countries. Materials and methods: The retrospective RESPOND-C study included 1451 CHC patients. Demographic and disease information was collected from medical charts for each patient. Results: The most common suspected mode of viral transmission was blood transfusions (17.8%), followed by intravenous substance use (15.7%); however, in 50.9% of patients, the exact mode of transmission was not clarified. In Ukraine (18.4%) and Estonia (26%), transmission by intravenous substance use was higher than in Lithuania (5%) and Latvia (5.3%). Distribution of HCV GT among patients with CHC was as follows: GT1-66.4%; GT3-28.1; and GT2-4.1%. The prevalence of GT1 was the highest in Latvia (84%) and the lowest in Ukraine (63%, p < 0.001). Liver fibrosis stages were distributed as follows: F0-12.2%, F1-26.3%, F2-23.5%, F3-17.1%, and F4-20.9%. Cirrhosis (F4) was more prevalent in Lithuanian patients (30.1%) than in Estonians (8.1%, p < 0.001). Conclusions: This study contributes to the knowledge of epidemiologic characteristics of HCV infection in the Baltic states and Ukraine. The data regarding the patterns of HCV GT and fibrosis stage distribution will be helpful for the development of national strategies to control HCV infection in the era of DAA therapy.

Study of Risk Factors in Patient with Persistant Elevation of Liver Enzymes after Treatment of Chronic HCV by Direct Acting Antiviral Drugs (DAADs)

Abstract Background Hepatitis C virus (HCV) infection is a major global health challenge; it is estimated that more than 80 million people are chronically infected worldwide, with 3- 4 million new infections and 350000 deaths occurring each year because of HCV- related complications. Egypt is the country with the highest HCV prevalence in the world. Aim of the Work The aim of this study is to determine risk factors that cause persistent elevation of liver enzymes in hepatitis c virus patients treated by direct acting antiviral drugs (dyslipidemia, high BMI, type2 diabte, NAFLD). Patients and Methods This retrospective cross sectional study was conducted on 128 Egyptian candidates achieved SVR after 12 weeks with elevated liver enzymes before treatment by (DAAds) Results In this study, we found that there were correlations between the elevated ALT levels and most metabolic risk factors. There were increase in TG, BMI, and decreased. HDL-c, which were components of metabolic syndrome, were associated with high ALT levels in patients. Metabolic syndrome (MS) components are highly prevalent in NAFLD lack of biochemical response was associated with a higher risk to develop liver related complications (decompensation and cancer), indicating that these patients should undergo further hepatological followup. Conclusion Further large-scale studies employing larger number of patients are needed to evaluate risk factors that cause persistent elevated liver enzymes after eradication of hepatitis c viral infection with DAAds.

RESULTS OF ANTIFIBROTIC THERAPY IN PATIENTS WITH CHRONIC HEPATITIS B+C

Modern antiviral therapy regimens for patients with chronic viral hepatitis aim to achieve either long-term suppression of pathogen replication (e.g., nucleoside analogs in chronic hepatitis B) or complete elimination of the virus (such as direct-acting antiviral drugs in chronic hepatitis C). However, antiviral agents do not have a significant impact on the complete restoration of biochemical processes or the prevention of further progression of morphological changes in the liver. These limitations emphasize the ongoing need for new therapeutic strategies that target the processes of fibrogenesis.&#x0D; The aim of the work is to assess the possibility of the effect of the drug "Bicyclol" on fibrotic changes in patients with chronic hepatitis B+C using a non-invasive scale of the rate of fibrosis.&#x0D; Materials and methods&#x0D; An analysis of the dynamics of 62 patients with chronic hepatitis B and C (HCV+C) was conducted. All patients received long-term antiviral therapy consisting of pegylated interferon for 48 weeks. In the main group (Group I), patients with chronic hepatitis B and C were prescribed the drug "Bicyclol" after completing antiviral treatment with interferon. The control group (Group II) followed the principles of proper nutrition and took traditional hepatoprotectors. Based on the identified correlations, a non-invasive scale was proposed to assess the individual risk of liver fibrosis progression.&#x0D; Research results&#x0D; Against the background of the use of an antiviral treatment regimen, the normalization of cytolysis indicators was observed in most patients with CHB+C, but in some patients it was short-lived. Treatment with the drug "Bicyclol" contributed to a further decrease in cytolysis indicators, in the vast majority of patients with CHB+C, the activity of ALT and AST reached the upper limit of the norm. In patients with CHB+C who received only basic treatment without an antifibrotic component, a tendency to maintain an elevated level of transaminases was observed.&#x0D; Conclusions.&#x0D; Thus, the use of the hepatoprotector "Bicyclol" for 12 months ensures a decrease in cytolysis in the liver, is accompanied by a decrease in the severity of fibrosis, and in some patients - its reverse development. The use of the proposed prognostic scale makes it possible to assess the need for early appointment of antifibrotic therapy.

Validation of a Cost-Effective RP-HPLC Method for Quantitative Investigation of Daclatasvir Dihydrochloride in Pharmaceutical Formulations

A well-known direct-acting antiviral (DAA) drug called daclatasvir may be used to treat chronic hepatitis C virus (HCV) infection. Herein, we reported a selective, precise, and a cost-effective analytical method for the measurement of an active pharmaceutical ingredient (API) of daclatasvir dihydrochloride in drug substances as well as drug products via the reversed-phase RP-HPLC technique. To obtain greater separation, the majority of the chromatographic conditions were improved. Best separation findings were achieved under chromatographic conditions with an HPLC column of USP L1 (150 × 4.6 mm, 5 μm) by utilizing a combination of acetonitrile and buffer solution of KH2PO4 (30 : 70, v/v) as a mobile phase at a stream rate of 1 mL.min−1 with a finding at 300 nm and a column temperature of 40°C. Linearity was examined in the range of 90–210 ppm (R2 = 0.999) for daclatasvir dihydrochloride. The new technique has been verified using industry-recognized criteria, including applicability, system precision, accuracy, robustness, specificity, range, linearity, quantification limit, reagent stability, and detection limit. All the measured metrics were determined to be within acceptable limits using the criteria of the Worldwide Council for Harmonisation (ICH). In pharmaceutical labs, daclatasvir dihydrochloride may be analyzed qualitatively and quantitatively using the well-established RP-HPLC technique. Our study also highlights the need to evaluate the greenness of the method developed using a recognized tool ,i.e., Analytical Greenness Metrics (AGREE).