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PURPOSE: To report 2-year results from the Archway clinical trial of the Port Delivery System with ranibizumab (PDS) for treatment of neovascular age-related macular degeneration (nAMD). DESIGN: Phase 3, randomized, multicenter, open-label, active-comparator-controlled trial. PARTICIPANTS: Patients with previously treated nAMD diagnosed within 9 months of screening and responsive to anti-vascular endothelial growth factor therapy. METHODS: Patients were randomized 3:2 to PDS with ranibizumab 100 mg/ml with fixed refill-exchanges every 24 weeks (PDS Q24W) or intravitreal ranibizumab 0.5 mg injections every 4 weeks (monthly ranibizumab). Patients were followed through 4 complete refill-exchange intervals (â¼2 years). MAIN OUTCOME MEASURES: Change in best-corrected visual acuity (BCVA) Early Treatment Diabetic Retinopathy Study (ETDRS) letter score from baseline averaged over weeks 44 and 48, weeks 60 and 64, and weeks 88 and 92 (noninferiority margin, -3.9 ETDRS letters). RESULTS: The PDS Q24W was noninferior to monthly ranibizumab, with differences in adjusted mean change in BCVA score from baseline averaged over weeks 44/48, 60/64 and 88/92 of -0.2 (95% confidence interval [CI], -1.8 to +1.3), +0.4 (95% CI, -1.4 to +2.1) and -0.6 ETDRS letters (95% CI, -2.5 to +1.3), respectively. Anatomic outcomes were generally comparable between arms through week 96. Through each of 4 PDS refill-exchange intervals, 98.4%, 94.6%, 94.8%, and 94.7% of PDS Q24W patients assessed did not receive supplemental ranibizumab treatment. The PDS ocular safety profile was generally unchanged from primary analysis. Prespecified ocular adverse events of special interest (AESI) were reported in 59 (23.8%) PDS and 17 (10.2%) monthly ranibizumab patients. The most common AESI reported in both arms was cataract (PDS Q24W, 22 [8.9%]; monthly ranibizumab, 10 [6.0%]). Events in the PDS Q24W arm included (patient incidence) 10 (4.0%) conjunctival erosions, 6 (2.4%) conjunctival retractions, 4 (1.6%) endophthalmitis cases, and 4 (1.6%) implant dislocations. Serum ranibizumab sampling showed that the PDS continuously released ranibizumab over the 24-week refill-exchange interval and ranibizumab serum concentrations were within the range experienced with monthly ranibizumab. CONCLUSIONS: The PDS Q24W showed noninferior efficacy to monthly ranibizumab through approximately 2 years, with approximately 95% of PDS Q24W patients not receiving supplemental ranibizumab treatment in each refill-exchange interval. The AESIs were generally manageable, with learnings continually implemented to minimize PDS-related AEs. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.
Sujet(s)
Rétinopathie diabétique , Dégénérescence maculaire , Dégénérescence maculaire humide , Humains , Ranibizumab/usage thérapeutique , Inhibiteurs de l'angiogenèse , Acuité visuelle , Rétinopathie diabétique/traitement médicamenteux , Dégénérescence maculaire/traitement médicamenteux , Injections intravitréennes , Résultat thérapeutique , Dégénérescence maculaire humide/diagnostic , Dégénérescence maculaire humide/traitement médicamenteux , Dégénérescence maculaire humide/induit chimiquementRÉSUMÉ
INTRODUCTION: Immunocompromised patients infected with influenza exhibit prolonged viral shedding and higher risk of resistance. Optimized treatment strategies are needed to reduce the risk of antiviral resistance. This phase IIIb, randomized, double-blind study (NCT00545532) evaluated conventional-dose or double-dose oseltamivir for the treatment of influenza in immunocompromised patients. METHODS: Patients with primary or secondary immunodeficiency and influenza infection were randomized 1:1 to receive conventional-dose oseltamivir (75 mg adolescents/adults [≥ 13 years]; 30-75 mg by body weight in children [1-12 years]) or double-dose oseltamivir (150 or 60-150 mg, respectively), twice daily for an extended period of 10 days. Nasal/throat swabs were taken for virology assessments at all study visits. Co-primary endpoints were safety/tolerability and viral resistance. Secondary endpoints included time to symptom alleviation (TTSA) and time to cessation of viral shedding (TTCVS). RESULTS: Of 228 patients enrolled between February 2008 and May 2017, 215 (199 adults) were evaluable for safety, 167 (151 adults) for efficacy, and 152 (138 adults) for resistance. Fewer patients experienced an adverse event (AE) in the conventional-dose group (50.5%) versus the double-dose group (59.1%). The most frequently reported AEs were nausea, diarrhea, vomiting, and headache. Fifteen patients had post-baseline resistance, more commonly in the conventional-dose group (n = 12) than in the double-dose group (n = 3). In adults, median TTSA was similar between arms, while median TTCVS was longer with conventional dosing. CONCLUSIONS: Oseltamivir was well tolerated, with a trend toward better safety/tolerability for conventional dosing versus double dosing. Resistance rates were higher with conventional dosing in this immunocompromised patient population. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT00545532. FUNDING: F. Hoffmann-La Roche Ltd.
RÉSUMÉ
Current standard-of-care treatment of chronically infected hepatitis C virus (HCV) patients involves direct-acting antivirals (DAA). However, concerns exist regarding the emergence of drug -resistant variants and subsequent treatment failure. In this study, we investigate potential natural drug-resistance mutations in the NS5B gene of HCV genotype 1b from treatment-naïve patients. Population-based sequencing and 454 deep sequencing of NS5B gene were performed on plasma and liver samples obtained from 18 treatment- naïve patients. The quasispecies distribution in plasma and liver samples showed a remarkable overlap in each patient. Although unique sequences in plasma or liver were observed, in the majority of cases the most dominant sequences were shown to be identical in both compartments. Neither in plasma nor in the liver codon changes were detected at position 282 that cause resistance to nucleos(t)ide analogues. However, in 10 patients the V321I change conferring resistance to nucleos(t)ide NS5B polymerase inhibitors and in 16 patients the C316N/Y/H non-nucleoside inhibitors were found mainly in liver samples. In conclusion, 454-deep sequencing of liver and plasma compartments in treatment naïve patients provides insight into viral quasispecies and the pre-existence of some drug-resistant variants in the liver, which are not necessarily present in plasma.
Sujet(s)
Résistance virale aux médicaments , Hépatite C chronique/génétique , Hépatite C/virologie , Quasi-espèce , Protéines virales non structurales/génétique , Antiviraux/pharmacologie , Génotype , Séquençage nucléotidique à haut débit/méthodes , Humains , Foie/virologie , Plasma sanguin/virologie , Analyse de séquence d'ARNRÉSUMÉ
BACKGROUND AND AIM: Chronic hepatitis C is an important public health problem in Asia. We evaluated the safety, efficacy, and pharmacokinetics of fixed-dose ritonavir-boosted danoprevir plus peginterferon alfa-2a/ribavirin in treatment-naive Asian patients with chronic hepatitis C virus (HCV) genotype (G)1 infection. METHODS: Treatment-naive G1 patients in Taiwan, Thailand, and Korea with serum HCV-RNA level ≥ 105 IU/mL received ritonavir-boosted danoprevir 125/100 mg twice daily plus peginterferon alfa-2a/ribavirin for either 12 (noncirrhotic patients: Arm A, n = 34) or 24 weeks (cirrhotic patients: Arm B, n = 27) in this phase II open-label study. Sustained virologic response was defined as HCV-RNA < 25 IU/mL 12 weeks after end of treatment (SVR12). RESULTS: Similar SVR12 rates were achieved in Arms A (88.2%; 95% confidence interval, 73.4-95.3%) and B (88.9%; 71.9-96.2%). Most patients had G1b infection, among whom SVR12 rates in Arms A and B were 96.7% and 91.7%, respectively. The overall SVR12 rate was 94.0% in noncirrhotic Taiwanese patients (100% in the subset of G1b patients). No patients withdrew for safety reasons. Three (11%) cirrhotic patients (Arm B) experienced serious adverse events, none of which was considered to be related to treatment. No Grade 3/4 alanine aminotransferase elevations were reported. The pharmacokinetic properties of danoprevir were broadly overlapping in noncirrhotic and cirrhotic patients both on Days 1 and 14. CONCLUSIONS: Ritonavir-boosted danoprevir plus peginterferon alfa-2a/ribavirin produced sustained virologic response rates > 90% after 12 weeks' treatment in noncirrhotic and 24 weeks' treatment in cirrhotic Asian patients with G1b infection and was well tolerated. These regimens are well suited to countries where G1b predominates.
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Antiviraux/usage thérapeutique , Hépatite C chronique/traitement médicamenteux , Cirrhose du foie/traitement médicamenteux , Adulte , Sujet âgé , Antiviraux/effets indésirables , Antiviraux/sang , Cyclopropanes , Association de médicaments , Femelle , Hepacivirus/génétique , Hepacivirus/isolement et purification , Hépatite C chronique/complications , Humains , Interféron alpha/effets indésirables , Interféron alpha/sang , Interféron alpha/usage thérapeutique , Isoindoles , Lactames/effets indésirables , Lactames/sang , Lactames/usage thérapeutique , Lactames macrocycliques , Cirrhose du foie/virologie , Mâle , Adulte d'âge moyen , Polyéthylène glycols/effets indésirables , Polyéthylène glycols/usage thérapeutique , Proline/analogues et dérivés , ARN viral/sang , Protéines recombinantes/effets indésirables , Protéines recombinantes/sang , Protéines recombinantes/usage thérapeutique , Ribavirine/effets indésirables , Ribavirine/sang , Ribavirine/usage thérapeutique , Ritonavir/effets indésirables , Ritonavir/sang , Ritonavir/usage thérapeutique , Sulfonamides/effets indésirables , Sulfonamides/sang , Sulfonamides/usage thérapeutique , Jeune adulteRÉSUMÉ
BACKGROUND AND AIM: Effective and safe antiviral treatment regimens are needed for patients with chronic hepatitis C (CHC) and cirrhosis. METHODS: An international open-label trial was conducted in CHC patients with genotype (G)1/4 infection, compensated cirrhosis, HCV RNA ≥ 50,000 IU/mL and body mass index 18-35 kg/m(2). Treatment-naive patients (Cohort 1) received a triple therapy regimen [danoprevir/r 100/100 mg twice daily (bid), ribavirin 1000/1200 mg/day and peginterferon alfa-2a 180 µg/week] for 24 weeks. Prior null responders (Cohort 2) received a quadruple therapy regimen (danoprevir/r 100/100 mg bid, mericitabine 1000 mg bid and peginterferon alfa-2a/ribavirin). The primary efficacy outcome was sustained virological response (HCV RNA < limit of quantification, target not detected) at end of the 24-week follow-up period (SVR24). RESULTS: In Cohort 1 (n = 23), 73.9 and 65.2 % of patients had a virological response at Weeks 4 and 24, respectively; 39.1 % achieved SVR24 (G1a = 1/13; G1b = 8/9; G4 = 0/1). In Cohort 2 (n = 20), 100 % achieved virological response at Weeks 4 and 24; 65 % achieved SVR24 (G1a = 4/8; G1b = 7/10; G4 = 2/2). Treatment failure was more common in G1a than G1b-infected patients and less common in patients receiving quadruple therapy. Treatment failure was associated with emergence of resistance to danoprevir, but not mericitabine. The safety profile was typical of that associated with peginterferon alfa-2a/ribavirin. No deaths/episodes of hepatic decompensation occurred. CONCLUSIONS: Treatment with danoprevir/r-based regimens for 24 weeks is safe and well tolerated in CHC patients with compensated cirrhosis. A quadruple therapy regimen (danoprevir/r, mericitabine, peginterferon alfa/ribavirin) produced high SVR24 rates in prior null responders, particularly among G1b patients.
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Antiviraux/usage thérapeutique , Hepacivirus/effets des médicaments et des substances chimiques , Hépatite C/traitement médicamenteux , Lactames/usage thérapeutique , Cirrhose du foie/étiologie , Ritonavir/usage thérapeutique , Sulfonamides/usage thérapeutique , Antiviraux/administration et posologie , Cyclopropanes , Calendrier d'administration des médicaments , Femelle , Hepacivirus/génétique , Hépatite C/complications , Hépatite C/virologie , Humains , Interféron alpha/administration et posologie , Interféron alpha/usage thérapeutique , Isoindoles , Lactames/administration et posologie , Lactames macrocycliques , Cirrhose du foie/virologie , Mâle , Adulte d'âge moyen , Polyéthylène glycols/administration et posologie , Polyéthylène glycols/usage thérapeutique , Proline/analogues et dérivés , Protéines recombinantes/administration et posologie , Protéines recombinantes/usage thérapeutique , Ribavirine/administration et posologie , Ribavirine/usage thérapeutique , Ritonavir/administration et posologie , Sulfonamides/administration et posologie , Échec thérapeutiqueRÉSUMÉ
UNLABELLED: Most patients with chronic hepatitis C virus (HCV) genotype 1 infection who have had a previous null response (<2-log10 reduction in HCV RNA by treatment week 12) to peginterferon/ribavirin (PegIFN/RBV) do not achieve a sustained virological response (SVR) when re-treated with a first-generation HCV protease inhibitor (PI) administered in combination with PegIFN/RBV. We studied the incremental benefits associated with adding mericitabine (nucleoside analog inhibitor of HCV polymerase) to PI plus PegIFN alfa-2a/RBV-based therapy in two double-blind randomized multicenter phase 2 trials (with boceprevir in DYNAMO 1, and with telaprevir in DYNAMO 2). The primary endpoint in both trials was SVR, defined as HCV RNA <25 IU/mL 12 weeks after the end of treatment (SVR12). Overall, the addition of mericitabine to PI plus PegIFN alfa-2a/RBV therapy resulted in SVR12 rates of 60-70% in DYNAMO 1 and of 71-96% in DYNAMO 2. SVR12 rates were similar in patients infected with HCV genotype 1a and 1b in both trials. The placebo control arms in both studies were stopped because of high rates of virological failure. Numerically lower relapse rates were associated with longer treatment with mericitabine (24 versus 12 weeks), telaprevir-containing regimens, and regimens that included 48 weeks of PegIFN alfa-2a/RBV therapy. No mericitabine resistance mutations were identified in any patient in either trial. The addition of mericitabine did not add to the safety burden associated with either telaprevir or boceprevir-based regimens. These studies demonstrate increased SVR rates and reduced relapse rates in difficult-to-treat patients when a nucleoside polymerase inhibitor with intermediate antiviral potency is added to regimens containing a first-generation PI. TRIAL REGISTRATION: ClinicalTrials.gov NCT01482403 and ClinicalTrials.gov NCT01482390.
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Antiviraux/usage thérapeutique , Désoxycytidine/analogues et dérivés , Hépatite C chronique/traitement médicamenteux , Interféron alpha/usage thérapeutique , Oligopeptides/usage thérapeutique , Polyéthylène glycols/usage thérapeutique , Proline/analogues et dérivés , Ribavirine/usage thérapeutique , Adulte , Anticorps antiviraux/biosynthèse , Anticorps antiviraux/sang , Antimétabolites/usage thérapeutique , Désoxycytidine/usage thérapeutique , Méthode en double aveugle , Calendrier d'administration des médicaments , Association de médicaments/méthodes , Femelle , Génotype , Hepacivirus/effets des médicaments et des substances chimiques , Hepacivirus/enzymologie , Hepacivirus/croissance et développement , Hépatite C chronique/immunologie , Hépatite C chronique/anatomopathologie , Hépatite C chronique/virologie , Humains , Mâle , Adulte d'âge moyen , Proline/usage thérapeutique , Inhibiteurs de protéases/usage thérapeutique , ARN viral/antagonistes et inhibiteurs , ARN viral/génétique , Protéines recombinantes/usage thérapeutique , Résultat thérapeutiqueRÉSUMÉ
BACKGROUND & AIMS: Setrobuvir is a direct-acting antiviral (DAA) non-nucleoside inhibitor of hepatitis C virus (HCV) polymerase. This study examined interferon-free combinations containing setrobuvir, a ritonavir-boosted protease inhibitor (danoprevir/r) and ribavirin, with/without the nucleoside inhibitor mericitabine in HCV genotype (G)1 patients. METHODS: Non-cirrhotic treatment-naïve patients (N = 110) were randomized to five groups. Three groups received a 14-day mericitabine/ribavirin lead-in followed by treatment with 3 DAAs (setrobuvir, danoprevir/r, mericitabine) plus ribavirin for 12 weeks (Group A: G1a; D: G1b) or 24 weeks (B: G1a), and two groups received 2 DAAs (setrobuvir, danoprevir/r) plus ribavirin for 12 weeks (E: G1b) or 24 weeks (C: G1a). Efficacy was defined as sustained virological response (HCV RNA <25 IU/ml after 12 weeks' follow-up, SVR12). RESULTS: Two groups met predefined futility criteria for breakthrough (C) or relapse (A) and were discontinued. SVR12 rates were 42.9% (3/7) and 74.1% (20/27) in G1a patients in Groups A and B, respectively, and 95.7% (22/23) and 68.2% (15/22) in G1b patients in Groups D and E respectively. All G1a patients assigned to 24 weeks of treatment who experienced a decrease in HCV RNA of ≥2.3 log10 IU by the end of the lead-in period (n = 28) achieved SVR12. Overall, treatment was well tolerated and most adverse events were mild to moderate. No major safety signals were identified. CONCLUSIONS: An interferon-free setrobuvir-based regimen (3 DAAs plus ribavirin) is safe and effective in treatment-naïve G1 patients.
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Antiviraux/usage thérapeutique , Benzothiadiazines/usage thérapeutique , Hepacivirus/effets des médicaments et des substances chimiques , Hépatite C chronique/traitement médicamenteux , Interférons/usage thérapeutique , Quinolinone/usage thérapeutique , Adulte , Antiviraux/effets indésirables , Australie , Benzothiadiazines/effets indésirables , Cyclopropanes , Désoxycytidine/analogues et dérivés , Désoxycytidine/usage thérapeutique , Association de médicaments , Europe , Femelle , Génotype , Hepacivirus/génétique , Hépatite C chronique/diagnostic , Humains , Interférons/effets indésirables , Isoindoles , Lactames/usage thérapeutique , Lactames macrocycliques , Mâle , Adulte d'âge moyen , Nouvelle-Zélande , Phénotype , Proline/analogues et dérivés , Quinolinone/effets indésirables , ARN viral/sang , Induction de rémission , Ribavirine/usage thérapeutique , Sulfonamides/usage thérapeutique , Facteurs temps , Résultat thérapeutique , États-Unis , Charge viraleRÉSUMÉ
BACKGROUND & AIMS: Danoprevir is a hepatitis C virus (HCV) protease inhibitor with activity against genotypes (G)1/G4, which is maintained at lower doses by ritonavir-boosting. We report results of a large, randomized, active-controlled phase IIb study of ritonavir-boosted danoprevir (danoprevir/r) plus peginterferon alpha-2a/ribavirin (P/R) in treatment-naive patients with HCV G1/4 infection. METHODS: Treatment-naive patients with HCV G1/4 infection were randomized to twice-daily danoprevir/r 200/100 mg (A, n = 92); 100/100 mg (B, n = 93); or 50/100 mg (C, n = 94) plus P/R for 24 weeks; twice-daily danoprevir/r 100/100 mg (D, n = 94) plus P/R for 12 or 24 weeks; or P/R alone (E, n = 44) for 48 weeks. Patients in the response-guided therapy arm (D) with an extended rapid virological response (eRVR2: HCV RNA <15 IU/ml during Weeks 2-10) stopped all therapy at Week 12; non-eRVR2 patients continued all treatment to Week 24. The primary efficacy endpoint was sustained the virological response (SVR24: HCV RNA <15 IU/ml after 24 weeks of untreated follow-up). RESULTS: SVR24 rates in Arms A, B, C, D and E were 89.1%, 78.5%, 66.0%, 69.1% and 36.4%, respectively, in the overall population; 83.6%, 69.6%, 60.3%, 59.2% and 38.5% in G1a-infected patients, 96.6%, 93.1%, 73.1%, 78.4% and 28.6% in G1b-infected patients and 100%, 87.5%, 100%, 100% and 66.7% in G4-infected patients. Danoprevir/r plus P/R was generally well tolerated compared with P/R alone. There was a higher incidence of serious adverse events in danoprevir-treatment arms, but most were associated with P/R. CONCLUSIONS: The combination of danoprevir/r plus P/R is efficacious in treatment-naïve patients with HCV genotype 1 or 4 infection.
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Hepacivirus/génétique , Hépatite C/traitement médicamenteux , Interféron alpha/usage thérapeutique , Lactames/usage thérapeutique , Polyéthylène glycols/usage thérapeutique , Ritonavir/usage thérapeutique , Sulfonamides/usage thérapeutique , Adulte , Cyclopropanes , Association de médicaments/méthodes , Femelle , Génotype , Hepacivirus/effets des médicaments et des substances chimiques , Hépatite C/génétique , Humains , Isoindoles , Lactames macrocycliques , Mâle , Proline/analogues et dérivés , ARN viral/sang , Protéines recombinantes/usage thérapeutiqueRÉSUMÉ
BACKGROUND & AIMS: Chronic hepatitis C treatment for prior non-responders to peginterferon (PegIFN)/ribavirin remains suboptimal. The MATTERHORN study evaluated regimens containing ritonavir-boosted danoprevir (danoprevir/r) in prior PegIFN alfa/ribavirin non-responders. METHODS: Prior partial responders (N=152) were randomized to 24 weeks of twice-daily danoprevir/r 100/100mg, mericitabine 1000 mg and ribavirin 1000/1200 mg (IFN-free); danoprevir/r plus PegIFN alfa-2a/ribavirin (triple); or danoprevir/r, mericitabine and PegIFN alfa-2a/ribavirin (Quad). Prior null responders (N=229) were randomized to 24 weeks of IFN-free therapy, or quad alone (Quad 24) or quad plus 24-weeks of PegIFN alfa-2a/ribavirin (Quad 48). The primary endpoint was sustained virological response (HCV RNA <25 IU/ml) 24 weeks after end-of-treatment (SVR24). Due to high relapse rates, genotype (G) 1a patients in IFN-free arms were offered additional PegIFN alfa-2a/ribavirin. RESULTS: Among prior partial responders, SVR24 rates were 46.2%, 51.0%, and 86.0%, in the IFN-free, Triple and Quad arms, respectively; among prior null responders, SVR24 rates were 45.5%, 80.5%, and 83.8% respectively. Relapse rates were lower and SVR24 rates higher in G1b-infected than G1a-infected patients. SVR24 rates in G1a and G1b patients randomized to Quad were 75.0% and 96.2%, respectively, in the partial Quad arm, and 68.1% and 100%, respectively, in the null Quad 24 arm. Treatment failure was associated with resistance to danoprevir, but not to mericitabine, and was more common in G1a infected patients. Treatment was well-tolerated. CONCLUSIONS: Danoprevir/r, mericitabine plus PegIFN alfa-2a/ribavirin was well-tolerated and produced high overall SVR24 rates in prior partial and null responders to PegIFN alfa/ribavirin. In contrast, IFN-free regimens were associated with unacceptably high relapse rates.
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Hepacivirus/génétique , Hépatite C chronique/traitement médicamenteux , Interféron alpha/administration et posologie , Lactames/administration et posologie , Polyéthylène glycols/administration et posologie , ARN viral/génétique , Ribavirine/administration et posologie , Ritonavir/administration et posologie , Sulfonamides/administration et posologie , Cyclopropanes , Inhibiteurs du cytochrome P-450 CYP3A/administration et posologie , Relation dose-effet des médicaments , Calendrier d'administration des médicaments , Vecteurs de médicaments , Association de médicaments , Femelle , Études de suivi , Génotype , Hépatite C chronique/virologie , Humains , Isoindoles , Lactames macrocycliques , Mâle , Adulte d'âge moyen , Proline/analogues et dérivés , Protéines recombinantes/administration et posologie , Études rétrospectives , Résultat thérapeutiqueRÉSUMÉ
BACKGROUND AND OBJECTIVES: Hepatitis C virus (HCV) variants that confer resistance to direct-acting-antiviral agents (DAA) have been detected by standard sequencing technology in genotype (G) 1 viruses from DAA-naive patients. It has recently been shown that virological response rates are higher and breakthrough rates are lower in G1b infected patients than in G1a infected patients treated with certain classes of HCV DAAs. It is not known whether this corresponds to a difference in the composition of G1a and G1b HCV quasispecies in regards to the proportion of naturally occurring DAA-resistant variants before treatment. METHODS: We used ultradeep pyrosequencing to determine the prevalence of low-abundance (<25% of the sequence reads) DAA-resistant variants in 191 NS3 and 116 NS5B isolates from 208 DAA-naive G1-infected patients. RESULTS: A total of 3.5 million high-quality reads of ≥ 200 nucleotides were generated. The median coverage depth was 4150x and 4470x per NS3 and NS5B amplicon, respectively. Both G1a and G1b populations showed Shannon entropy distributions, with no difference between G1a and G1b in NS3 or NS5B region at the nucleotide level. A higher number of substitutions that confer resistance to protease inhibitors were observed in G1a isolates (mainly at amino acid 80 of the NS3 region). The prevalence of amino acid substitutions that confer resistance to NS5B non-nucleoside inhibitors was similar in G1a and G1b isolates. The NS5B S282T variant, which confers resistance to the polymerase inhibitors mericitabine and sofosbuvir, was not detected in any sample. CONCLUSION: The quasispecies genetic diversity and prevalence of DAA-resistant variants was similar in G1a and G1b isolates and in both NS3 and NS5B regions, suggesting that this is not a determinant for the higher level of DAA resistance observed across G1a HCV infected patients upon treatment.
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Résistance virale aux médicaments/génétique , Fréquence d'allèle , Génotype , Hepacivirus/génétique , Hépatite C/virologie , Séquence nucléotidique , Hepacivirus/effets des médicaments et des substances chimiques , Hepacivirus/isolement et purification , Séquençage nucléotidique à haut débit , Humains , Données de séquences moléculaires , Analyse de séquence d'ADN , Protéines virales non structurales/génétiqueRÉSUMÉ
Mapping protein-protein interactions is essential to fully characterize the biological function of a protein and improve our understanding of diseases. Affinity purification coupled to mass spectrometry (AP-MS) using selective antibodies against a target protein has been commonly applied to study protein complexes. However, one major limitation is a lack of specificity as a substantial part of the proposed binders is due to nonspecific interactions. Here, we describe an innovative immuno-competitive capture mass spectrometry (ICC-MS) method to allow systematic investigation of protein-protein interactions. ICC-MS markedly increases the specificity of classical immunoprecipitation (IP) by introducing a competition step between free and capturing antibody prior to IP. Instead of comparing only one experimental sample with a control, the methodology generates a 12-concentration antibody competition profile. Label-free quantitation followed by a robust statistical analysis of the data is then used to extract the cellular interactome of a protein of interest and to filter out background proteins. We applied this new approach to specifically map the interactome of hepatitis C virus (HCV) nonstructural protein 5A (NS5A) in a cellular HCV replication system and uncovered eight new NS5A-interacting protein candidates along with two previously validated binding partners. Follow-up biological validation experiments revealed that large tumor suppressor homolog 1 and 2 (LATS1 and LATS2, respectively), two closely related human protein kinases, are novel host kinases responsible for NS5A phosphorylation at a highly conserved position required for optimal HCV genome replication. These results are the first illustration of the value of ICC-MS for the analysis of endogenous protein complexes to identify biologically relevant protein-protein interactions with high specificity.
Sujet(s)
Hepacivirus/croissance et développement , Cartographie d'interactions entre protéines/méthodes , Protein-Serine-Threonine Kinases/métabolisme , Protéines suppresseurs de tumeurs/métabolisme , Protéines virales non structurales/métabolisme , Lignée cellulaire , Réplication de l'ADN/génétique , Génome viral/génétique , Humains , Spectrométrie de masse/méthodes , Antigènes mineurs d'histocompatibilité , Phosphorylation , Phosphotransferases (Alcohol Group Acceptor)/génétique , Liaison aux protéines , Protein-Serine-Threonine Kinases/génétique , Structure tertiaire des protéines , Protéome/analyse , Interférence par ARN , Petit ARN interférent , Protéines suppresseurs de tumeurs/génétique , Réplication virale/génétique , Réplication virale/physiologieRÉSUMÉ
Danoprevir (DNV) is a hepatitis C virus (HCV) protease inhibitor that achieves high sustained virologic response (SVR) rates in combination with peginterferon alfa-2a-ribavirin in treatment-naive HCV genotype 1 (G1)-infected patients. This study explored the efficacy and safety of ritonavir-boosted DNV (DNVr) plus peginterferon alfa-2a-ribavirin in G1-infected prior peginterferon-ribavirin null responders. Null responders (<2-log10 reduction in HCV RNA level at week 12) were given an open-label combination of 100 mg of ritonavir and 100 mg of DNV (100/100 mg DNVr) every 12 h (q12h) plus peginterferon alfa-2a-ribavirin for 12 weeks. All patients achieving an early virologic response (EVR; ≥2-log10 decrease in HCV RNA by week 12) continued treatment with peginterferon alfa-2a-ribavirin; those without an EVR discontinued all study drugs. Twenty-four prior null responders were enrolled; 16 patients (67%) were infected with HCV G1b, and 8 (33%) were infected with G1a. Ninety-six percent of patients had an IL28B non-CC genotype. A sustained virologic response at 24 weeks posttreatment (SVR24) was achieved in 67% of patients, with a higher rate in G1b-infected (88%) than G1a-infected (25%) patients. Resistance-related breakthrough occurred in 4/8 G1a and 1/16 G1b patients through the DNV resistance-associated variant (RAV) NS3 R155K. NS3 R155K was also detected in 2/2 G1a patients who relapsed. Treatment was well tolerated. Two patients withdrew prematurely from study medications due to adverse events. Two serious adverse events were reported; both occurred after completion of DNVr therapy and were considered unrelated to treatment. No grade 3 or 4 alanine aminotransferase (ALT) elevations were observed. DNVr plus peginterferon alfa-2a-ribavirin demonstrated high SVR24 rates in HCV G1b-infected prior null responders and was well tolerated. (This study has been registered at ClinicalTrials.gov under registration no. NCT01185860.).
Sujet(s)
Antiviraux/administration et posologie , Hépatite C chronique/traitement médicamenteux , Interféron alpha/administration et posologie , Lactames/administration et posologie , Polyéthylène glycols/administration et posologie , ARN viral/antagonistes et inhibiteurs , Ribavirine/administration et posologie , Ritonavir/administration et posologie , Sulfonamides/administration et posologie , Adulte , Alanine transaminase/sang , Antiviraux/effets indésirables , Cyclopropanes , Calendrier d'administration des médicaments , Résistance virale aux médicaments/effets des médicaments et des substances chimiques , Association de médicaments , Femelle , Génotype , Hepacivirus/effets des médicaments et des substances chimiques , Hepacivirus/physiologie , Hépatite C chronique/virologie , Humains , Interféron alpha/effets indésirables , Isoindoles , Lactames/effets indésirables , Lactames macrocycliques , Mâle , Adulte d'âge moyen , Mutation , Polyéthylène glycols/effets indésirables , Proline/analogues et dérivés , ARN viral/génétique , Protéines recombinantes/administration et posologie , Protéines recombinantes/effets indésirables , Ribavirine/effets indésirables , Ritonavir/effets indésirables , Sulfonamides/effets indésirables , Résultat thérapeutique , Charge virale/effets des médicaments et des substances chimiques , Protéines virales non structurales/génétiqueRÉSUMÉ
BACKGROUND: Resistance to mericitabine (prodrug of HCV NS5B polymerase inhibitor PSI-6130) is rare and conferred by the NS5B S282T mutation. METHODS: Serum HCV RNA from patients who experienced viral breakthrough, partial response, or nonresponse in 2 clinical trials in which patients received mericitabine plus peginterferon alfa-2a (40KD)/ribavirin were analyzed by population and clonal sequence analysis as well as phenotypic assay for assessment of in vivo mericitabine resistance. RESULTS: Among 405 patients treated with mericitabine plus peginterferon alfa-2a/ribavirin in PROPEL and JUMP-C, virologic breakthrough or nonresponse were not observed; 12 patients experienced a partial response. The NS5B S282T resistance mutation was not observed in any patient. A number of treatment-associated NS5B changes were observed and characterized. A novel double mutant (L159F/L320F) with impaired replication capacity was detected in one HCV genotype 1b-infected patient. Introduction of double mutant L159F/L320F into genotype 1a (H77) and 1b (Con-1) replicons, respectively, increased the EC50 for mericitabine by 3.1- and 5.5-fold and the EC90 by 3.1- and 8.9-fold. The double mutant also decreased susceptibility to sofosbuvir (GS-7977) and GS-938 but not setrobuvir, relative to wild-type. CONCLUSIONS: A novel and replication-deficient double mutation (L159F/L320F) confers low-level resistance to mericitabine and cross-resistance to both sofosbuvir and GS-938. CLINICAL TRIALS REGISTRATION: NCT00869661, NCT01057667.
Sujet(s)
Antiviraux/usage thérapeutique , Désoxycytidine/analogues et dérivés , Hépatite C chronique/traitement médicamenteux , Mutation/effets des médicaments et des substances chimiques , Uridine monophosphate/analogues et dérivés , Protéines virales non structurales/antagonistes et inhibiteurs , Désoxycytidine/usage thérapeutique , Résistance virale aux médicaments/effets des médicaments et des substances chimiques , Résistance virale aux médicaments/génétique , Association de médicaments/méthodes , Génotype , Hepacivirus/effets des médicaments et des substances chimiques , Hepacivirus/génétique , Hépatite C chronique/sang , Hépatite C chronique/génétique , Humains , Interféron alpha/usage thérapeutique , Mutation/génétique , Polyéthylène glycols/usage thérapeutique , ARN viral/sang , ARN viral/effets des médicaments et des substances chimiques , ARN viral/génétique , Protéines recombinantes/usage thérapeutique , Réplicon/effets des médicaments et des substances chimiques , Réplicon/génétique , Ribavirine/usage thérapeutique , Sofosbuvir , Uridine monophosphate/usage thérapeutiqueRÉSUMÉ
In the past few years, there have been many advances in the efforts to cure patients with hepatitis C virus (HCV). The ultimate goal of these efforts is to develop a combination therapy consisting of only direct-antiviral agents (DAAs). In this paper, we discuss our efforts that led to the identification of a bicyclic template with potent activity against the NS5B polymerase, a critical enzyme on the life cycle of HCV. In continuation of our exploration to improve the stilbene series, the 3,5,6,8-tetrasubstituted quinoline core was identified as replacement of the stilbene moiety. 6-Methoxy-2(1H)-pyridone was identified among several heterocyclic headgroups to have the best potency. Solubility of the template was improved by replacing a planar aryl linker with a saturated pyrrolidine. Profiling of the most promising compounds led to the identification of quinoline 41 (RG7109), which was selected for advancement to clinical development.
Sujet(s)
Antiviraux/pharmacologie , Antienzymes/pharmacologie , Hepacivirus/effets des médicaments et des substances chimiques , Quinoléines/pharmacologie , Sulfonamides/pharmacologie , Protéines virales non structurales/antagonistes et inhibiteurs , Animaux , Antiviraux/composition chimique , Antiviraux/pharmacocinétique , Chiens , Découverte de médicament , Antienzymes/composition chimique , Antienzymes/pharmacocinétique , Hepacivirus/enzymologie , Humains , Modèles moléculaires , Quinoléines/composition chimique , Quinoléines/pharmacocinétique , Rats , Sulfonamides/composition chimique , Sulfonamides/pharmacocinétiqueRÉSUMÉ
Hepatitis C virus (HCV) is a major global public health problem. While the current standard of care, a direct-acting antiviral (DAA) protease inhibitor taken in combination with pegylated interferon and ribavirin, represents a major advancement in recent years, an unmet medical need still exists for treatment modalities that improve upon both efficacy and tolerability. Toward those ends, much effort has continued to focus on the discovery of new DAAs, with the ultimate goal to provide interferon-free combinations. The RNA-dependent RNA polymerase enzyme NS5B represents one such DAA therapeutic target for inhibition that has attracted much interest over the past decade. Herein, we report the discovery and optimization of a novel series of inhibitors of HCV NS5B, through the use of structure-based design applied to a fragment-derived starting point. Issues of potency, pharmacokinetics, and early safety were addressed in order to provide a clinical candidate in fluoropyridone 19.
Sujet(s)
Antiviraux/pharmacologie , Antienzymes/pharmacologie , RNA replicase/antagonistes et inhibiteurs , Protéines virales non structurales/antagonistes et inhibiteurs , Animaux , Antiviraux/composition chimique , Antiviraux/pharmacocinétique , Aire sous la courbe , Lignée cellulaire tumorale , Chiens , Découverte de médicament/méthodes , Antienzymes/composition chimique , Antienzymes/pharmacocinétique , Hepacivirus/effets des médicaments et des substances chimiques , Hepacivirus/enzymologie , Hepacivirus/physiologie , Hépatite C/prévention et contrôle , Hépatite C/virologie , Interactions hôte-pathogène/génétique , Humains , Modèles moléculaires , Thérapie moléculaire ciblée/méthodes , Liaison aux protéines , Structure tertiaire des protéines , Pyridones/synthèse chimique , Pyridones/pharmacocinétique , Pyridones/pharmacologie , RNA replicase/composition chimique , RNA replicase/métabolisme , Rats , Relation structure-activité , Protéines virales non structurales/composition chimique , Protéines virales non structurales/métabolismeRÉSUMÉ
BACKGROUND & AIMS: The combination of a hepatitis C virus (HCV) protease inhibitor, peginterferon, and ribavirin is the standard of care for patients with HCV genotype 1 infection. We report the efficacy and safety of response-guided therapy with danoprevir (a potent second-generation protease inhibitor), peginterferon alfa-2a (40 KD), and ribavirin in these patients. METHODS: Treatment-naïve patients (N = 237) were randomly assigned to groups given 12 weeks of danoprevir (300 mg every 8 hours; 600 mg every 12 hours, and 900 mg every 12 hours) or placebo plus peginterferon alfa-2a and ribavirin, followed by peginterferon alfa-2a and ribavirin. Patients given danoprevir who had an extended rapid virologic response (eRVR4-20: HCV RNA <15 IU/mL during weeks 4-20) stopped therapy at week 24; those without an eRVR4-20 continued therapy to 48 weeks. Patients who were given placebo received 48 weeks of peginterferon alfa-2a and ribavirin. The primary efficacy end point was sustained virologic response (SVR: HCV RNA <15 IU/mL after 24 weeks without treatment). RESULTS: Rates of SVR were higher among patients given danoprevir 300 mg (68%), 600 mg (85%), and 900 mg (76%) than placebo (42%) (95% confidence interval: 26%-59%). Seventy-nine percent of patients given danoprevir 600 mg had an eRVR4-20; among these, 96% had an SVR. Serious adverse events were reported in 7% to 8% of patients given danoprevir and 19% given placebo. Four patients given danoprevir (1 patient in the 600-mg group and 3 in the 900-mg group) had reversible, grade 4 increases in alanine aminotransferase, which led to early discontinuation of the 900-mg arm of the study. CONCLUSIONS: The combination of danoprevir, peginterferon alfa-2a, and ribavirin leads to high rates of SVR in patients with HCV genotype 1 infection, but high doses of danoprevir can lead to grade 4 increases in alanine aminotransferase. Studies of lower doses of danoprevir with ritonavir, to reduce overall danoprevir exposure while maintaining potent antiviral activity, are underway; Clinicaltrials.gov number, NCT00963885.
Sujet(s)
Antiviraux/administration et posologie , Hepacivirus/classification , Hépatite C/traitement médicamenteux , Interféron alpha/administration et posologie , Lactames/administration et posologie , Polyéthylène glycols/administration et posologie , Ribavirine/administration et posologie , Sulfonamides/administration et posologie , Adulte , Cyclopropanes , Association de médicaments , Composés époxy , Femelle , Génotype , Hepacivirus/génétique , Hépatite C/virologie , Humains , Isoindoles , Lactames macrocycliques , Mâle , Adulte d'âge moyen , Proline/analogues et dérivés , Pyridines , Protéines recombinantes/administration et posologieRÉSUMÉ
This analysis was conducted to determine whether the hepatitis C virus (HCV) viral kinetics (VK) model can predict viral load (VL) decreases for nonnucleoside polymerase inhibitors (NNPolIs) and protease inhibitors (PIs) after 3-day monotherapy studies of patients infected with genotype 1 chronic HCV. This analysis includes data for 8 NNPolIs and 14 PIs, including VL decreases from 3-day monotherapy, total plasma trough concentrations on day 3 (C(min)), replicon data (50% effective concentration [EC(50)] and protein-shifted EC(50) [EC(50,PS)]), and for PIs, liver-to-plasma ratios (LPRs) measured in vivo in preclinical species. VK model simulations suggested that achieving additional log(10) VL decreases greater than one required 10-fold increases in the C(min). NNPolI and PI data further supported this result. The VK model was successfully used to predict VL decreases in 3-day monotherapy for NNPolIs based on the EC(50,PS) and the day 3 C(min). For PIs, however, predicting VL decreases using the same model and the EC(50,PS) and day 3 C(min) was not successful; a model including LPR values and the EC(50) instead of the EC(50,PS) provided a better prediction of VL decrease. These results are useful for designing phase 1 monotherapy studies for NNPolIs and PIs by clarifying factors driving VL decreases, such as the day 3 C(min) and the EC(50,PS) for NNPolIs or the EC(50) and LPR for PIs. This work provides a framework for understanding the pharmacokinetic/pharmacodynamic relationship for other HCV drug classes. The availability of mechanistic data on processes driving the target concentration, such as liver uptake transporters, should help to improve the predictive power of the approach.
Sujet(s)
Antiviraux/pharmacocinétique , Hepacivirus/effets des médicaments et des substances chimiques , Inhibiteurs de protéases/pharmacocinétique , Antiviraux/pharmacologie , Humains , Modèles théoriques , Inhibiteurs de protéases/pharmacologieRÉSUMÉ
INTRODUCTION: RG7128 (prodrug of PSI-6130) shows potent antiviral efficacy in patients infected with hepatitis C virus (HCV) genotypes 1, 2, or 3, with mean viral load decreases of 2.7 and 5 log(10) IU/mL, respectively, associated with 1500-mg doses twice daily after monotherapy for 2 weeks and with 1000-mg and 1500-mg doses twice daily after treatment in combination with the standard of care (SOC) for 4 weeks. RESULTS: From 32 patients treated with RG7128 monotherapy for 2 weeks, marginal viral load rebound was observed in 3 HCV genotype 1-infected patients, whereas partial response was observed in 2 genotype 1-infected patients. From 85 patients receiving RG7128 in combination with SOC, 1 HCV genotype 1-infected patient experienced a viral rebound, and 2 genotype 3-infected patients experienced a transient rebound. Five genotype 1-infected patients had an HCV load of >1000 IU/mL at the end of 4-week treatment. No viral resistance was observed, per NS5B sequencing and phenotypic studies. PSI-6130 resistance substitution S282T needs to be present at levels of ≥90% within a patient's quasispecies to confer low-level resistance. No evidence of S282T was found by population or clonal sequence analyses. CONCLUSIONS: The requirement for a predominant S282T mutant quasispecies, its low replication capacity, and the low-level resistance it confers probably contribute to the lack of RG7128 resistance observed in HCV-infected patients.
Sujet(s)
Antiviraux/usage thérapeutique , Désoxycytidine/analogues et dérivés , Hepacivirus/effets des médicaments et des substances chimiques , Hépatite C chronique/traitement médicamenteux , Interféron alpha/usage thérapeutique , Polyéthylène glycols/usage thérapeutique , Ribavirine/usage thérapeutique , Substitution d'acide aminé , Antiviraux/pharmacologie , Désoxycytidine/pharmacologie , Désoxycytidine/usage thérapeutique , Résistance virale aux médicaments/génétique , Association de médicaments , Hepacivirus/génétique , Hepacivirus/physiologie , Hépatite C chronique/virologie , Humains , Interféron alpha-2 , Nouvelle-Zélande , Protéines recombinantes , Sélection génétique , États-Unis , Charge virale , Protéines virales non structurales/génétique , Réplication virale/effets des médicaments et des substances chimiquesRÉSUMÉ
Benzothiazine-substituted tetramic acids were discovered as highly potent non-nucleoside inhibitors of HCV NS5B polymerase. X-ray crystallography studies confirmed the binding mode of these inhibitors with HCV NS5B polymerase. Rational optimization of time dependent inactivation of CYP 3A4 and clearance was accomplished by incorporation of electron-withdrawing groups to the benzothiazine core.
Sujet(s)
Antiviraux/synthèse chimique , Hepacivirus/effets des médicaments et des substances chimiques , Pyrrolidones/composition chimique , Thiazines/composition chimique , Protéines virales non structurales/antagonistes et inhibiteurs , Animaux , Antiviraux/composition chimique , Antiviraux/pharmacocinétique , Sites de fixation , Cristallographie aux rayons X , Pyrrolidones/synthèse chimique , Pyrrolidones/pharmacocinétique , Rats , Relation structure-activité , Protéines virales non structurales/métabolismeRÉSUMÉ
A series of benzo[d]isothiazole-1,1-dioxides were designed and evaluated as inhibitors of HCV polymerase NS5B. Structure-based design led to the incorporation of a high affinity methyl sulfonamide group. Structure-activity relationship (SAR) studies of this series revealed analogues with submicromolar potencies in the HCV replicon assay and moderate pharmacokinetic properties. SAR studies combined with structure based drug design focused on the sulfonamide region led to a novel and potent cyclic analogue.