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461

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Higher Sustained Virologic Response (SVR-12) Achievable in Liver Transplant (LT) Recipients with Hepatitis C (HCV) Treated with Protease Inhibitor (PI) Triple Therapy(TT)

R. Todd Stravitz1, Josh Levitsky2, Jennifer L. Dodge3, Varun Saxena3, James R. Burton5, Elizabeth C. Verna4, Jacqueline G. O'Leary6, Neehar D. Parikh2, Gregory T. Everson5, Robert S. Brown4, James Trotter6, Norah Terrault3;
1Virginia Commonwealth University, Richmond, VA; 2Northwestern University, Chicago, IL; 3University of California at San Francisco, San Francisco, CA; 4Columbia University, New York, NY; 5University of Colorado, Aurora, CO; 6Baylor University, Dallas, TX

Background. HCV recurrence after LT is associated with decreased patient and graft survival, and responds poorly to PEG-interferon/ribavirin (P/R) therapy (30% SVR). The addition of an NS3/4A PI to P/R significantly increases SVR in non-LT patients, but SVR12 rates in LT recipients are unknown. Moreover, the safety and tolerability of TT remain major concerns. Aim. To evaluate safety and efficacy (SVR12) in LT recipients treated with TT. Methods. A total of 122 LT recipients with HCV from 6 US centers (57% GT1a, mean age 58y, 75% male, 45% advanced fibrosis [F3/4], 51% previously treated after LT [48% prior null responders]) were treated at a mean of 3.6y post-LT. Cyclosporine, tacrolimus, or other immunosuppressants were used in 59%, 30%, and 11%, respectively; 76% were also on mycophenolate. A P/R lead-in was used in 97% before the addition of telaprevir (89%) or boceprevir (11%). Efficacy analyses were limited to the 50 patients in whom P/R lead-in was <90d and were eligible to complete ≥51 weeks of TT and follow-up. Safety data were collected on all patients. Results. 36 patients (72%) had end-of-treatment response (EOTR), and 28 (62%; 95% CI 47-76) SVR12. Of 31 (63%) patients achieving eRVR, 94% achieved EOTR and 89% SVR 12, but in those without eRVR, EOTR and SVR 12 were only 39% and 28%, respectively (P<0.001). In patients who received ≤36 wks of TT, SVR12 was achieved in 0/8 without eRVR but 3/4 with eRVR (P=0.02). In patients who received >36 wks of TT, SVR12 was achieved in 5/10 without eRVR but 20/22 with eRVR (P=0.02). 90% of patients with EOTR achieved SVR12, yielding a relapse rate of 9.7%, all within 4 wks post-TT. Adverse events (AEs) led to interruption of TT in 30% and complete discontinuation in 20%. Treated rejection occurred in 3.3% at a median of 168d of TT, but resulted in no graft losses. Maximum serum creatinine increased a median 0.4 (range 0.1-2.5) mg/dl on TT. Despite P/R dose reductions in 41%/82% of patients, erythropoietin was used in 82%, and 52% required a median of 4 units of blood in the first 16 weeks of TT. 7 (5.7%) patients died, 5 of liver-related complications (4 had F3/4 and a mean MELD of 14 at the start of TT), at a median of 234d (range 22-336d) after starting the PI. Conclusions. In LT recipients with recurrent HCV, PI-TT increases SVR12 rates ∼2-fold compared to historical rates with P/R. Relapse after EOTR is uncommon, and occurs early. Duration of TT <36 weeks adversely affects SVR12, especially in those without eRVR, supporting treatment for a full 48 wks. Finally, the incidence and severity of AEs, particularly anemia and renal dysfunction, are considerably higher than in the non-LT population.

Disclosures:

R. Todd Stravitz - Grant/Research Support: Exalenz Biosciences, LTD

Josh Levitsky - Grant/Research Support: Salix, Novartis; Speaking and Teaching: Gilead, Salix, Novartis

James R. Burton - Grant/Research Support: Vertex pharaceuticals, Abbvie pharmaceuticals, Gilead pharmaceuticals

Jacqueline G. O'Leary- Consulting: Vertex, Gilead

Gregory T. Everson - Advisory Committees or Review Panels: Roche/Genentech, Merck, HepC Connection, Roche/Genentech, Merck, HepC Connection; Board Membership: HepQuant LLC, PSC Partners, HepQuant LLC, PSC Partners; Consulting: Roche/Genentech, BMS, Gilead, Roche/Genentech, Bristol-Myers Squibb, Abbott; Grant/Research Support: Roche/Genentech, Pharmassett, Vertex, GSK, Schering-Plough, Bristol-Myers Squibb, Tibotec, GlobeImmune, Pfizer, Abbott, Conatus, Zymogenetics, PSC Partners, Roche/Genentech, Pharmassett, Vertex, GSK, Schering-Plough, Tibotec, GlobeImmune, Pfizer, Gilead, Conatus, Zymogenetics, PSC Partners, Abbott; Management Position: HepQuant LLC, HepQuant LLC; Patent Held/Filed: Univ of Colorado, Univ of Colorado

Robert S. Brown - Consulting: Salix, Janssen, Vertex; Grant/Research Support: Gilead, Merck, Vertex, AbbVie, Salix, Janssen, BI; Speaking and Teaching: Genentech, Gilead, Merck

James Trotter - Speaking and Teaching: Salix, Novartis

Norah Terrault - Advisory Committees or Review Panels: Eisai, Biotest; Consulting: BMS; Grant/Research Support: Eisai, Biotest, Vertex, Gilead, AbbVie, Novartis

The following people have nothing to disclose: Jennifer L. Dodge, Varun Saxena, Elizabeth C. Verna, Neehar D. Parikh

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Association of gamma-glutamyl transferase levels and hepatocellular carcinoma development in non-cirrhotic patients with hepatitis C virus eradication

Chia-Yen Dai, Chung-Feng Huang, Jee-Fu Huang, Ming-Lung Yu, Wan-Long Chuang; Hepatobiliary Division,
Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, Kaohsiung, Taiwan

Background/Aim Serum gamma-glutamyl transferase (r-GT) levels were associated with liver disease severity. We aimed to explore the association of r-GT and HCV-related HCC development in patients with a sustained virological response (SVR). Methods Clinical parameters including r-GT levels of 856 patients who achieved an SVR were evaluated from 2002 to 2010. Results Thirty-three patients (3.9 %) developed HCC within a median follow-up period of 44.2 months (range 9-91 months). Cox regression analysis revealed that the strongest factor predictive of HCC occurrence was liver cirrhosis (hazard ratio [HR] 5.49, 95% confidence intervals [CI.] 1.74-8.37, P<0.001), followed by age (HR 1.06, 95% CI. 1.02-1.06, P=0.005) and r-GT levels (HR 1.008, 95% CI. 1.004-1.013, P=0.001). The r-GT levels did not differ between cirrhotic patients with or without HCC (77.7+64.7 u/L vs. 75.0+67.8 U/L, P=0.93), and the incidence of HCC did not differ between patients with high or low r-GT levels (log-rank test P=0.49). On the contrary, the r-GT levels were significantly higher in non-cirrhotic patients with HCC development than those without (100.3+79.2 u/L vs. 61.8+54.8 U/L, P=0.03), and the incidence of HCC was significantly higher in those with high r-GT levels as compared with those without (log-rank test P=0.004). Cox regression analysis revealed that the strongest factor associated with HCC development in non-cirrhotic patients was high r-GT levels (HR 5.28, 95% CI. 1.73-16.17, P=0.004), followed by male gender (HR 4.69, 95% CI. 1.26-17.38, P=0.02), age (HR 4.24, 95% CI. 1.43-12.57, P=0.009) and hemoglobin concentrations (HR 0.64, 95% CI. 0.47-0.88, P=0.005). Conclusions HCC remains a threat in non-cirrhotic patients with an SVR. Serum r-GT levels helped to identify the potential patients at high risk.

Kaplan-Meier analysis of the time to HCC development in non-cirrhotic patients with low or high serum r-GT levels

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Disclosures:

Ming-Lung Yu - Advisory Committees or Review Panels: ABBOTT, MSD; Grant/Research Support: ABBOTT, ROCHE, MSD; Speaking and Teaching: ABBOTT, ROCHE, MSD, GILEAD, BMS, GSK

Wan-Long Chuang - Advisory Committees or Review Panels: Gilead, Roche, Norvatis; Speaking and Teaching: BMS

The following people have nothing to disclose: Chia-Yen Dai, Chung-Feng Huang, Jee-Fu Huang

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Asunaprevir Pharmacokinetics and Safety in Subjects With Impaired Renal Function

Tushar Garimella1, Bing He1, Wen-Lin Luo2, Elizabeth Colston1, Kurt Zhu1, Hamza Kandoussi2, Thomas C. Marbury3, Harry W. Alcorn4, William B. Smith5, Timothy Eley1;
1Research and Development, Bristol-Myers Squibb, Hopewell, NJ; 2Research and Development, Bristol-Myers Squibb, Lawrenceville, NJ; 3Orlando Clinical Research Center, Orlando, FL; 4DaVita Clinical Research, Minneapolis, MN; 5New Orleans Center for Clinical Research, Knoxville, TN

Background Asunaprevir (ASV, formerly BMS-650032) is a selective HCV NS3 protease inhibitor with in vitro activity against genotypes 1, 4, 5 and 6. ASV has been demonstrated to be safe and efficacious as part of multiple (including all-oral) regimens. ASV is primarily excreted via the feces with minimal renal excretion. Study AI447-033 assessed the pharmacokinetics (PK) and safety of the Phase 3 ASV soft capsule in subjects with end-stage renal disease (ESRD) compared with matched healthy controls with normal renal function. Methods A reduced study design was utilized per FDA guidance on assessing renal impairment for drugs primarily eliminated by hepatic metabolism. In this open-label, parallel, multiple dose study, 12 subjects with normal renal function (Group A, creatinine clearance rate of >90 mL/min) and 12 subjects with ESRD (Group B, estimated glomerular filtration rate of <15 mL/min/1.73m2) received ASV 100 mg BID on Days 1-6 and morning dose on Day 7. Blood samples for PK were collected for 12 hours post-dose on Day 1 and for 72 hours post-dose on Day 7. Plasma concentrations were determined using a validated LC/MS/MS method. Noncompartmental PKwere derived. Geometric mean ratios (GMR) and 90% confidence intervals (90%CI) were calculated for ASV Cmax and AUCTAU using an ANCOVA model containing categorical variables for population (Groups A and B) and gender, and continuous covariates for age and weight. Subjects were monitored for adverse events (AEs) throughout the study. Results Twelve subjects (8 males and 4 females) were enrolled in each group and completed the study. The mean age was 53 years (range 40-74) and mean BMI was 27.3 kg/m2 (range 19.3-33.3). All 12 subjects in group B were on hemodialysis. Day 7 geometric mean PK parameters (% CV) are shown in the table. The Group B/Group A GMR (90% CI) for ASV AUCTAU was 0.90 (0.63, 1.28) and for ASV Cmax was 1.29 (0.76, 2.17), supporting the research hypothesis that ASV PK would not be altered in subjects with renal impairment in a clinically significant manner. One subject experienced a serious AE (thrombosis of arteriovenous fistula requiring hospitaliza-tion and treatment) not related to study drug and no subjects were discontinued due to AEs. Conclusions Severe renal impairment had minimal impact on the PK of ASV; data suggest no dose adjustment is needed for ASV in subjects with any level of renal impairment.

Table 1. 
 Geometric Mean (%CV)
ParameterGroup A (Healthy)Group B (ESRD)
Cmax101.3(141)133(60.4)
AUCTAU429.4(65.7)393.2(50.1)
C126.8(66.4)7.59(36.1)

Disclosures:

Tushar Garimella - Employment: Bristol Myers-Squibb; Stock Shareholder: Abbvie

Bing He - Employment: Bristol-Myyers Squibb

Wen-Lin Luo - Employment: BMS

Elizabeth Colston - Employment: Bristol-Myers Squibb

Kurt Zhu - Employment: Bristol-Myers Squibb

Thomas C. Marbury- Employment: Orlando Clinical Research Center

Timothy Eley- Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers Squibb

The following people have nothing to disclose: Hamza Kandoussi, Harry W. Alcorn, William B. Smith

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Lack of Pharmacokinetic Interaction Between HCV Protease Inhibitor MK-5172 and HCV NS5A Inhibitor Daclatasvir in Healthy Volunteers

Wendy W. Yeh1, Iain P. Fraser1, Marc Bifano2, Luzelena Caro1, Jennifer E. Talaty1, Zifang Guo1, Jennifer M. McCarthy1, Stephen P. Youngberg3, Joan R. Butterton1;
1Merck Sharp & Dohme Corp., Whitehouse Station, NJ;2Bristol-Myers Squibb Research and Development, Hopewell, NJ; 3Celerion, Lincoln, NE

Background: MK-5172, a potent, once-daily competitive inhibitor of the hepatitis C virus (HCV) NS3/4A protease, and daclatasvir (DCV), an HCV NS5A replication complex inhibitor with pan-genotypic activity in vitro, are being developed for the treatment of chronic HCV infection. The aim of the present study was to evaluate potential pharmacokinetic interactions as well as safety and tolerability of MK-5172 and daclatasvir co-administration in healthy subjects. Methods: This was a single-center, open-label, fixed-sequence, multiple-dose study in 14 healthy adult male and female volunteers, ages 19-49 years. Since MK-5172 in HCV-infected patients demonstrates ∼2-fold higher exposure compared to healthy subjects, a 200 mg dose of MK-5172 in healthy subjects was used in this study to match the exposure of a 100 mg dose (the intended clinical dose) in HCV-infected patients. In Period 1, subjects received oral doses of 60 mg daclatasvir once daily on Days 1 to 7. Following a 4 day washout, subjects received oral doses of 200 mg MK-5172 once daily on Days 1 to 7 in Period 2. In Period 3, which commenced immediately after Period 2, subjects were co-administered once daily oral doses of 200 mg MK-5172 and 60 mg daclatasvir on Days 1 to 8. Plasma pharmacokinetic samples were obtained for daclatasvir on Day 7 in Period 1 and Day 8 in Period 3, as well as for MK-5172 on Day 7 in Period 2 and Day 8 in Period 3. Safety assessments included electrocardiograms, vital signs, clinical laboratory tests, physical examination, and adverse event monitoring. Results: Co-administration of MK-51 72 with daclatasvir was safe and well-tolerated. Multiple oral doses of MK-51 72 did not meaningfully change the steady-state AUC0-τ, Cmax, or C24h of daclatasvir with MK-5172+DCV/DCV geometric mean ratios (GMRs) [90% confidence intervals (CIs)] of 1.02 [0.93, 1.11], 0.80 [0.74, 0.86], and 1.23 [1.09, 1.38], respectively. Multiple oral doses of daclatasvir did not meaningfully change the steady-state AUC0-τ, Cmax, or C24h of MK-5172 with MK-5172+DCV/MK-5172 GMRs [90% CIs] of 1.12 [0.87, 1.44], 1.11 [0.77, 1.60], and 1.04 [0.97, 1.12], respectively. Conclusions: Co-administration of MK-5172 and daclatasvir in healthy volunteers did not result in clinically significant drug-drug interactions. MK-5172 and DCV were safe and well-tolerated when coadministered. These results suggest that no dose adjustments of MK-5172 or daclatasvir are needed for co-administration of these drugs in interferon-free, combination regimens containing these once-daily direct acting antivirals in HCV-infected patients.

Disclosures:

Wendy W. Yeh - Employment: Merck & Co.

Iain P. Fraser - Employment: Merck & Co.; Stock Shareholder: Merck & Co.

Marc Bifano - Employment: Bristol-Myers Squibb

Luzelena Caro - Employment: Merck & Co., Inc.

Jennifer E. Talaty - Employment: Merck, Sharp, & Dohme

Zifang Guo - Employment: Merck & Co., Inc.

Stephen P. Youngberg - Employment: Celerion, Inc.

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

The following people have nothing to disclose: Jennifer M. McCarthy

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Lack of a Clinically Significant Pharmacokinetic Drug-Drug Interaction between Sofosbuvir and GS-5816 in Healthy Volunteers

Erik Mogalian, Polina German, Diana M. Brainard, John O. Link, John McNally, Lingling Han, Brian P. Kearney;
Gilead Sciences, Inc, Foster City, CA

Background Sofosbuvir (SOF) is a specific nucleotide HCV NS5B polymerase inhibitor. GS-5816 is a second generation HCV NS5A inhibitor with picomolar antiviral activity against genotypes 1-6. The combination of SOF and GS-5816 may constitute a regimen with broad genotypic activity for the treatment of patients with chronic HCV infection. Thus, a drug-drug interaction study between SOF and GS-5816 was conducted in healthy volunteers. Methods In this open-label, fixed-sequence, cross-over, drug-drug interaction study, subjects received a single dose (SD) of SOF 400 mg on Day 1, once-daily doses of GS-5816 150 mg on Days 5-13, and a SD of SOF 400 mg coadministered with GS-5816 150 mg on Day 14. All doses were administered under fed conditions. Safety assessments were performed throughout the study. Geometric means ratios (GMR%: combination vs. alone) and 90% confidence intervals (CIs) for AUCinf and Cmax of SOF and GS-331007 (the predominant circulating nucleoside metabolite of SOF) and AUC-tau, Cmax and Ctau of GS-5816 were estimated using ANOVA. Lack of pharmacokinetic (PK) interaction bounds were set as 70% to 143%. Results All enrolled subjects (N=18) completed the study. Study drugs were generally well tolerated. Three subjects receiving SOF alone, 3 subjects receiving GS-5816 alone, and 4 subjects receiving SOF+GS-5816 experienced a treatment-emergent adverse event (AEs). All AEs were Grade 1 (mild); one AE (constipation) was considered study drug-related. GMR% (90% CI) upon coadministration vs. treatment alone are presented in the table below. SOF plasma exposure increased ∼1.8-2.4-fold when coadministered with GS-5816. The effect of GS-5816 on SOF is likely due to inhibition of intestinal P-gp and/or BCRP, of which SOF is a known substrate. The magnitude of the increase in exposure for SOF was comparable to that seen when SOF was coadministered with the first-generation NS5A inhibitor ledipasvir. For GS-331007, an approximately 35% lower Cmax was observed upon SOF administration with GS-5816. The GMR% (90% CI) for GS-331007 AUC were within the equivalence bounds. Since the decrease in GS-331007 Cmax was modest and the AUC parameters met the equivalence criteria, the effect of GS-5816 on GS-331007 PK was not considered clinically significant. Coadministration of SOF with GS-5816 did not alter GS-5816 PK. Conclusion Coadministration of SOF and GS-5816 was generally well tolerated. Sofosbuvir and GS-5816 may be coadministered without dose adjustment.

Table 2. 
SOF+GS-816/SOF or GS-5816 Alone, GMR% (90% CI)
SOFGS-331007GS-5816
AUCinfCmaxAUCinfCmaxAUCinfCmax
238(215,264)181 (149,219)116(111, 122)64.2 (58.5,70.4)112(108, 116)106(102, 110)

Disclosures:

Erik Mogalian - Employment: Gilead Sciences, Inc; Stock Shareholder: Gilead Sciences, Inc

Polina German - Employment: GIlead Sciences, Inc; Stock Shareholder: GIlead Sciences, Inc

Diana M. Brainard - Employment: Gilead Sciences, Inc.

John O. Link - Employment: Gilead Sciences; Patent Held/Filed: Gilead Sciences; Stock Shareholder: Gilead Sciences, Pfizer

John McNally- Employment: Gilead Sciences, Inc

Brian P. Kearney- Employment: Gilead Sciences

The following people have nothing to disclose: Lingling Han

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Pharmacokinetics, safety, and tolerability of faldaprevir in patients with different levels of renal impairment

Fenglei Huang1, Viktoria Moschetti2, Benjamin Lang3, Atef Halabi4, Marc Petersen-Sylla4, Chan-Loi Yong1, Mabrouk Elgadi5;
1Boehringer Ingelheim Pharmaceuticals Inc., Ridgefield, CT; 2Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim, Germany; 3Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany; 4CRS Clinical Research Services Kiel GmbH, Kiel, Germany; 5Boehringer Ingelheim Ltd, Burlington, ON, Canada

Background: Faldaprevir (FDV) is a potent HCV NS3/4A protease inhibitor. In combination with pegylated interferon alfa-2a and ribavirin (RBV), or with BI 207127 and RBV, FDV has demonstrated high sustained virologic response rates in treatment-naïve patients with chronic HCV genotype 1 (GT1) infection. Here, we have assessed the pharmacokinetics (PK) and safety of a single dose of FDV in subjects with varying levels of renal impairment. Methods: HCV-negative subjects (18-75 years) with renal impairment (mild to severe based on estimated glomerular filtration rate [eGFR]), and healthy controls, were given a single oral dose of FDV 480 mg after a 10-hour overnight fast. PK and safety assessments were performed over a 144-hour period after dosing. Results: 32 subjects (mean age 61.4 years; 21 males) completed the study. Due to mild vomiting events, 8 subjects were excluded from the primary PK analysis. Geometric mean (gMean) peak (Cmax) and total (AUC0 J exposures of FDV were highest in subjects with moderate renal impairment (Table). Compared with subjects with normal renal function, the statistical analysis showed that adjusted gMean ratios (90% CI) for AUC0-∞ were 1.14 (0.42-3.10), 1.78 (0.85-3.73), and 1.69 (0.73-3.91) for subjects with mild, moderate, or severe renal impairment, respectively. The gMean ratios for C were 1.07 (0.35-3.27), 1.76 (0.90-3.44), and 1.21 (0.47-3.10). Median tmax was 4 hours for all groups. There was no difference in the free fraction of FDV (not bound to plasma proteins) for subjects with any renal impairment versus those with normal renal function. Although renal impairment increased peak and total exposure, no correlation between either AUC0-∞ or Cmax and eGFR was found. Overall, 5/8 (63%) subjects with normal renal function and 20/24 (83%) subjects with renal impairment reported adverse events. There were no notable differences in treatment tolerability between subjects with normal renal function and those with renal impairment. Conclusions: Moderate or severe renal impairment can result in a modest increase in exposure to FDV. However, given the wide therapeutic range and tolerability profile observed in the FDV clinical development program, dose adjustment in HCV patients with renal impairment is not warranted.

Table 3. 
 Normal (n=5)Mild (n=4)Moderate(n=7)Severe (n=8)
eGFR, mL/min/1.73m2≥9060-8930-5915-29
Cmax,ng/mL; gMean (gCV [%])3810(80.7)4080 (144)6680 (63.2)4600(135)
AUC0-∞, ng.h/mL; gMean (gCV [%])76500 (95.7)86 900 (89.8)136 000(67.2)129 000 (98.8)
t1/2,h36.135.833.131.8

Disclosures:

Fenglei Huang - Employment: Boehringer Ingelheim Pharmaceuticals, Inc

Viktoria Moschetti - Employment: Boehringer Ingelheim Pharma GmbH&Co. KG

Benjamin Lang - Employment: Boehringer Ingelheim Pharma GmbH & Co. KG

Marc Petersen-Sylla - Employment: CRS-Kiel

Mabrouk Elgadi - Employment: Boehringer Ingelheim

The following people have nothing to disclose: Atef Halabi, Chan-Loi Yong

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The Pharmacokinetics of Ledipasvir, an HCV Specific NS5A Inhibitor in HCV-Uninfected Subjects with Moderate and Severe Hepatic Impairment

Polina German1, Anita Mathias1, Jenny C. Yang1, Lindsay McNair1, Gong Shen1, Mona Vimal1, Edward J. Gane2, William B. Smith4, Gernot K. Klein6, Thomas C. Marbury5, Eric Lawitz3;
1GIlead Sciences, Inc, Foster City, CA, CA; 2Auckland Clinical Studies, Auckland, New Zealand; 3Alamo Medical Research, San Antonio, TX; 4New Orleans Center for Clinical Resarch, Knoxville, TN; 5Orlando Clinical Research Center, Orlando, FL; 6APEX Research, Munchen, Germany

Ledipasvir (LDV), a potent HCV NS5A inhibitor, is in Phase 3 clinical development for the treatment of chronic HCV infection as a fixed-dose combination tablet with sofosbuvir. LDV is primarily eliminated in the feces as an unchanged parent drug (∼ 70% of the dose); ∼1 % of the LDV dose is excreted in the urine as metabolites. Since many HCV-infected patients may develop impaired hepatic function during the natural history of the disease, this study evaluated the short-term safety and pharmacokinetics (PK) of LDV in subjects with moderate or severe hepatic impairment (HI) versus control subjects with normal hepatic function (NF) to inform dosing recommendations for LDV in this population. Methods Subjects with stable moderate hepatic impairment (N=10) Child-Pugh-Turcotte (CPT) Classification B (score 7- 9) and healthy control subjects with normal hepatic function, matched for age (±10 years), gender, and BMI (±15%) received LDV 30 mg+GS-9451 200 mg daily (N=10) for 12 days each with food. Subjects with stable severe HI (N = 10) CPT C (score 10-15) and matched controls received a single dose (SD) of LDV 90 mg with food. All treatments were followed by intensive pharmacokinetic (PK) sampling. Safety assessments were performed throughout the study. Geometric mean ratios (GMRs: HI:NF) and 90% confidence intervals (CIs) for LDV AUC (tau/inf), Cmax and Ctau (moderate HI only) were calculated using ANOVA model with an exposure increase of at least 100% being considered as clinically relevant. Results All enrolled subjects (N=10/group) completed the study; no subject discontinued due to an adverse event (AE). One moderate HI subject was excluded from analysis due to a major protocol deviation (disallowed medication). All treatment-emergent AEs were Grade 1 (mild), except for one Grade 2 (moderate) AE (headache: severe HI subject). LDV plasma exposures were similar in subjects with moderate HI and controls; LDV Cmax was modestly lower but AUC remained comparable in subjects with severe HI and normal hepatic function. Conclusions: LDV administration was safe and well tolerated. No clinically relevant changes in overall LDV plasma exposures were observed in subjects with moderate or severe hepatic impairment relative to subjects with normal hepatic function. LDV dose adjustment is therefore not required in patients with chronic HCV infection with mild, moderate or severe hepatic impairment.

Table 4. 
  1. * AUCtau for moderate HI; AUCinf for severe HI; Ctau: not reported for SD

LDV PK Parameter*Moderate HI vs. NF (N=9) GMR% [90 % CI])Severe HI vs. NF (N=10) GMR% [90 % CI])
AUC100 (63.4, 158)108(70.1, 167)
Cmax90.8(59.3, 139)64.6 (46.5, 89.8)
Ctau105(62.2, 179)NA

Disclosures:

Polina German - Employment: GIlead Sciences, Inc; Stock Shareholder: GIlead Sciences, Inc

Anita Mathias - Employment: Gilead Sciences Inc., Jenny C. Yang - Employment: Gilead Sciences Lindsay McNair - Independent Contractor: Gilead

Edward J. Gane - Advisory Committees or Review Panels: Roche, AbbVie, Novar-tis, Tibotec, Gilead Sciences, Janssen Cilag, Vertex, Achillion; Speaking and Teaching: Novartis, Gilead Sciences, Roche

Thomas C. Marbury- Employment: Orlando Clinical Research Center

Eric Lawitz - Advisory Committees or Review Panels: AbbVie, Achillion Pharmaceuticals, BioCryst, Biotica, Enanta, Idenix Pharmaceuticals, Janssen, Merck & Co, Novartis, Santaris Pharmaceuticals, Theravance, Vertex Pharmaceuticals; Grant/Research Support: AbbVie, Achillion Pharmaceuticals, Boehringer Ingelheim, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Idenix Pharmaceuticals, Intercept Pharmaceuticals, Janssen, Merck & Co, Novartis, Presidio, Roche, Santaris Pharmaceuticals, Vertex Pharmaceuticals ; Speaking and Teaching: Gilead, Kadmon, Merck, Vertex

The following people have nothing to disclose: Gong Shen, Mona Vimal, William B. Smith, Gernot K. Klein

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An HCV-HCC susceptibility gene MICA found in GWAS was effectively induced by HDAC inhibitors

Kaku Goto, Ryosuke Muroyama, Wenwen Li, Norie Kowatari, Ryo Nakagawa, Naoya Kato;
The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

Background and aims: We reported that MHC class I polypeptide-related sequence A (MICA) was a genetic susceptibility factor for hepatitis C virus (HCV)-induced hepatocellular carcinoma (HCC) in a genome-wide association study (Kumar V et al., Nat Genet 2011). The risk of HCC development was elevated by decreased MICA expression in HCV-infected patients, indicating anti-hepatocarcinogenic effects of MICA upregulation. Hence we aimed to find inducers of MICA expression using a reporter screen system. Methods: Human hepatoma cell lines and the JFH1 infection system were used. Intracellular mRNA levels for individual genes were measured by qRT-PCR. Transcriptional activities of MICA promoter was monitored via luciferase activities of reporter plasmids. Stable transformant cells were established by the selection with puromycin. Cell viability was assessed by tetrazolium salt assay. Results: Cotreatment with valproic acid (VPA) and hydroxyurea (HU), reported inducers of MICA in leukemic cell lines, elevated MICA mRNA levels in hepatoma cells. Then we generated luciferase reporters harboring MICA promoter sequences and their activities were enhanced by VPA and HU. Subsequently stable transformant cells carrying the reporters were selected by puromycin, which also positively responded to the VPA/HU cotreatment. This reporter cell system has so far detected increased MICA transcriptional activities consistent with the mRNA level augmentation by several compounds including short chain fatty acids and histone deacetylase inhibitors in a drug library at noncytotoxic doses. Furthermore certain MICA-inducing drugs identified here even demonstrated antiviral activities in the JFH1 infection system. Conclusions: Drugs found in our reporter system induced MICA expression effectively indeed, and would serve to devise anti-HCC strategies in HCV infection.

Disclosures:

The following people have nothing to disclose: Kaku Goto, Ryosuke Muroyama, Wenwen Li, Norie Kowatari, Ryo Nakagawa, Naoya Kato

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Lack of a Clinically Important Pharmacokinetic Interaction between Norgestimate/Ethinyl Estradiol and Sofos-buvir (SOF) or Ledipasvir (LDV) in HCV-Uninfected Female Subjects

Polina German1, Lisa Moorehead4, Phil S. Pang2, Mona Vimal3, Anita Mathias1;
1Clinical Pharmacology, Gilead Sciences, Inc, Foster City, CA; 2Clinical Research, Gilead Sciences, Inc, Foster City, CA; 3Clinical Operations, Gilead Sciences, Inc, Foster City, CA; 4Biostatistics, Gilead Sciences, Inc, Foster City, CA

SOF, an HCV-specific NS5B nucleotide inhibitor with a broad genotype (GT) coverage and LDV, an HCV NS5A inhibitor are in development for the treatment of chronic HCV infection. Since HCV-infected female patients on oral hormonal contraceptives (OC) may be treated with SOF or SOF/LDV fixed dose combination (FDC), this study evaluated a potential for a drug-drug interaction between SOF or LDV and norgestimate/ethinyl estradiol (NGM/EE, Ortho Tri-Cyclen Lo®), a representative hormonal oral contraceptive (OC). Methods This was an open-label, fixed-sequence, Phase 1 study. Subjects not using NGM/EE were enrolled into Part A (lead-in) and received NGM/EE for 1 menstrual cycle before enrolling into Part B (main study). Subjects on NGM/EE could enroll into Part B directly. In Part B, subjects received NGM/EE for 3 sequential cycles. NGM/EE was administered alone (1st cycle), followed by coadministration with SOF for 7 days (Days 8-14; 2nd cycle) or with LDV for 14 days (Days 1-14; 3rd cycle). Safety assessments were conducted throughout the study. NGM, norelgestromin (NGMN; active metabolite), norgestrel (NG; active metabolite), EE, SOF and GS-331007 (predominant circulating nucleoside metabolite of SOF), and LDV were analyzed on Day 14 of each respective cycle. Geometric least squares mean ratios (GLSMR) and 90% confidence intervals (CIs) for AUCtau, Cmax and Ctau were estimated using ANOVA with PK alteration bounds of 70-143%. FSH (Day 14) and LH (Day 14), and progesterone (Day 21) were assessed in all cycles. Results All enrolled subjects (N=15) completed Part B. Study treatments were well tolerated. Nausea and headache were the most frequently reported AEs. All treatment-emergent AEs were mild (Grade 1) or moderate (Grade 2). Small increases in EE Cmax (∼40%) with LDV or NG AUCtau (∼19%) and Ctau (∼23%) with SOF were noted. No other alterations in NGM/EE PK were observed. SOF, GS-331007 and LDV PK were similar to historical data. FSH, LH and progesterone values were similar in all cycles. Conclusion Coadministration of SOF or LDV with NGM/EE was safe and well tolerated. Based on these results, no loss in contraceptive efficacy is expected upon administration of combined oral contraceptives containing ethinyl estradiol and norgestimate with SOF or SOF/LDV FDC. Accordingly, the use of OC with SOF or SOF/LDV FDC is permitted.

Table 5. 
Analyte (N=15)SOF+NGM/EE vs. NGM/EE GLSMRs% (90% CI)LDV+NGM/EE vs NGM/EE GLSMRs%(90% CI)
AUCtauCmaxCtauAUCtauCmaxCtau
  1. *NGM was BLQ for all subjects at most timepoints

NGMN106(92.3, 121)107(93.6, 122)107(89.0, 128)103(90.1, 118)102(89.2, 116)109(91.2, 131)
NO119(98.4, 145)118(99.2, 141)123(99.8, 151)99.0(81.5, 120)103(86.5, 123)100(81.5, 123)
EE109 (94.0, 126)115(96.6, 136)98.9 (79.7, 123)120(104, 139)140(118, 166)98.3 (79.2, 122)

Disclosures:

Polina German - Employment: GIlead Sciences, Inc; Stock Shareholder: GIlead Sciences, Inc

Lisa Moorehead - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences

Phil S. Pang - Employment: Gilead Sciences

Anita Mathias - Employment: Gilead Sciences Inc.,

The following people have nothing to disclose: Mona Vimal

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Deep Sequencing of HCV NS5A From a 3-Day Study of GS-5816 Monotherapy Confirms the Potency of GS-5816 Against Pre-Existing Genotype 1-3 NS5A Resistance-Associated Variants

Christy Hebner1, Viktoria Gontcharova1, Ramakrishna K. Cho-davarapu1, Maribel Rodriguez-Torres2, Eric Lawitz3, Chris Yang1, John McNally1, John O. Link1, Hongmei Mo1;
1Clinical Virology, Gilead Sciences, Inc., Foster City, CA; 2Fundacion De Investiga-cion, San Juan; 3The Liver Institute, University of Texas Health Science Center, San Antonio, TX

Background: GS-5816 is a second generation HCV NS5A inhibitor with picomolar antiviral activity against HCV genotypes 1 -6 that also maintains antiviral activity against signature NS5A resistance associated variants. Here we report NS5A deep sequencing analyses of patient HCV samples obtained from a 3-day monotherapy study of GS-5816 in HCV-infected subjects. Methods: Treatment naïve patients with chronic HCV infection were administered GS-581 6 for 3 days at doses ranging from 5mg-150mg. Pretreatment, on-treatment, and post-treatment plasma samples from GT 1,2, and 3 subjects were analyzed for NS5A RAVs by deep sequencing analysis with a 1% assay cutoff. The presence of NS5A RAVs at amino acids 28, 30, 31, 58 and 93 were examined for GT 1a, 2, and 3 samples and NS5A amino acids 31 and 93 for GT 1 b. Results: The NS5A RAVs M28T, Q30H/R, L31M/V, and Y93C/H/R were detected at baseline in 10/32 (31.3%) sequenced GT 1 a infected subjects. Despite this high rate of detectable RAVs, similar viral load decreases were observed in subjects with detectable baseline RAVs versus those with no detectable RAVs. One GT 1 b infected subject had Y93H at baseline that did not impact response to GS-5816 150mg. Four of seven GT 2 infected subjects had detectable L31M at baseline. No significant differences in maximum viral load reductions between subjects with L31 or M31 were observed. In GT 3 infected subjects, the A30K and Y93H RAVs were observed in 1/10 and 2/10 subjects, respectively. One GT 3 infected subject with Y93H (16% of population) had a 2.7 log 10 reduction in HCV RNA with administration of GS-5816 150 mg. The single GT 3 infected subject with A30K had a 2.9 log10 decrease in HCV RNA. Overall, NS5A RAVs were detected at varying prevalence, with frequencies varying from 1-99% as a proportion of the HCV quasispecies. Fourteen-day post-treatment analyses demonstrated complex mixtures of NS5A RAVs including M28T, Q30R/H, L31M/V, H58D, and Y93H/N/S among GT 1a subjects. In GT 1b and GT 2b subjects, L31V/M and Y93H were commonly observed while Y93H/N was the most common RAV in GT 3 subjects following 3-day GS-5816 treatment. Conclusions: The NS5A inhibitor GS-5816 demonstrated potent antiviral activity in GT 1,2, and 3 HCV-infected subjects despite the presence of NS5A RAVs at baseline. The patterns of NS5A RAVs observed after GS-5816 monotherapy were GT/subtype dependent.

Disclosures:

Christy Hebner - Employment: Gilead Sciences, Inc.

Ramakrishna K. Chodavarapu - Employment: Gilead Sciences, Inc

Maribel Rodriguez-Torres - Consulting: Hoffman La Roche, Abbott Labs, Pharmasset, Akros, Bristol-Myers Squibb, Merck, Vertex, Inhibitex, Genentech, Janssen R&D Ireland, Santaris; Grant/Research Support: Anadys, Novartis, Hoffman-LaRoche, Glaxo Smith Kline, Inhibitex, Bristol-Myers Squibb, Vertex, Idera, Pharmasset, Sanofi-Aventis, Merck, Abbott, Pfizer, Human Genome Sciences, Gilead, Johnson & Johnson, Zymogenetics, AKROS, Scynexis, Santaris, Boehringher, Idenix, Genentech, Beckman Coulter, Mochida Pharmaceutical

Eric Lawitz - Advisory Committees or Review Panels: AbbVie, Achillion Pharmaceuticals, BioCryst, Biotica, Enanta, Idenix Pharmaceuticals, Janssen, Merck & Co, Novartis, Santaris Pharmaceuticals, Theravance, Vertex Pharmaceuticals; Grant/Research Support: AbbVie, Achillion Pharmaceuticals, Boehringer Ingel-heim, Bristol-Myers Squibb, Gilead Sciences, GlaxoSmithKline, Idenix Pharmaceuticals, Intercept Pharmaceuticals, Janssen, Merck & Co, Novartis, Presidio, Roche, Santaris Pharmaceuticals, Vertex Pharmaceuticals ; Speaking and Teaching: Gilead, Kadmon, Merck, Vertex

Chris Yang - Employment: Gilead Sciences; Stock Shareholder: Gilead Sciences John McNally- Employment: Gilead Sciences, Inc

John O. Link - Employment: Gilead Sciences; Patent Held/Filed: Gilead Sciences; Stock Shareholder: Gilead Sciences, Pfizer

Hongmei Mo - Employment: Gilead Science Inc

The following people have nothing to disclose: Viktoria Gontcharova

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Genetic variants of the Niemann-Pick C1-like 1 cholesterol absorption receptor and response to antiviral therapy in HCV patients

Joaquin Cabezas1,2, Emilio Fábrega1,2, Ignacio Varela3, Jose Luis Fernandez Luna2,4, Ana Fontalbo2,4, Juan Antonio Gomez Gerique6, Jose A. Del Campo5, Angela Rojas5, Angela Puente1,2, Maria Teresa Arias1,2, Marta García-Valdecasas5, Manuel Romero-Gomez5, Javier Crespo1,2;
1Gastroenterology and Hepatology Department, Marques de Valdecilla University Hospital, Santander, Spain; 2Marques de Valdecilla Fundation (IFIMAV), Santander, Spain; 3Biomedicin and Biotecnology Cantabria Institut. Massive Sequencing Service, Santander, Spain; 4Molecular Genetics Unit, Marques de Valdecilla Universitary Hospital, Santander, Spain; 5UCM Digestive Diseases and CIBEREHD, Valme University Hospital. University of Sevilla, Sevilla, Spain; 6Clinical Analisys Service, Marques de Valdecilla University Hospital, Santander, Spain

Background: A recently discovered novel surface receptor involved in HCV entry, the Niemann-Pick C1-like 1 cholesterol (NPC1L1), is an HCV entry factor amendable to therapeutic intervention. Previously, DNA sequencing studies revealed that nonsynonymous variants in NPC1L1 are collectively common in general population. The aim of the present study was to elucidate whether genetic variants of the NPC1L1 are linked to the antiviral response in a group of patients with chronic hepatitis C (CHC). Methods: We included 38 patients with CHC genotype 1 treated with pegylated interferon alpha2 and ribavirin (20 patients with SVR and 18 null responders). The whole coding sequence of NPC1L1 gene was amplified by 30 different PCR reactions. PCR products were barcoded and sequenced in a Junior-454 deep-sequencing platform (Roche). The resulting reads were aligned against the human genome (GRCh37) using BLAT algorithm and the variants were identified using GATK Unified genotyper. The functional consequence of the sequence variants were determined using SNPEff algorithm and association studies with patient phenotypes were performed using Plink suite. Results: 24 different sequence variants in NPC1L1 gene were identified in total in the 38 patients sequenced. 15 were small insertions and deletions (indels), whereas 9 of them constitute single nucleotide variants (SNV), from which 6 had been already reported in dbSNP database. According to a negative selection of deleterious sequence variants in the normal population, most of the identified variants were predicted to play synonymous or regulatory roles in the gene. Nevertheless, even with this limited sample size, association studies shown significative association between two of the sequence variants (a single nucleotide substitution and a single base deletion) with therapy response (rs186726309 and a novel SNV, Chr7: 44575955Del(G)). Additionally, a new SNV has been found which is not present in dbSNP database (Crh7: 44561370) and is present in a low frequency in our cohort; and produces a significant aminoacid change (S919C) in the coding region of the gene. Conclusions. Next-generation sequencing technologies to characterize variants in NPC1L1 gene identified new sequence variants that showed significant association with treatment response present in a population of CHC patients with and without antiviral-response a new variant with potential functional effect in NPC1L1 protein. Further research is needed to evaluate the potential prognostic value and therapeutics implications of these sequence variants.

Disclosures:

Manuel Romero-Gomez - Advisory Committees or Review Panels: Roche Farma,SA., MSD, S.A., Janssen, S.A., Abbott, S.A.; Grant/Research Support: Ferrer, S.A.

Javier Crespo - Board Membership: MSD, Roche, Janssen, Gilead

The following people have nothing to disclose: Joaquin Cabezas, Emilio Fábrega, Ignacio Varela, Jose Luis Fernandez Luna, Ana Fontalbo, Juan Antonio Gomez Gerique, Jose A. Del Campo, Angela Rojas, Angela Puente, Maria Teresa Arias, Marta García-Valdecasas

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Resistance Analysis of Genotype-1 and -3 HCV-Infected Patients Receiving MK-8742, a HCV NS5A Inhibitor with Potent Antiviral Activity

Rong Liu, Stephanie Curry, Patricia McMonagle, Robert B. Nachbar, Irene Pak, Patricia Jumes, Steve Ludmerer, Ernest Asante-Appiah, Daria Hazuda, Wendy W. Yeh, Anita Y. Howe;
Merck, Kenilworth, NJ

Background: MK-8742 is a small molecule inhibitor of Hepatitis C Virus (HCV) non-structural protein 5A (NS5A) that is being developed for the treatment of HCV infection. In vitro, MK-8742 has broad HCV genotypic activities and is potent against viral variants that are resistant to other 1st generation NS5A inhibitors. A Phase 1b, randomized, placebo-controlled study was conducted to assess the safety, pharmacokinetics and antiviral activity of MK-8742 in patients with chronic genotype (GT) -1 or -3 HCV infection. Methods: 48 adult males, with HCV RNA > 105 IU/mL and GT-1 or -3 chronic HCV infection without clinical evidence of cirrhosis, were randomized to receive placebo or MK-8742 from 5 to 50 mg (GT-1) or 10 to 100 mg (GT-3) once daily for 5 days. Plasma samples from baseline, the end of treatment and follow-up visits were collected and the full-length NS5A gene was analyzed by population sequencing. Selected samples were further analyzed with clonal sequencing to evaluate the distribution and linkage of resistance associated variants (RAVs). Results: MK-8742 has rapid inhibition leading to mean maximum viral load reductions of 3.7 - 5.1 log 10 IU/mL HCV RNA in GT-1 patients who received 5 - 50 mg daily doses. The durability of viral load decline following therapy was more sustained in GT-1 b patients than in GT-1 a patients at the same dose. Resistance associated variants, Y93H and M28V, were detected in two GT-1 patients prior to treatment. Despite the presence of baseline RAVs, these patients achieved >3 log viral load reduction. No viral breakthrough was observed during treatment. Post-baseline RAVs, M28A/T/V, Q30H/R, L31 F/V/I/M and Y93C/H/N/R, were detected at the end of treatment and at follow-up visits in GT-1 patients. Compared to GT-1, the antiviral response in GT-3 was less robust with a mean maximum viral load reduction of ∼3 log at 50 and 100 mg doses. Two GT-3 patients who had baseline RAVs (A30K/E/K/T) and received a sub-optimal 10 mg dose had a minimal viral load reduction. Post baseline RAVs, including A30E/K/T, L31I/F and Y93C/H/R. were detected at the end of treatment and at time points after treatment. Conclusions: MK-8742 exhibits potent antiviral activity during 5-day monotherapy in patients with chronic GT-1 and GT-3 HCV infection. Robust antiviral response was observed in the presence of baseline RAVs in 2 GT1 patients. RAVs at resistance loci common to other NS5A inhibitors were selected under antiviral pressure. The antiviral data support the continued clinical investigation of MK-8742 as a once-daily component of an all-oral, interferon-free regimen for the treatment of chronic HCV-infec-tion.

Disclosures:

Stephanie Curry - Employment: Merck

Patricia McMonagle - Employment: Merck and Co.

Robert B. Nachbar - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

Patricia Jumes - Employment: Merck; Stock Shareholder: Merck Steve Ludmerer - Employment: Merch & Co

Ernest Asante-Appiah - Employment: Merck

Daria Hazuda - Employment: Merck & Co.; Stock Shareholder: Merck & Co

Wendy W. Yeh - Employment: Merck & Co.

Anita Y. Howe - Employment: Merck Research Laboratory

The following people have nothing to disclose: Rong Liu, Irene Pak

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Pan-genotypic anti-HCV activity of SB 9200 assessed in the ‘capture-fusion’ replication assay

Morven E. Cunningham1, Joseph D. Wright1, Rajendra K. Pandey2, Anjaneyulu Sheri2, Seetharamaiyer Padmanabhan2, Rad-hakrishnan P. Iyer2, Graham R. Foster1;
1Queen Mary, University of London, London, United Kingdom; 2Spring Bank Pharmaceuticals, Milford, MA

SB 9200 is a novel, first-in-class anti-HCV drug which acts by enhancing the function of host cytosolic sensor proteins RIG-I and NOD2 that detect RNA viruses. It has shown potent activity against HCV G1a and G1 b replicons in vitro and is synergistic with other anti-HCV drugs such as telaprevir (NS3 protease inhibitor), NM283 (NS5B inhibitor), interferon and ribavirin. To explore the genotypic range of SB 9200, sensitivity of patient-derived HCV of different genotypes was tested in the recently developed capture-fusion assay, a novel HCV replication model that is highly predictive of clinical outcome of potential antivirals. Prestimulated THP-1 cells were infected with serum from donors with chronic HCV G1, 2, 3, 4 or 6, then fused with Huh7.5 cells and treated once with varying concentrations of SB 9200. For comparison, fused cells infected with G1 and G3 sera were treated with telaprevir or alisporivir. Hybrid cells were cultured for 5 days, before quantification of HCV RNA by PCR. Dose-response curves were used to calculate IC50 values. Results are given as mean ± s.d. and p values were calculated using the Mann-Whitney U test. SB 9200 inhibited replication of G1, 2, 3, 4 and 6 HCV strains in the fused cells. G3 isolates were significantly more sensitive to SB 9200 than to telaprevir (SB 9200 IC500.04 ± 0.01 μM, telaprevir IC500.12 ± 0.03 μM, p = 0.016). In G1 isolates, anti-viral potency of SB 9200 was comparable to that of alisporivir in this replication model (SB 9200 IC500.16 ± 0.12 μM, alisporivir IC500.14 ± 0.03 μM; p = 1.0). A similar pattern was seen in G3 isolates (SB 9200 IC500.04 ± 0.01 μM, alisporivir IC500.04 ± 0.02 μM; p = 0.4) (Figure 1). In a small number of isolates of other genotypes tested to date, all were sensitive to SB 9200 with G2 and G6 showing similar SB 9200 sensitivity to G3, and G4 strains more closely approximating G1. These results demonstrate activity of SB 9200 against a diverse range of HCV genotypes in vitro. Of note, this compound shows potent activity against patient-derived G3 isolates. These results support the potential role of SB 9200 as a pan-genotypic host-targeting anti-HCV agent.

Figure 1. Sensitivity of patient-derived G1 or G3 HCV to SB 9200, alisporivir or telaprevir in the capture-fusion assay. X axes show concentration of each drug, y axes show degree of inhibition of replication. Values are mean ± s.e.m.

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Disclosures:

Radhakrishnan P. Iyer - Employment: Spring Bank Pharmaceuticals, Inc

Graham R. Foster - Advisory Committees or Review Panels: GlaxoSmithKline, Novartis, Boehringer Ingelheim, Tibotec, Chughai, Gilead, Janssen, Idenix, GlaxoSmithKline, Novartis, Roche, Tibotec, Chughai, Gilead, Merck, Janssen, Idenix, BMS; Board Membership: Boehringer Ingelheim; Grant/Research Support: Chughai, Roche, Chughai; Speaking and Teaching: Roche, Gilead, Tibotec, Merck, BMS, Boehringer Ingelheim, Gilead, Janssen

The following people have nothing to disclose: Morven E. Cunningham, Joseph D. Wright, Rajendra K. Pandey, Anjaneyulu Sheri, Seetharamaiyer Padmanabhan

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Alisporivir treatment does not cause or exacerbate pancreatitis when co-administered with interferon-alpha and/or ribavirin in the cerulein-induced pancreatitis rat model

Dominique Brees1, Andre Cordier1, Joerg Andreas Mahl1, Jin Yi3, Francois Pognan4, Pierre Moulin1, David Ledieu1, Yoav E. Timsit2, Nikolai V. Naoumov4, Neeta G. Shenoy2, Jonathan Moggs1, Salah-Dine Chibout1;
1Novartis Institutes for BioMedical Research, Basel, Switzerland; 2Novartis Institutes for BioMedical Research, Cambridge, MA; 3Novartis Pharmaceuticals Corporation, East Hanover, NJ; 4Novartis Pharma AG, Basel, Switzerland

Alisporivir (ALV) is a cyclophilin inhibitor, in development for treatment of hepatitis C. Acute pancreatitis cases were reported in the clinical program - 6/1728 (0.35%) patients treated with ALV plus PegIFN/ribavirin; 2/489 (0.41%) patients treated with PegIFN/ribavirin only, and none with ALV IFN-free regimens. Previous studies with mouse models showed that genetic or pharmacological (ALV) inhibition of cyclophilin D reduces pancreas damage both in bile acid and in cerulein-induced models of pancreatitis. The purpose of this study was to determine the effects of ALV, interferon-alpha, and ribavirin on pancreatitis; we tested the outcome with these compounds, alone and in combinations, in a rat model of cerulein-induced pancreatitis. ALV (30 mg/kg/day) and ribavirin (100 mg/kg/day) were administered either alone or in combination orally to Sprague Dawley rats for 7 consecutive days prior to receiving rodent interferon alpha (500,000 IU/kg sub-cutaneous once per hour for 5x) and cerulein (50 ug/kg intra peritonea l 2× per hour). Cyclosporine A was included as a control. Pancreas was examined microscopically, a panel of biomarkers measured 3 and 24 hour after the last injection of cerulein, and the hepatic gene expression profile was determined at 24 hours. Administration of cerulein caused acinar degeneration, and in some animals ductular degeneration/necrosis in the pancreas. Cyclosporine A at ≥10 mg/kg caused a dose-related increase in severity of cerulein-induced acinar degeneration. Neither ALV nor ribavirin nor IFNα alone or in combination exacerbated the pancreas damage. However, co-administration of IFNα/ribavirin/ caused an increased incidence and severity of cerulein-induced pancreatic duct degeneration/necrosis. In contrast, ALV treatment was not associated with exacerbated acinar degeneration nor duct degeneration/necrosis when administered with cerulein or in combination with ribavirin and/or IFNα. In conclusion, Alisporivir alone or in combination with ribavirin or IFNα did not cause or exacerbate pancreatitis in the rat model of pancreatitis.

Disclosures:

Dominique Brees - Employment: Novartis

Andre Cordier - Consulting: Novartis

Joerg Andreas Mahl - Employment: Novartis Pharma AG; Stock Shareholder: Novartis Pharma AG

Pierre Moulin - Employment: Novartis Institutes of Biomedical Reserach

Yoav E. Timsit - Employment: Novartis; Management Position: Novartis; Stock Shareholder: Novartis

Nikolai V. Naoumov- Employment: Novartis Pharma AG, Novartis Pharma AG

Jonathan Moggs - Employment: Novartis

The following people have nothing to disclose: Jin Yi, Francois Pognan, David Ledieu, Neeta G. Shenoy, Salah-Dine Chibout

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Preclinical Characteristics of ACH-3422: A Potent Uridine Nucleotide Prodrug for Inhibition of Hepatitis C Virus Polymerase

Wengang Yang, Jason Wiles, Dharaben Patel, Steven Podos, Joanne L. Fabrycki, Yongsen Zhao, Lingling Jia, Guangwei Yang, Jose O. Rivera, Michael Elliot, Christopher Marlor, Akihiro Hashimoto, Godwin Pais, Venkat Gadachanda, Xiangzhu Wang, Dawei Chen, Qiuping Wang, Milind Deshpande, Mingjun Huang, Kathe Stauber, Avinash Phadke;
Achillion Pharmaceuticals, New Haven, CT

Background: While hepatitis C virus (HCV) NS5B RNA polymerase (NS5B Pol) nucleotide inhibitors impose a high genetic barrier to viral resistance, their activity as a mono-therapy is not sufficient to cure chronic hepatitis C (CHC). Discovery of ACH-3422, a novel HCV NS5B Pol uridine nucleotide inhibitor prodrug, for combination treatment with sovaprevir (PI) and ACH-3102 (NS5A inhibitor) is expected to produce an effective interferon-free therapy for CHC regardless of viral genotypes and patient characteristics. Here, we report the preclinical profile of ACH-3422. Methods: ACH-3422 potency was evaluated in cell lines harboring replicons with NS5B from different genotypes. Inhibition of NS5B Pol was assessed using the corresponding nucleoside triphosphate. Selectivity over human and other viral polymerases was examined with ACH-3422 in cell-based assays or its nucleoside triphosphate in cell free assays. Combination studies with sovaprevir and ACH-3102 were performed in replicon cell lines. Metabolism and PK properties were assessed by standard procedures. Safety was assessed in rats after oral administration for up to 14-days. Results: ACH-3422 displayed an EC50 of 50 nM in a cell line harboring genotype-1b replicon (compared to 150 nM for sofosbuvir) with a selective index of > 500. High potency was retained against a panel of replicons carrying NS5B from various genotypes. Biochemical assays with HCV NS5B Pol confirmed its nucleoside triphosphate acts as a potent non-obligate chain terminator. No antiviral activity against all viruses tested but BVDV was observed. No inhibition of human RNA and DNA polymerases was seen. Combinations in short-term with sovaprevir or ACH-3102 were not antagonistic and in long-term blocked the emergence of resistant variants at much lower concentrations compared to individual drugs. ACH-3422 was metabolized to its nucleoside triphosphate in animal and human hepatocytes and also in replicon cell lines. Oral dosing of ACH-3422 to animals resulted in high concentrations of its nucleoside triphosphate in liver and was well tolerated for 14-days at dose levels up to 250 mg/kg. Conclusions: ACH-3422 is a novel NS5B Pol uridine nucleotide inhibitor prodrug. In vitro, ACH-3422 or its nucleoside triphosphate demonstrates potent activity across different HCV genotypes and high selectivity for HCV NS5B Pol. In preclinical animal species, high liver concentrations of the nucleoside triphosphate were detected after oral dosing. With its profound effect to prevent the emergence of resistant variants in vitro, a clinical evaluation of ACH-3422 in combination with sovaprevir and ACH-3102 in hepatitis C patients is warranted.

Disclosures:

Wengang Yang - Employment: Achillion Pharmaceuticals; Stock Shareholder: Achillion Pharmaceuticals

Jason Wiles - Employment: Achillion Pharmaceuticals Michael Elliot - Employment: Achillion Pharmaceuticals

Xiangzhu Wang - Employment: Achillion Pharmceuticals Inc., Achillion Pharmceuticals Inc., Achillion Pharmceuticals Inc., Achillion Pharmceuticals Inc.

Dawei Chen - Stock Shareholder: Achillion

Milind Deshpande - Employment: Achillion Pharmaceuticals, Achillion Pharmaceuticals

Mingjun Huang - Employment: Achillion Pharmaceuticals

Kathe Stauber - Employment: Achillion Pharmaceuticals, Inc., Achillion Pharmaceuticals, Inc.

The following people have nothing to disclose: Dharaben Patel, Steven Podos, Joanne L. Fabrycki, Yongsen Zhao, Lingling Jia, Guangwei Yang, Jose O. Rivera, Christopher Marlor, Akihiro Hashimoto, Godwin Pais, Venkat Gadachanda, Qiuping Wang, Avinash Phadke

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Phosphoramidate Prodrugs of β-D-2″-C-Me-2,6-diaminopurine Ribonucleoside are Potent Inhibitors of Hepatitis C Virus (HCV) Replication Delivering Two Active Nucleoside Triphosphates

Raymond F. Schinazi1, Zhou Longhu1, Hongwang Zhang1, Maryam Ehteshami1, Sijia Tao1, Leda C. Bassit1, Justin Suesser-man1, Tony Whitaker2, Tami R. McBrayer2, Jong-Hyun Cho1, Sheida Amiralaei1, Jadd Shelton1, Mervi Detorio1, Steven Coats2;
1Emory University School of Medicine, and Veterans Affairs Medical Center, Atlanta, GA; 2RFS Pharma, Tucker, GA

BACKGROUND: We discovered novel β-D-2″-C-methyl-2,6-diaminopurine-ribonucleoside (DAPN) phosphoramidate prodrugs (PD) that inhibit HCV by generating two non-toxic bioactive nucleoside triphosphates (NTP) intracellularly. The major metabolite, DAPN-TP, was found to behave as an A-like analog. METHODS: DAPN-PD were compared to a known clinically toxic purine nucleoside INX-1 89 and non-toxic GS-7977. The median inhibitory concentration (IC50) and catalytic efficiency values for DAPN-TP and 2″-C-methyl-G-TP were evaluated against HCV NS5B polymerase. RESULTS: The DAPN-PD were pan-genotypic, effective against various HCV resistant mutants and HCV resistant variants could not be selected. DAPN-TP was the major metabolite in primary human hepatocytes, which has not been associated with cardiotoxicity versus 2″-C-Me-GTP and was 7-fold higher than found in Huh7 cells. Further analysis showed that unmasking of the DAPN-PD resulted in a non-toxic hydroxyphenyl carboxylic acid that is a known non-toxic metabolite of a common flavoring agent. DAPN-TP and 2″-C-Me-GTP were chain terminators for genotype 1 b HCV-pol with an IC50 of 3.6 and 0.46 μM, respectively, and had long intracellular half-lives. Single nucleotide incorporation assays revealed that DAPN-TP was incorporated opposite U, but not opposite C. INX-189 displayed cytotoxicity in various cells as well as bone marrow, elevated lactic acid and mitochondrial toxicity, which were not observed with various DAPN-PD when tested up to 50 μM. DAPN-PD showed prolonged stability in simulated gastric and intestinal fluids. A single oral dose of 50 mg/kg in 18 rats produced no adverse effects. CONCLUSIONS: DAPN-PD can deliver mostly DAPN-TP and smaller amounts of 2″-C-Me G-TP intracellularly. A DAPN prodrug has been selected for clinical development because of its low toxicity profile and its ability to deliver two active metabolites, thus simplifying HCV treatment.

Disclosures:

Raymond F. Schinazi - Stock Shareholder: RFS Pharma

Tony Whitaker- Employment: RFS Pharma, LLC

Tami R. McBrayer - Employment: RFS Pharma

Steven Coats - Employment: RFS Pharma

The following people have nothing to disclose: Zhou Longhu, Hongwang Zhang, Maryam Ehteshami, Sijia Tao, Leda C. Bassit, Justin Suesserman, Jong-Hyun Cho, Sheida Amiralaei, Jadd Shelton, Mervi Detorio

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Pharmacokinetic Interaction Between the HCV Protease Inhibitor MK-5172 and Midazolam, Pitavastatin, and Atorvastatin in Healthy Volunteers

Luzelena Caro1, Jennifer E. Talaty1, Zifang Guo2, Christina Reit-mann3, Iain P. Fraser3, Raymond Evers4, Dennis Swearingen5, Wendy W. Yeh3, Joan R. Butterton3;
1Clinical Drug Metabolism and Pharmacokinetics, Merck & Co., Inc., Whitehouse Station, NJ; 2Early Clinical Development Statistics, Merck & Co., Inc., White-house Station, NJ; 3Clinical Research, Merck & Co., Inc., White-house Station, NJ; 4Transporter-In Vitro Technology, Merck & Co., Inc., Whitehouse Station, NJ; 5Celerion, Tempe, AZ

Background: MK-5172, a potent, once-daily inhibitor of the hepatitis C virus (HCV) NS3/4A protease, is being developed for the treatment of chronic HCV infection. The aim of the study was to assess potential pharmacokinetic (PK) interactions of MK-5172 with midazolam (MDZ), pitavastatin, or atorvastatin in healthy subjects to determine the clinical relevance of MK-5172 as a CYP3A4 and organic anion-transporting polypeptide (OATP) inhibitor, and to evaluate the safety and tolerability of MK-5172 during co-administration. In vitro, MK-5172 is an inhibitor of OATP, breast cancer resistance protein (BCRP), and CYP3A4. MDZ and pitavastatin were used as probe substrates to evaluate potential interactions with CYP3A4 and OATP, respectively. Atorvastatin, a CYP3A4, OATP1B1, and BCRP substrate, is a widely-prescribed HMG-CoA reductase inhibitor that may be coadministered with MK-5172. Methods: This was an open-label, 3-part study in 29 healthy male and female subjects, ages 18-55 years. Part 1:11 subjects received a single dose of 2 mg/mL MDZ on Day 1, followed by 200 mg MK-5172 once-daily (QD) for 8 days, with a single dose of 2 mg/mL MDZ co-administered on Day 8. Part 2:9 subjects received a single dose of 20 mg atorvastatin followed by a 4-day washout. The subjects then received 200 mg MK-5172 QD for 8 days, followed by a single dose of 20 mg atorvastatin coadministered on Day 5. Part 3:9 subjects received a single dose of 1 mg pitavastatin followed by a 2-day washout. They then received 200 mg MK-5172 QD for 9 days, with a single dose of 1 mg pitavastatin coadministered on Day 7. Results: Coadministration of MK-5172 with MDZ, atorvastatin, or pitavastatin was safe and well-tolerated. MK-5172 increased the midazolam (MDZ) AUCO-oo with a geometric mean ratio (GMR, MDZ+MK-5172/MDZ) [90% confidence interval (CI)] of 1.34 [1.29, 1.39]. MK-5172 did not significantly impact the pitavastatin AUCO-oo, with a GMR (Pitava+MK-5172/Pitava) [90% CI] of 1.11 [0.91, 1.34]. MK-5172 increased the atorvastatin AUCO-oo and Cmax, with a GMR (Atorva+MK-5172/Atorva) [90% CI] of 3.00 [2.42, 3.72] and 5.66 [3.99, 9.45], respectively. Conclusions: MK-5172 did not significantly change pitavastatin PK, confirming that MK-5172 is not an OATP inhibitor in vivo. There was an approximate 30% increase in MDZ AUC when co-administered with MK-5172, suggesting that MK-5172 is a weak CYP3A4 inhibitor. There was an approximate 3-fold increase in atorvastatin AUC when co-administered with MK-5172, due to CYP3A4 inhibition and potentially BCRP inhibition. MK-5172 PK was not significantly impacted by co-administration with pitavastatin or atorvastatin.

Disclosures:

Luzelena Caro - Employment: Merck & Co., Inc.

Jennifer E. Talaty- Employment: Merck, Sharp, & Dohme

Zifang Guo - Employment: Merck & Co., Inc.

Christina Reitmann - Employment: Merck, Sharp & Dohme, Corp

Iain P. Fraser - Employment: Merck & Co.; Stock Shareholder: Merck & Co.

Raymond Evers - Employment: Merck

Wendy W. Yeh - Employment: Merck & Co.

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

The following people have nothing to disclose: Dennis Swearingen

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No Pharmacokinetic Interaction Between HCV Protease Inhibitor MK-5172 and HCV NS5A Inhibitor MK-8742 in Healthy Volunteers

Wendy W. Yeh1, Luzelena Caro1, Xiaobi Huang1, Eric Mangin1, Patricia Jumes1, Scott Rasmussen2, Joan R. Butterton1;
1Merck Sharp & Dohme Corp., Whitehouse Station, NJ; 2Celerion, Lincoln, NE

Background: MK-5172, a once-daily competitive inhibitor of the hepatitis C virus (HCV) NS3/4A protease with improved potency compared with the approved first generation protease inhibitors, and MK-8742, a HCV NS5A replication complex inhibitor with improved potency compared to first generation NS5A inhibitors, are being developed for the treatment of chronic HCV infection. Since these agents may be coadministered as a combination regimen for HCV, the present study evaluated the pharmacokinetic interactions and tolerability of MK-5172 and MK-8742 co-administration in healthy subjects. Methods: This was an open-label, multiple-dose study in 10 healthy adult male and female volunteers, ages 19-55 years. Since MK-5172 in HCV-infected patients demonstrates ∼2-fold higher exposure compared to healthy subjects, a 200 mg dose of MK-5172 in healthy subjects was used in this study to match the exposure of 100 mg dose (the intended Phase 3 dose) in HCV-infected patients. In Period 1, subjects received oral doses of 200 mg MK-5172 once daily on Days 1 to 7. Following a 7 day washout, subjects received oral doses of 20 mg MK-8742 once daily on Days 1 to 7 in Period 2. In Period 3, subjects were co-administered once daily oral doses of 200 mg MK-5172 and 20 mg MK-8742 on Days 1 to 8. Plasma PK samples were collected for the pharmacokinetic assessment of MK-5172 and MK-8742. Safety assessments included ECGs, vital signs, clinical laboratory tests, physical examination, and adverse event monitoring. Results: Co-administration of MK-5172 with MK-8742 was generally well-tolerated. Multiple oral doses of MK-5172 did not meaningfully change the steady-state AUC0-24h, Cmax, or C24h of MK-8742 with geometric mean ratios (GMRs) [90% confidence intervals (CIs)] for MK-8742 (MK-8742+MK-5172/MK-8742) of 1.01 [0.83, 1.24], 0.93 [0.76, 1.13], and 1.02 [0.83, 1.24], respectively. Multiple oral doses of MK-8742 did not meaningfully change AUC0-24h, Cmax, or C24h of MK-5172 (MK-5172+MK-8742/MK-5172) with GMRs [90% CIs] of 0.90 [0.63, 1.28], 0.87 [0.50, 1.52], and 0.94 [0.77, 1.15], respectively. Conclusions: Co-administration of MK-5172 and MK-8742 in healthy volunteers did not result in clinically significant drug-drug interactions. MK-5172 and MK-8742 were safe and well-tolerated when coadministered. Model-based predictions suggest that the DDI perpetrator and victim potentials of 20 and 50 mg (the intended clinical dose) doses of MK-8742 are expected to be comparable. These results suggest that no dose adjustments of MK-5172 or MK-8742 are needed in interferon-free, combination regimens containing these once-daily, direct acting antivirals in HCV-infected patients.

Disclosures:

Wendy W. Yeh - Employment: Merck & Co.

Luzelena Caro - Employment: Merck & Co., Inc.

Eric Mangin - Employment: Merck & Co., Inc.

Patricia Jumes - Employment: Merck; Stock Shareholder: Merck

Scott Rasmussen - Employment: Celerion, Inc

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

The following people have nothing to disclose: Xiaobi Huang

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MK-8742, a HCV NS5A Inhibitor with a Broad Spectrum of HCV Genotypic Activity, Demonstrates Potent Antiviral Activity in Genotype-1 and -3 HCV-Infected Patients

Wendy W. Yeh1, Concetta Lipardi1, Patricia Jumes1, Inge M. De Lepeleire1, Nick Van Den Bulk1, Luzelena Caro1, Xiaobi Huang1, Eric Mangin1, Robert B. Nachbar1, Edward J. Gane2, Serghei Popa3, Nelea Ghicavii3, Frank D. Wagner4, Joan R. Butterton1;
1 Merck Sharp & Dohme Corp., Whitehouse Station, NJ; 2Auckland City Hospital, Auckland, New Zealand; 3Republican Clinical Hospital, Chisinau, Moldova, Republic of; 4Charite Research Organisation GmbH, Berlin, Germany

Background: MK-8742 is an inhibitor of Hepatitis C Virus (HCV) non-structural protein 5A (NS5A) that is being developed for the treatment of HCV infection. MK-8742 has broad, potent HCV genotypic activity in vitro against viral variants that are resistant to other NS5A inhibitors in development. A Phase 1 b, randomized, placebo-controlled study was conducted to assess the safety, pharmacokinetics and antiviral activity of MK-8742 administered as 5 days of monotherapy in patients with genotype (GT) -1 or-3 HCV infection. Methods: 48 adult males, with HCV RNA > 105 IU/mL and GT-1 or -3 HCV infection without clinical evidence of cirrhosis, were randomized to receive placebo or MK-8742 from 5 to 50 mg (GT-1) or 10 to 100 mg (GT-3) once daily for 5 days (MK-8742: placebo ratio of 5:1 /panel). Safety and tolerability were evaluated using laboratory values, ECGs, and evaluation of adverse experiences (AEs). Antiviral efficacy was assessed using the Roche Cobas TaqMAN 2.0 plasma HCV RNA assay (lower limit of quantita-tion = 25 IU/mL). Results: Plasma HCV RNA declined rapidly after dosing with mean maximum reductions from baseline of 5.1 and 3.4 log10 IU/mL for GT-1 and GT-3 patients, respectively. The initial mean viral load (VL) reductions in GT-1a and 1b patients were similar among the 5- to 50-mg dosing groups, achieving >3 log VL decline after the first dose. No viral rebound occurred during dosing. A greater proportion of GT-1b patients achieved VL suppression below the limit of quanti-tation compared to GT-1a patients. The durability of VL decline was more sustained after cessation of dosing in GT-1 b patients than in GT-1 a patients at the same dose. Mean VL reductions after 5 days of MK-8742 in GT-3 patients were similar in 50-100 mg dose groups with more sustained virologic suppression after cessation of dosing in the 100 mg group. MK-8742 has a median Tmax of 2-4 hours on Day 5 with a mean apparent terminal t1 /2 of ∼19-27 hours. Steady-state was achieved within 5 days of MK-87425-100 mg QD administration. The range of accumulation ratios (day 5/1) for AUC0-24hr was 1.2-3.0. MK-8742 was generally well-tolerated, with all AEs transient and mild in intensity. The most common AE was headache. There were no clinically significant laboratory abnormalities, changes in vital signs or ECG readings. Conclusions: MK-8742 exhibits potent antiviral activity during 5 days of monotherapy in patients with GT-1 and GT-3 chronic HCV infection. The safety, pharmacokinetics, and antiviral data support the continued clinical investigation of MK-8742 as a once-daily component of an all-oral, interferon-free regimen for the treatment of chronic HCV-infection.

Disclosures:

Wendy W. Yeh - Employment: Merck & Co.

Patricia Jumes - Employment: Merck; Stock Shareholder: Merck

Inge M. De Lepeleire - Management Position: MSD (Europe) Inc. ; Stock Shareholder: Merck & co., Inc.

Luzelena Caro - Employment: Merck & Co., Inc. Eric Mangin - Employment: Merck & Co., Inc.

Robert B. Nachbar - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

Edward J. Gane-Advisory Committees or Review Panels: Roche, AbbVie, Novartis, Tibotec, Gilead Sciences, Janssen Cilag, Vertex, Achillion; Speaking and Teaching: Novartis, Gilead Sciences, Roche

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

The following people have nothing to disclose: Concetta Lipardi, Nick Van Den Bulk, Xiaobi Huang, Serghei Popa, Nelea Ghicavii, Frank D. Wagner

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Evaluation of second-generation NS5A inhibitors against hepatitis C virus by using NS5A replaced JFH-1 viruses and full-length infectious clones other than JFH-1

Asako Murayama, Nao Sugiyama, Takaji Wakita, Takanobu Kato;
Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan

About 3 % of world population is persistently infected with hepatitis C virus (HCV) and at increased risk of fatal chronic liver diseases such as cirrohsis and hepatocellular carcinoma. Because the efficacy of current therapy with pegylated IFN and ribavirin is insufficient and depends in part on viral genotypes, there is great interest in development of novel HCV-specific inhibitors. The development of new molecule that targets HCV protein called direct-acting antivirals is ongoing. Nonstructural protein 5A (NS5A) of HCV plays multiple and diverse roles in the viral lifecycle, and is currently recognized as a novel target for anti-viral therapy. In 2010, the first-generation NS5A inhibitor has been reported as to reduce the viral titer significantly after oral administration. However, it shows limited effects on genotype 2 HCV strains other than JFH-1 strain in cell culture system. Recently, to overcome this disadvantage, the second-generation NS5A inhibitors have been developed. The aim of this study was to evaluate the genotype-specific antiviral effects of these novel NS5A inhibitors. To assess the genotype-specificity of NS5A inhibitors, recombinant JFH-1 viruses replaced with NS5A of genotypes 1 (H77; 1a and Con1; 1b) and 2 (J6CF; 2a, MA; 2b and J8; 2b) were used. All these recombinant viruses were capable of replication and intracel-lular HCV core antigens of these viral RNA-transfected cells were at similar levels. Using these cell culture system, we examined the anti-viral effects of two derivatives of the second-generation NS5A inhibitor (ACH-3102). These compounds inhibited viral replications of wild-type JFH-1 and recombinant JFH-1 viruses with NS5A of genotype 1 (H77 and Con1), and the range of EC50 values is from 7.2 ± 1.2 pM to 38.4 ± 5.4 pM. They also inhibited the replications of recombinant JFH-1 viruses with NS5A of genotype 2 other than JFH-1 (J6CF, MA and J8), and the range of EC50 values is from 26.7 ± 1.7 pM to 198.0 ± 28.1 pM. We also confirmed the genotype-specific anti-viral effects of these NS5A inhibitors by using cell culture system with full-genome genotype 2 strains other than JFH-1 (J6cc and J8cc). The EC50 values of these compounds were lower or similar levels in J6cc and J8cc as compared with JFH-1. In conclusion, these second-generation NS5A inhibitors displayed improved potency against HCV genotype 2 strains compared with the first-generation NS5A inhibitor and will be expected to the NS5A inhibitors with pan-genotype activity.

Disclosures:

The following people have nothing to disclose: Asako Murayama, Nao Sugiyama, Takaji Wakita, Takanobu Kato

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Successful anti-SR-BI mAb therapy in humanized mice after challenge with HCV variants with partial in vitro resistance to SR-BI-targeting agents

Koen Vercauteren1, Naomi Van Den Eede1, Ahmed A. Mesalam1, Sandrine Belouzard2, Jean Dubuisson2, Thomas Pietschmann3, Geert Leroux-Roels1, Alfredo Nicosia4, Philip Meuleman1;
1Center for Vaccinology, Ghent University and Hospital, Ghent, Belgium; 2Center for Infection & Immunity of Lille, Lille, France; 3Department of Experimental Virology, Twincore, Hannover, Germany; 4CEINGE, Naples, Italy

Hepatitis C virus (HCV)-induced end-stage liver disease is currently the major indication for liver transplantation in the Western world. After transplantation the donor liver inevitably becomes infected by the circulating virus. We have previously shown that monoclonal antibodies (mAbs) against the HCV co-receptor scavenger receptor class B type I (SR-BI) inhibit HCV infection of different genotypes, both in cell culture and in humanized uPA-SCID mice. Anti-SR-BI mAb therapy was successful even when initiated several days after HCV exposure. These observations suggested that anti-SR-BI specific therapy may represent a novel therapeutic approach to prevent HCV reinfection of liver allografts. However, different HCV variants that seem to be less dependent on SR-BI have been described in the literature. Changes in the HCV glycoproteins such as deletion of E2 HVR1 (ΔHVR1), a single E2 substitution (G451 R) and single or multiple combined mutations in E1E2 (J6JFH1 Clone2 and mouse CD81-adapted virus) alter in vitro SR-BI usage. Compared to wild type virus, cell culture infectivity and propagation of these variants is much less efficiently blocked by anti-SR-BI therapy, which could negatively impact future therapeutic use. In this study we have evaluated whether humanized mice infected with HCV variants exhibiting decreased in vitro SR-BI-dependence remain responsive to anti-SR-BI mAb therapy. When given prior to viral challenge, anti-SR-BI mAbs prevented HCV infection in these mice and when administration was initiated several days after virus inoculation HCV RNA was cleared from the circulation. In addition, we tried to elucidate the mechanisms contributing to the discrepancies seen between in vitro and in vivo anti-SR-BI therapy efficacy. We did not find evidence supporting increased SR-BI-receptor usage of mouse-passaged viral particles. However, unlike wild type virus, the in vitro infectivity of virus variants with decreased SR-BI dependence was inhibited by both human HDL and VLDL. These lipoproteins considerably increased the antiviral potency of the mAbs against both the variants and the wild type virus. In conclusion, HCV variants that are less dependent on SR-BI for host cell entry and spread in vitro can be efficiently blocked by an anti-SR-BI mAb (designated mAb 16-71) in humanized uPA-SCID mice. This phenomenon might be explained by the presence of human lipoproteins in vivo that enhance the efficacy of the anti-SR-BI specific mAb. These properties, together with the fact that all these variants are more susceptible to neutralization by anti-HCV envelope antibodies, reduce their chance of emerging during anti-SR-BI therapy.

Disclosures:

Thomas Pietschmann - Advisory Committees or Review Panels: Janssen GmbH, Biotest AG; Speaking and Teaching: MSD Sharp & Dohme GmbH, Essex Pharma GmbH

The following people have nothing to disclose: Koen Vercauteren, Naomi Van Den Eede, Ahmed A. Mesalam, Sandrine Belouzard, Jean Dubuisson, Geert Leroux-Roels, Alfredo Nicosia, Philip Meuleman

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Effect of multiple oral doses of faldaprevir on the multiple dose pharmacokinetics of a combination oral tablet of ethinylestradiol and levonorgestrel in healthy pre-menopausal female volunteers

John P. Sabo1, Benjamin Lang2, Mabrouk Elgadi3, Fenglei Huang1;
1Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT; 2Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany; 3Boehringer Ingelheim Canada Ltd./Ltée., Burlington, ON, Canada

Aim: To evaluate the effect of multiple doses of 240 mg faldaprevir at steady-state on the pharmacokinetics (PK) of a combined oral contraceptive containing 30 μg ethinylestradiol (EE) and 150 μg levonorgestrel (LNG) in healthy premenopausal female volunteers. Methods: This open-label, 2-period, fixed sequence study started with a run-in period (between Days -56 and -28), where subjects received EE/LNG QD until Day 8. No treatment was provided on Days -7 to -1 to induce withdrawal bleeding. In Period 1, subjects received EE/LNG QD on Days 1 to 13 (Visit 3). Trough PK blood samples were taken on Days 1,11 and 12, with intensive PK blood sampling for EE and LNG on Day 13. Following completion of Period 1, subjects proceeded directly to Period 2 where they received EE/LNG QD and faldaprevir 240 mg QD on Days 14 to 21 (Visit 4; faldaprevir was dosed as 240 mg BID on Day 14 as a loading dose). Trough PK blood samples (faldaprevir, EE and LNG) were taken on Days 14, 19 and 20, with PK profiling blood samples obtained for EE and LNG on Day 21. Results: A total of 15/16 subjects completed the study and received all doses of study medication, with 1 subject prematurely discontinuing from study participation due to nausea. Overall, EE and LNG exposures were moderately higher when co-administered with faldaprevir than when administered alone (Table 1). Median t,/2 values were prolonged for both EE (2.4 hours longer) and LNG (4.7 hours longer) when co-administered with faldaprevir. Mean oral clearance and apparent volume of distribution of both EE and LNG were lower (∼30%) when co-administered with faldaprevir. Conclusions: Co-administration of faldaprevir and a combination oral contraceptive containing EE and LNG resulted in a moderate increase in exposure to both EE and LNG.

Table 6. 
  1. Test=EE/LNG and faldaprevir, Reference=EE/LNG, CI=confidence interval

PK parameterGeometric mean ratio Test/Reference [%]90% CI [%]
Ethinylestradiol (EE)  
AUCτ,SS141.0133.8-148.5
Cmax,SS114.8105.5-125.0
C24.SS171.4160.2-183.3
Levonorgestrel (LNG)  
AUCτ,SS140.5136.4-144.8
Cmax,SS115.3110.8-119.9
C24,SS153.9146.0-162.1

Disclosures:

John P. Sabo - Employment: Boehringer Ingelheim Pharmaceuticals, Inc.

Benjamin Lang - Employment: Boehringer Ingelheim Pharma GmbH & Co. KG

Mabrouk Elgadi - Employment: Boehringer Ingelheim

Fenglei Huang - Employment: Boehringer Ingelheim Pharmaceuticals, Inc

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Effect of steady-state faldaprevir on the pharmacokinetics of steady-state methadone and buprenorphine/naloxone in subjects on stable addiction management therapy

David Joseph1, Michael J. Schobelock1, Robert A. Riesenberg2, Bradley Vince3, Lynn R. Webster4, Abidemi Adeniji1, Mabrouk Elgadi5, Fenglei Huang1;
1Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT;2Atlanta Center for Medical Research, Atlanta, GA; 3Vince and Associates Clinical Research, Overland Park, KS; 4CRI Lifetree, Salt Lake City, UT; 5Boehringer Ingelheim Canada Ltd./Ltée., Burlington, ON, Canada

Aim: To evaluate the effect of faldaprevir at steady-state on the pharmacokinetics and pharmacodynamics of methadone or buprenorphine/naloxone in subjects on stable opioid maintenance therapy. Methods: This was an open-label study in subjects receiving a stable dose regimen of methadone (up to a maximum of 180 mg/day) or buprenorphine/naloxone (up to a maximum of 24 mg/6 mg per day). On Day 2, subjects received 480 mg faldaprevir (loading dose) followed by 240 mg QD faldaprevir on Days 3 to 9. Blood samples were taken on Days 1 and 9 for pharmacokinetic analysis for methadone and buprenorphine/naloxone (up to 24 h post-dose) and faldaprevir (up to 96 h post-dose). Pharmacodynamics of the opioid maintenance regimens were evaluated by the objective opioid withdrawal scale (OOWS) and subjective opioid withdrawal scale (SOWS). Results: Thirty four subjects entered and completed the study; 15 on methadone, 19 on buprenorphine/naloxone. Co-administration of faldaprevir with methadone or buprenorphine/naloxone resulted in geometric mean ratios for AUC0-24,ss″, Cmax,SS and C24,SS of si.2 for R-methadone and S-methadone, and ≤1.1 for buprenorphine and naloxone (Table 1). Similar faldaprevir exposures were observed in both the methadone and buprenorphine/naloxone treated subjects. There was no evidence of symptoms of withdrawal as evaluated by the validated OOWS or SOWS scores following co-administration of faldaprevir with methadone or buprenorphine/naloxone. Conclusions: No dose adjustment is required for methadone or buprenorphine/naloxone when co-administered with faldaprevir.

Table 7. 
  1. CI=confidence interval; gMean=geometric mean

  2. aAdjusted geometric mean ratio of (opioid drug with faldaprevir/opioid drug alone) × 100

PK parametergMean ratio [%]a90% CI [%]
R-Methadone  
AUC0-.24.ss118.1107.1-130.3
Cmax,SS114.1104.8-124.2
C24,SS111.899.2-125.9
S-Methadone  
AUC0-24,SS117.7106.9-129.5
Cmax,SS111.5100.4-123.7
C24,SS111.599.6-124.9
Buprenorphine  
AUC0-24,SS108.785.1-138.8
Cmax,SS92.473.5-116.3
C24,SS108.481.6-143.9
Naloxone  
AUC0-24,SS98.985.1-114.1
Cmax,SS94.378.0-114.1

Disclosures:

Michael J. Schobelock - Employment: Boehringer Ingelheim Pharmaceuticals Inc.

Lynn R. Webster - Advisory Committees or Review Panels: AstraZeneca, Boehringer Ingelheim, Covidien Mallinckrodt, Nektar Therapeutics, Orexo; Consulting: CVS Caremark, Jazz Pharmaceuticals, Neura Therapeutik, Quintiles, Ther-avance

Mabrouk Elgadi - Employment: Boehringer Ingelheim

Fenglei Huang - Employment: Boehringer Ingelheim Pharmaceuticals, Inc

The following people have nothing to disclose: David Joseph, Robert A. Riesen-berg, Bradley Vince, Abidemi Adeniji

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Inverse Modulation in Hepatic Expression of Interferon Receptor Complexes for Alpha and Lambda during HCV Infection are Associated with Altered Interferon Signaling Induction upon Treatment with Peginterferon Alfa-2a

Jacques Friborg1, Petra Ross-MacDonald2, Jian Cao2, Betsy J. Eggers1, Baiqing Lin1, Fiona McPhee1;
1 Virology, Bristol-Myers Squibb, Wallingford, CT; 2Transcriptional Profiling, Bristol-Myers Squibb, Pennington, CT

Background: Peginterferon Lambda-1a (Lambda), a Type III interferon (IFN), exerts potent antiviral activity through a unique receptor complex with limited cellular distribution outside the liver, and is expected to have a differentiated safety profile compared to peginterferon alfa (alfa). In a Phase 2b study of treatment-naive patients chronically infected with HCV representing genotypes (GT) 1 through 4, Lambda in combination with ribavirin (RBV) was associated with more rapid declines in HCV RNA compared to an alfa/RBV regimen. This correlated with improved viral response rates at Weeks 4 and 12 of treatment. To gain insight into the potential mechanisms of these early robust virologic responses with Lambda, we investigated the effects of HCV replication in vitro on the IFN signaling pathway in primary human hepatocytes (PHH). Methods: PHH obtained from healthy individuals were inoculated with cell culture adapted HCV (HCVcc) or with GT1 viruses derived from patient serum (HCVser). RNA was isolated from cells at multiple time-points and gene expression analysis performed using Affymetrix array profiling. Immunblotting analysis of HCV infected PHH was used to determine protein levels of IFN receptor subunits and to assess functionality of JAK-STAT signaling upon stimulation with alfa or Lambda. Results: Acute HCV infection induced a rapid down-regulation of the IFN alpha receptor subunit 1 (IFNAR1) transcript in PHH while transcriptional level of the unique IFN lambda receptor subunit IL28RA was increased. Immunoblotting analysis confirmed the repression of the IFNAR1 protein during infection with HCVcc or HCVser, which expression could be restored upon treatment with an NS3 protease inhibitor. Furthermore, induction of the IFN-responsive JAK-STAT signaling pathway was altered upon treatment with alfa whereas response to Lambda was not affected. Conclusions: Differential effects of HCV infection on expression of the IFN alpha and IFN lambda receptors in PHH may provide an explanation for the more robust early virologic response observed upon Lambda dosing in patients. The implications of this in vitro hepatic receptor modulation may be further explored during IFN-based combination therapy with direct-acting antivirals.

Disclosures:

Petra Ross-MacDonald - Employment: Bristol-Myers Squibb

Fiona McPhee - Employment: Bristol-Myers Squibb

The following people have nothing to disclose: Jacques Friborg, Jian Cao, Betsy J. Eggers, Baiqing Lin

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A Novel Uridine Nucleotide Prodrug, IDX20963, and Its Potential for Use in a Low-Dose Direct-Acting Antiviral Regimen for HCV

Lisa Lallos1, Xin-Ru Pan-Zhou1, Joseph McCarville1, Shouqi Luo1, Ilaria Serra1, Hassan Rashidzadeh1, Massimiliano LaColla1, Brenda Hernandez-Santiago1, John P. Bilello1, Christopher Chapron1, Francis Danehy1, Atyia N. Allen1, Michelle Camire1, Maria Seifer1, Kusum S. Gupta1, Jean-François Griffon2, Christophe C. Parsy2, Francois-Rene Alexandre2, Cyril B. Dous-son2, Nina L. Golitsina1, Sanjeev Bhadresa1, Ute Miner1, Roger Rush1, David N. Standring1;
1 Idenix Pharmaceuticals, Inc, Cambridge, MA; 2Idenix Pharmaceuticals, Inc., Montpellier, France

Purpose: This study characterized the activity, safety and early pharmacokinetic (PK) profile of IDX20963, a novel uridine liver-targeted nucleotide prodrug for the treatment of HCV. Methods: Anti-HCV activity was determined using recombinant HCV NS5B and in standard cell-based assays. Cytotoxicity was evaluated in a large panel of hepatic and non-hepatic mammalian cells and included galactose-cultured cells and TEM analysis of mitochondria. In vitro experiments were conducted using animal and human hepatocytes and subcellular fractions as well as human drug metabolizing enzymes and transporters. In vivo experiments were performed in mice, rats and monkeys following doses of 0.5 to 100 mg/kg. Results: The triphosphate (TP) of IDX20963 was active against HCV NS5B from genotypes 1 through 6 (97 to 250 nM), but not against cellular polymerases. IDX20963 was also active against HCV genotypes in cell-based assays, but inactive against 15 non-HCV viruses. IDX20963 was not cytotoxic to 19 mammalian cell types including cardiomyocytes or human bone marrow-derived progenitor cells with no evidence of mitochondrial toxicity. In resistance selection experiments NS5B S282T emerged slowly conferring mild resistance to IDX20963 (2.3 to 4.4 fold). IDX20963 was not cross-resistant to other direct-acting antiviral (DAA) classes and combination treatments of replicon cells with IDX20963 and DAAs from 4 other classes, including NS5A inhibitor samatasvir (IDX719), were overall additive. In vitro activity was not impacted by human serum proteins or the presence of common HBV or HIV drugs, and IDX20963 exhibited minimal inhibition of human drug metabolizing enzymes and transporters. Favorable PK properties included moderate to high bioavailability, rapid absorption, liver-targeted delivery with high, dose-proportional liver TP formation and a TP elimination half-life of approximately 24 hours in human hepatocytes. The dose-normalized TP level was ≥10 fold higher than its in vitro efficacy. No cardiac safety signals were observed in rat and monkey toxicology studies, including assessments of clinical pathology, histopathology, electrocardiograms, echocar-diograms and cardiac injury biomarkers. Conclusions: IDX20963 is a pan-genotypic HCV nucleotide prodrug with a favorable in vitro cytotoxicity profile, PK delivery, high liver extraction and activation to TP as well as minimal drug-drug interaction risk. These data suggest effective low-dose, once-daily use in combination with other DAA agents, such as samatasvir, and support advancing IDX20963 into phase I proof-of-concept clinical studies.

Disclosures:

Lisa Lallos - Employment: Idenix Pharmaceuticals, Idenix Pharmaceuticals, Idenix Pharmaceuticals, Idenix Pharmaceuticals

Xin-Ru Pan-Zhou - Employment: Idenix

Joseph McCarville - Employment: Indenix, Indenix, Indenix, Indenix; Stock Shareholder: Indenix, Indenix, Indenix, Indenix

Shouqi Luo - Employment: Idenix Pharmaceuticals, Inc. Ilaria Serra - Employment: Idenix Pharmaceuticals

John P. Bilello - Employment: Idenix Pharmaceuticals, Inc., Idenix Pharmaceuticals, Inc., Idenix Pharmaceuticals, Inc., Idenix Pharmaceuticals, Inc.

Christopher Chapron - Employment: Idenix Pharmaceuticals Atyia N. Allen - Employment: Idenix Pharmaceuticals Michelle Camire - Employment: Idenix Pharmaceuticals

Maria Seifer - Employment: Idenix Pharmaceuticals, Inc., Idenix Pharmaceuticals, Inc., Idenix Pharmaceuticals, Inc., Idenix Pharmaceuticals, Inc.

Kusum S. Gupta - Employment: Idenix Pharmaceuticals

Jean-François Griffon - Employment: Indenix; Stock Shareholder: Idenix

Christophe C. Parsy - Employment: Idenix Pharmaceuticals, Idenix Pharmaceuticals, Idenix Pharmaceuticals, Idenix Pharmaceuticals

Francois-Rene Alexandre - Employment: Idenix Cyril B. Dousson - Employment: Idenix

David N. Standring - Employment: Idenix Pharmaceuticals, Idenix Pharmaceuticals, Idenix Pharmaceuticals, Idenix Pharmaceuticals

The following people have nothing to disclose: Hassan Rashidzadeh, Massimiliano LaColla, Brenda Hernandez-Santiago, Francis Danehy, Nina L. Golitsina, Sanjeev Bhadresa, Ute Miner, Roger Rush

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Inhibition of hepatitis C virus in chimeric mice by sshRNAs: sequence analysis of surviving virus shows added selective pressure of combination therapy and provides strong evidence for an RNAi mechanism of action

Anne Dallas1, Heini Ilves1, Han Ma2, Daniel J. Chin2, Ian MacLach-lan3, Klaus Klumpp2, Brian H. Johnston1,4;
1SomaGenics, Inc., Santa Cruz, CA; 2Hoffmann-La Roche, Nutley, NJ; 3Tekmira Pharmaceuticals, Burnaby, BC, Canada; 4Pediatrics, Stanford University School of Medicine, Stanford, CA

We previously identified two short synthetic shRNAs (sshRNAs, SG273 and SG220) that target a conserved sequence within the internal ribosome entry site (IRES) of the hepatitis C virus (HCV), genotype (GT) 1. When formulated with lipid nanoparticles (LNP), these sshRNAs have been shown to inhibit HCV-linked gene expression and suppress viral replication in chimeric uPA-SCID mice infected with HCV by up to 2.5 log10. Viral load remained about 1 log10 below pre-treatment levels 21 days after the end of dosing. sshRNAs did not induce inflammatory cytokines, interferon, or ISG either in vitro or in vivo. Sequencing of HCV viral RNA amplified from serum after the 21-d follow-up period (500 nt surrounding the sshRNA target sites) showed that all mice treated with the active sshRNAs were altered in the respective target regions and virtually nowhere else in the region sequenced. In contrast, a control group that received an irrelevant (scrambled) sshRNA had no mutations in the region sequenced. SG220, the more potent of the active sshRNAs, selected mainly for mutations corresponding to its seed region, whereas the less potent SG273 selected for mutations in both its seed and non-seed regions. When mice were treated with a combination of both HCV sshRNAs, recovered viral sequences were found to be primarily mutated in the region of sequence overlap between the two sshRNAs, resulting in fewer mutations in the seed region of SG220. The ability of the most commonly selected mutations to confer resistance to the sshRNAs was confirmed in cell culture experiments by introducing those mutations into reporter plasmids in which the HCV IRES was linked to firefly luciferase expression. Strikingly, in a survey of 609 sequenced clinical isolates of HCV GT1 a and 1b in the European HCV database, the three nucleotide positions with the highest polymorphism in the 30-nt target region coincide with the three most frequent mutations induced by sshRNA treatment. These results demonstrate a direct antiviral activity, with fast and durable HCV suppression, and confirm action through a target-specific RNAi mechanism. They also suggest that 1 or 2 sshRNAs could be effective against HCV infection when combined with antiviral agents having different mecha-nism(s) of action, or when they are part of a cocktail comprising more than two sshRNAs.

Disclosures:

Anne Dallas - Employment: Somagenics; Patent Held/Filed: Somagenics; Stock Shareholder: Somagenics

Han Ma - Employment: Hoffmann-La Roche

Daniel J. Chin - Employment: Hoffmann-La Roche

Ian MacLachlan - Employment: Tekmira, Tekmira, Tekmira, Tekmira

Klaus Klumpp - Employment: Roche, Roche

Brian H. Johnston - Management Position: Somagenics, Inc.

The following people have nothing to disclose: Heini Ilves

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Pharmacokinetic Interactions Between the HCV Protease Inhibitor MK-5172 and Ritonavir-Boosted HIV Protease Inhibitors (Atazanavir, Lopinavir, Darunavir) in Healthy nVolunteers

Luzelena Caro1, Jennifer E. Talaty1, Zifang Guo2, Iain P. Fraser3, Henry U. Davis3, Terry O'Reilly4, Wendy W. Yeh3, Joan R. Butter-ton3;
1Clinical Drug Metabolism and Pharmacokinetics, Merck & Co., Inc., Whitehouse Station, NJ; 2Early Clinical Development Statistics, Merck & Co., Inc., Whitehouse Station, NJ; 3Clinical Research, Merck & Co., Inc., Whitehouse Station, NJ; 4Celerion, Lincoln, NE

Background: MK-5172, a potent, once-daily inhibitor of the hepatitis C virus (HCV) NS3/4A protease, is being developed for the treatment of chronic HCV infection in mono- and HCV/human immunodeficiency virus (HIV)-coinfected patients. The aim of this study was to assess the potential two-way pharmacokinetic (PK) interaction and tolerability of MK-5172 when coadministered with ritonavir (RTV)-boosted HIV protease inhibitors (PI), such as atazanavir/ritonavir (ATZr), lopinavir/ritonavir (LPVr), and darunavir/ritonavir (DRVr). MK-5172 is a substrate of breast cancer resistance protein (BCRP) and Pglycoprotein (P-gp) in vitro and an organic anion-transporting polypeptide (OATP) substrate, CYP3A4 substrate, and weak CYP3A4 inhibitor in vivo. ATZr, LPVr, and DRVr are substrates and potent inhibitors of CYP3A4/P-gp in vivo and potentially inhibitors of transporters (e.g., BCRP and OATP). Methods: This was an open-label, 3-parallel panel, 3-period study in 13 healthy male and female subjects per panel, ages 19-55 years. In Period 1, subjects received 200 mg of MK-5172 once-daily (QD) for 7 days, followed by a 7 day washout. In Period 2, subjects received either 300 mg ATZ/100 mg RTV QD, 400 LPV/100 mg RTV twice-daily (BID), or 600 mg DRV/100 mg RTV BID for 14 days, immediately followed by Period 3. In Period 3, 200 mg MK-5172 was coadministered with ATZr, LPVr, or DRVr for 7 days. Results: Coadministration of MK-5172 with ATZr, LPVr, and DRVr was safe and well-tolerated. MK-5172 did not significantly impact the lopinavir or darunavir PK, with a lopinavir AUC0-12 geometric mean ratio (GMR, LPVr+MK-5172/LPVr) [90% confidence interval (CI)] of 1.03 [0.92, 1.16] and darunavir AUC0-12 GMR (DRVr+MK-5172/DRVr) [90% CI] of 1.11 [0.99, 1.24]. MK-5172 modestly increased the atazanavir PK, with an AUC0-24 GMR (ATZr+MK-5172/ATZr) [90% CI] of 1.43 [1.30, 1.57]. MK-5172 exposures were significantly increased when coadministered with the ritonavir-boosted HIV PIs, with an AUC0-24 GMR (MK-5172+HIV PI-RTV/MK-5172) [90% CI] of 10.58 [7.78, 14.39] with ATZr, 12.86 [10.25, 16.13] with LPVr, and 7.50 [5.92, 9.51] with DRVr. Conclusions: Co-administration of MK-5172 with LPVr and DRVr did

not significantly affect lopinavir or darunavir exposures. Atazanavir exposure modestly increased when co-administered with MK-5172, which is consistent with weak CYP3A4 inhibition by MK-5172 in vivo. There was a significant increase in MK-5172 exposures when co-administered with LPVr, ATZr, and DRVr, which may be attributed to CYP3A4/P-gp inhibition and potentially inhibition of the transporter (e.g., OATP, BCRP)-medi-ated disposition of MK-5172.

Disclosures:

Luzelena Caro - Employment: Merck & Co., Inc.

Jennifer E. Talaty - Employment: Merck, Sharp, & Dohme

Zifang Guo - Employment: Merck & Co., Inc.

Iain P. Fraser - Employment: Merck & Co.; Stock Shareholder: Merck & Co.

Wendy W. Yeh - Employment: Merck & Co.

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

The following people have nothing to disclose: Henry U. Davis, Terry O'Reilly

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Role of BAFF in Prediction of Rapid Virological Response to PEG-IFN α-2a plus Ribavirin in Patients with Chronic Hepatitis C

Jing Liu, Zhen Xu, Wen-Xiong Xu, Zhi-Shuo Mo, Xiang Zhu, Zhi-liang Gao;
Infectious Diseases, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China

Background: B activating factor (BAFF) is a tumor necrosis factor super-family cytokine that is newly discovered. It plays an important role in the process of maturity and differentiation of B cells. There are not available that BAFF expression and the predictive value on treatment outcome in patients with chronic hepatitis C (CHC). Methods: 175 CHC patients were enrolled with PEG-IFNα-2a (180 μg, qw) / RBV (800-1200mg/d) treatment for at least 12w. IL28B SNP rs12979860 and rs8099917 and HCV genotype were detected at baseline, while serum ALT, AST, TBIL, HCV RNA loads and BAFF levels were detected before and 4w, 12w after treatment. BAFF levels were assessed on 58 health blood donors (HBD). To compare BAFF levels among these three groups, and analyze predictors to RVR in patients after antivirus treatments. Results: 1.BAFF levels in patients with RVR (median 2400.0pg/ml) were higher than that in patients without (median 1731.6pg/ml) and than that in HBD (median 1095.9pg/ml), successively, P<0.05. The lowest BAFF level was 724pg/ml in this study. 2. HCV genotype, IL28BSNP rs12979860 and rs8099917 genotype, HCV RNA and BAFF baseline were included in the logistic regression analysis and ROC was drawn to evaluate predict value of BAFF to RVR. After calculation IL28B SNP rs12979860 genotype and BAFF become influence factors to RVR. This was a logistic regression equation: Y=2.427-0.001 xBAFF-2.013×lL28BSNP rs12979860 (coded CC Genotype as 1 and non-CC as 0, Y=0.3614 according to the ROC cut-off point) . If a patient with CC genotype or non-CC genotype whose BAFF baseline were above 52.5pg/ml or 2065.5pg/ml, the AUROC achieved 79.6% (Figure 1). Conclusions: 1. BAFF expression in patients with hepatitis C-infected is higher than HBD. 2. BAFF is a predictor to RVR in patients with IL28BSNP rs12979860 non-CC genotype after antivirus therapy. There is higher RVR rate in patients with CC genotype regardless of others indexes.

Thumbnail image of

Figure l: ROC curve in prediction on BAFF to 1^ R after logistic regression analysis-Note: AUROC curve values (95% CI) were 0.796(0.684-0.908), cut-off point is 0.361, sensitivity is 0.904, specificity is 0-696, negative predictive value is 0.762,.P=0.000»

Disclosures:

The following people have nothing to disclose: Jing Liu, Zhen Xu, Wen-Xiong Xu, Zhi-Shuo Mo, Xiang Zhu, Zhi-liang Gao

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Antimicrobial Peptide LL-37 Deteriorate Infectivity of Hepatitis C Virus

Takanobu Kato1, Nao Sugiyama1, Asako Murayama1, Takuya Mat-sumura2, Masaaki Shiina3, Shinichi Asabe4, Takaji Wakita1, Michio Imawari5;
1Department of Virology II, National Institute of Infectious Diseases, Tokyo, Japan; 2Division of Gastroenterology, Department of Medicine, Showa University School of Medicine, Tokyo, Japan; 3Department of Gastroenterology and Hepatology, Shin-Yurigaoka General Hospital, Kawasaki, Japan; 4Saitama Medical Center, Jichi Medical University, Saitama, Japan; 5Research Institute for Gastrointestinal and Liver Diseases, Shin-Yurigaoka General Hospital, Kawasaki, Japan

Although recent studies indicate that supplementation of vitamin D significantly improves a sustained viral response by IFN-based therapy to chronic hepatitis C, detailed mechanisms for the role of vitamin D are not fully elucidated. Previously, we demonstrated that the metabolite of vitamin D, 25-hydroxyvita-min D, has the direct anti-viral effect against hepatitis C virus (HCV) targeting infectious virus production. Since vitamin D is known to be multi-functional, we reasoned that other anti-viral functions of vitamin D, especially through immunomodulatory activity, should be considered. The production of cathelicidin, one of antimicrobial peptides, has been demonstrated to be a part of the vitamin D-dependent antimicrobial pathway in the innate immunity. Cathelicidin is known to kill or inhibit the growth of microbial pathogens including mycobacteria and viruses directly. In this study, by use of HCV JFH-1-based cell culture system, we aimed to clarify the anti-HCV effects of the human cathelicidin, LL-37, produced by monocytes or macrophages in vitamin D dependent manner. HuH-7 cells were treated with LL-37 at the concentration of 10 μg/mL for 1 h and infected cell-culture generated HCV (HCVcc) at a multiplicity of infection of 0.5. HCV infection and production were estimated by measuring the intra- and extra-cellular HCV core antigen (Ag). By treatment of LL-37, intra- and extra-cellular HCV core Ag were reduced to about 30% as compared with untreated control. The cell viability assessed by WST-8 assay was not affected by this treatment. To see the effects on HCV replication, JFH-1 subgenomic replicon RNA transfected cells were treated with LL-37 in various concentrations. However, the inhibition of subgenomic replicon replication by LL-37 treatment was not detected. Next, to clarify the effects on HCV infection, HCVcc was treated with LL-37 at multiple concentrations in 37 °C for 1 h and infected into naïve HuH-7 cells after purification.

We found that the infectivity titer was diminished dose-dependently. The iodixanol gradient analysis revealed that the peak fraction of infectivity titer was disappeared by LL-37 treatment. In conclusion, the vitamin D associated antimicrobial peptide LL-37 deteriorated the infectivity of HCV. In addition to the direct anti-HCV effect of 25-hydroxyvitamin D, this anti-HCV effect of LL-37 might contribute to the improved efficacy of IFN-based therapy by supplementation of vitamin D.

Disclosures:

Takuya Matsumura - Grant/Research Support: Chugai pharmaceutical co.,ltd

Michio Imawari - Advisory Committees or Review Panels: Shionogi Pharmaceutical Co.; Consulting: Ajinomoto; Speaking and Teaching: Tanabe Mitsubishi Pharmaceutical Co., Yansen Pharma, Dainippon Sumitomo Pharmaceutical Co., Taisho Toyama Pharmaceutical Co., Tohre, Meiji Seika Pharma, GSK, MSD, Dai-ichi Sankyo, Chugai Pharmaceutical Co., Takeda Pharmaceutical Co., Ehzai

The following people have nothing to disclose: Takanobu Kato, Nao Sugiyama, Asako Murayama, Masaaki Shiina, Shinichi Asabe, Takaji Wakita

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Safety and Pharmacokinetics of Single and Multiple Oral Doses of MK-8742, an HCV NS5A Inhibitor, in Healthy Subjects

Eric Mangin1, Wendy W. Yeh1, Luzelena Caro1, Concetta Lipardi1, Anna Mitselos1, Iain P. Fraser1, Patricia Jumes1, Xiaobi Huang1, Daniel Dreyer1, Lucas M. Van Bortel2, Joan R. Butterton1;
1Merck & Co., Inc., West Point, PA; 2Drug Research Unit Ghent, Ghent, Belgium

Background: MK-8742, a hepatitis C virus (HCV) nonstructural protein NS5A replication complex inhibitor with improved potency compared to first generation NS5A inhibitors, is being developed for the treatment of chronic HCV infection. This study evaluated the safety and pharmacokinetics (PK) of single and multiple rising oral doses of MK-8742 in 48 healthy, male subjects (18 - 50 years of age). Methods: This was a double-blind, randomized, placebo-controlled, 2-part study. In Part 1, subjects received single oral doses of 5-400 mg of MK-8742 in the fasted state and a single 50 mg dose following a high fat breakfast. In Part 2, subjects received single oral 10-200 mg doses of MK-8742 once daily for 10 days in the fasted state. Plasma samples for PK were collected throughout the study period in both parts of the study. Safety assessments in both parts included ECGs, vital signs, clinical laboratory tests, physical examination, and adverse event monitoring. Results: Single and multiple doses of MK-8742 were generally safe and well-tolerated. No subjects discontinued due to adverse experiences. There was one non-drug related serious adverse experience; one subject in Part I had a left knee injury which required surgery. All other adverse experiences were mild to moderate in intensity and transient in nature. The most commonly reported adverse experience was headache. There were no clinically significant abnormalities in routine blood and urine chemistry panels, CBC, ECG, and physical examinations. In Part 1, single 5-400 mg oral doses of MK-8742 were rapidly absorbed (median Tmax of 3.5-4.0 hours). MK-8742 concentrations appeared to decline after Tmax in a bi-phasic manner, with the second phase initiating at about 12 hours. The mean terminal half-life (t1/2) was ∼14.5-19.9 hours. The mean AUC0-24hr and Cmax ranged from 80.8-3670 nM*hr and 6.33-340 nM respectively. MK-8742 exposure was reduced in the fed state (AUCO-oo fed/fast GMR of 0.67). AUC0-24hr was approximately less than dose-proportional across the 5-200 mg dose range. In Part 2, following 10-200 mg oral doses of MK-8742 once daily for 10 days, the absorption of MK-8742 was similar to Part 1. The mean t1/2 on Day 10 was 18.8-20.6 hours. The AUC0-24hr geometric mean achieved on Day 10 after 200-mg QD doses was 3.54 μM*hr. Steady state appears to have been reached within 10 days of dosing and the accumulation ratio for AUC0-24hr was 1.09-2.05 across the dose range studied. Conclusions: Single and multiple doses of MK-8742 were well-tolerated, and demonstrated pharmacokinetics amenable to once-daily dosing. Further investigation of this compound is warranted.

Disclosures:

Eric Mangin - Employment: Merck & Co., Inc.

Wendy W. Yeh - Employment: Merck & Co.

Luzelena Caro - Employment: Merck & Co., Inc.

Iain P. Fraser - Employment: Merck & Co.; Stock Shareholder: Merck & Co.

Patricia Jumes - Employment: Merck; Stock Shareholder: Merck

Lucas M. Van Bortel - Advisory Committees or Review Panels: Novartis; Independent Contractor: Merck, Actogenix, Daiichi Sankyo, Menarini; Speaking and Teaching: Recordati, Daiichi Sankyo

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.

The following people have nothing to disclose: Concetta Lipardi, Anna Mitselos, Xiaobi Huang, Daniel Dreyer

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Mass balance, metabolic profile and the role of hepatic and bacterial enzymes in the metabolism of the HCV polymerase inhibitor, deleobuvir (BI 207127)

Rucha S. Sane1, Lin-Zhi Chen1, Elsy Philip1, Sean Regan1, Hlaing Maw1, Hongbin Yu1, Arti Mathur1, Yongmei Li1, Debra A. Man-darino2, John P. Sabo1;
1Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT; 2Covance Clinical Research Unit Inc, Madison, WI

Aim: Deleobuvir (BI 207127, DLV) is a specific, potent, reversible non-nucleoside inhibitor of HCV polymerase intended for interferon-free combination therapy with ribavirin and faldaprevir. This study was conducted to determine the pharmacokinetics of DLV and total radioactivity including mass balance, excretion pathways, metabolism and mechanisms for metabolite formation after oral administration of [14C]-DLV. Methods: Twelve healthy male volunteers received a single oral dose of 800 mg [14C]-DLV (100 μCi). Blood, plasma, urine, feces and saliva were collected for 6 to 10 days and analyzed for radioactivity. Metabolite profiling was performed using plasma, urine and fecal samples by radiochromatography, and metabolite identification was based on LC/MS/MS. Quantitation of DLV and its major plasma metabolites, was performed using synthetic standards. Exposure of the major metabolites in humans was compared to that in rats and mice. In vitro studies with recombinant CYP450 enzymes, UGT enzymes and gut bacteria were performed to identify enzymes responsible for the metabolism of DLV. Results: Mean recovery of radioactivity was high (93.4% over 216 h). Most of the radioactivity was recovered in feces (93.3%), with minimal excretion into urine (0.146%). DLV represented the major circulating species in plasma (∼63.0% of total AUC). There were two major circulating metabolites, BI 208333, an acyl glucuronide conjugate representing 20.2% of total AUC, and CD 6168, an alkene reduction metabolite, representing 15.3% of total AUC. DLV, BI 208333 and CD 6168 demonstrated similar time courses of plasma exposure, with median tmax values in the range of ∼3 to 6 h and geometric mean terminal half-life of ∼3 h. DLV and CD 6168 were also the predominant components in feces representing 30.4% and 34.6% of radioactivity dose, respectively. Other fecal metabolites included three hydroxylated metabolites, which are most likely secondary metabolites of CD 6168, each representing <10% of the radioactive dose. In vitro data indicated that UGT1 A1 was predominantly responsible for formation of BI 208333, whereas gut bacteria, but not microsomal or cytosolic hepatic enzymes, were responsible for the reduction of DLV to form CD 6168. Conclusions: DLV demonstrated good recovery and mass balance in healthy male volunteers. Two major circulating metabolites of DLV were identified, BI 208333 and CD 6168, which had similar pharmacokinetic profiles to that of the parent. Both metabolites were adequately represented in the nonclinical toxicity studies. Metabolism and excretion into feces are the major elimination routes for DLV.

Disclosures:

Lin-Zhi Chen - Employment: Boehringer Ingelheim Pharmaceuticals

Sean Regan - Employment: Boehringer Ingelheim Pharmaceuticals Inc.

Hongbin Yu - Employment: Boehringer Ingelheim Pharmaceuticals, Inc.

John P. Sabo - Employment: Boehringer Ingelheim Pharmaceuticals, Inc.

The following people have nothing to disclose: Rucha S. Sane, Elsy Philip, Hlaing Maw, Arti Mathur, Yongmei Li, Debra A. Mandarino

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Pharmacokinetic Interaction Between the HCV Protease Inhibitor MK-5172 and Efavirenz in Normal Healthy Volunteers

Jennifer E. Talaty1, Luzelena Caro1, Wendy W. Yeh2, Iain P. Fraser4, Zifang Guo3, Kristin Butterfield5, Christina Reitmann4, Katherine M. Dunnington6, Bruce J. DeGroot6, Stephen P. Young-berg6, Joan R. Butterton2;
1Clinical Drug Metabolism and Pharma-cokinetics, Merck, Sharp & Dohme, West Point, PA; 2Clinical Pharmacology, Merck, Sharp & Dohme, Boston, MA; 3Early Clinical Development Statistics, Merck, Sharp & Dohme, North Wales, PA; 4Clinical Pharmacology, Merck, Sharp, & Dohme, Rahway, NJ; 5Clinical Pharmacology, Merck, Sharp, & Dohme, North Wales, PA; 6Celerion, Lincoln, NE

Background: MK-5172, a reversible, noncovalent, competitive inhibitor of the hepatitis C virus (HCV) NS3/4A protease, is being developed for the treatment of chronic HCV infection. Efavirenz (EFV) is a non-nucleoside reverse transcriptase inhibitor specific for human immunodeficiency virus-1 (HIV). Chronic HCV infection is a major cause of morbidity and mortality in patients coinfected with HIV and HCV. The aim of the present study was to evaluate potential pharmacokinetic (PK) interactions as well as safety and tolerability of MK-5172 (a CYP3A4 substrate and weak CYP3A4 inhibitor) when co-administered with EFV (a CYP3A4 inducer and CYP3A substrate) in healthy subjects. Methods: This was an open-label, fixed-sequence, multiple-dose study in 12 healthy adult male and female volunteers, ages 19-55 years. Each subject received oral doses of 200 mg MK-5172 once daily for 7 days, followed by a 7 day washout. In Period 2, subjects received oral doses of 600 mg EFV once daily for 14 days. In Period 3, which commenced immediately after Period 2, subjects were coadministered oral doses of 200 mg MK-5172 and 600 mg EFV once daily for 7 days. Blood samples were obtained for EFV PK evaluation on Day 14 in Period 1 and Day 7 in Period 3 and for MK-5172 on Day 7 in Periods 1 and 2. Trough samples were also collected for both compounds to assess achievement of steady state. Safety assessments included electrocardiograms, vital signs, clinical laboratory tests, physical examination, and adverse event monitoring. Results: Co-administration of MK-5172 with EFV was safe and well-tolerated. Steady-state was achieved for both compounds alone and with coadministration. Multiple oral doses of EFV decreased the steady-state AUC0-24, Cmax, and C24h of MK-5172 with geometric mean ratios (GMRs, MK-5172+EFV/MK-5172) [90% confidence intervals (CIs)] of 0.16 [0.12, 0.28], 0.30 [0.25, 0.37], and 0.12 [0.08, 0.19], respectively. Multiple oral doses of MK-5172 did not meaningfully change the steady-state AUC0-24, Cmax, or C24h of EFV with GMRs (EFV+MK-5172/EFV) [90% CIs] of 1.00 [0.96, 1.05], 0.93 [0.88, 0.98], and 1.03 [0.99, 1.08], respectively. Conclusions: There was a decrease in MK-5172 PK in normal healthy volunteers when coadministered with EFV, likely due to CYP3A4 induction by EFV. There was no clinically meaningful effect of MK-5172 on the PK of EFV.

Disclosures:

Jennifer E. Talaty- Employment: Merck, Sharp, & Dohme

Luzelena Caro - Employment: Merck & Co., Inc.

Wendy W. Yeh - Employment: Merck & Co.

Iain P. Fraser - Employment: Merck & Co.; Stock Shareholder: Merck & Co.

Zifang Guo - Employment: Merck & Co., Inc.

Kristin Butterfield - Employment: Merck, Sharp & Dohme

Christina Reitmann - Employment: Merck, Sharp & Dohme, Corp

Stephen P. Youngberg - Employment: Celerion, Inc.

Joan R. Butterton - Employment: Merck & Co., Inc.

The following people have nothing to disclose: Katherine M. Dunnington, Bruce J. DeGroot

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No Clinically-Relevant Interactions Between Asunaprevir and Selective Serotonin Reuptake Inhibitors (Escitalo-pram and Sertraline) in Healthy Subjects

Tushar Garimella1, Robert Adamczyk1, Peter Hu1, Michele Stonier1, Hamza Kandoussi2, Elizabeth Colston1, Timothy Eley1;
1Research and Development, Bristol-Myers Squibb, Hopewell, NJ; 2Research and Development, Bristol-Myers Squibb, Lawrenceville, NJ

Background Chronic HCV infection and some HCV therapies are associated with psychiatric illness, including depression. Asunaprevir (ASV) is a selective HCV NS3 protease inhibitor with activity in vitro against HCV genotypes 1, 4, 5 and 6, demonstrated clinical efficacy and safety as part of multiple (including all oral) regimens, and a pharmacokinetic (PK) profile supportive of BID dosing without food restrictions (softgel capsule). ASV is a weak to moderate precipitant of drug-drug interactions (DDIs). As it is anticipated that ASV will be coadministered with antidepressants (e.g. escitalopram [ESC] and sertraline [SER]) it is important to understand the potential for DDIs. Methods Healthy fasted subjects (age 25-55 years; BMI 18-32 kg/m2) received oral ESC 10 mg QD (Cohort 1) or SER 50 mg QD (Cohort 2) on Days 1-7 and 25-31; all received oral ASV 100 mg BID on Days 15-31. Blood samples for PK analyses of ESC and SER (24h) were collected on Days 7 and 31; samples for ASV (12h) were collected on Days 24 and 31. Geometric mean ratios (GMR) and 90% confidence intervals (CI) for ESC, SER and ASV maximum plasma concentrations (CMAX) and area under the plasma concentration time curve in one dosing interval (AUCMAXTAU) were calculated. The limits of the no effect boundary of ASV on the exposure of ESC or SER (0.7-1.7) and ESC or SER on ASV (0.5-2.0) were set by review of literature and clinical data. Results Sixteen subjects (14 male) were enrolled in Cohort 1 and 15 completed the study. Eighteen subjects (15 male) were enrolled in Cohort 2 and all completed the study. Median age in Cohort 1 was 37 years (range 25-52) while that in Cohort 2 was 33 years (range 25-53). ASV did not affect the exposures of either ESC or SER, and ESC and SER did not affect the exposure of ASV to a clinically-relevant degree; 90% CIs of the GMRs for CMAX and AUCTAU were within the set no effect boundaries (see Table). There were no serious adverse events (AEs). One subject in Cohort 1 reported two moderate AEs (agitation [led to discontinuation] and insomnia) during combination treatment; all other

AEs were of mild intensity and resolved prior to study discharge. Conclusions Co-administration of ASV with ESC or SER is well tolerated and does not result in clinically-significant changes in ASV, ESC or SER exposure. ASV and ESC or SER can be co-administered without dose adjustments.

Table 8. 
 GMR (90% CI)
Treatment comparisonCmaxAUCTAU
Cohort 1 (n=l5) ESC + ASV vs ESC alone ESC + ASV vs ASV alone0.97(0.92, 1.02)0.87 (0.65, 1.18)0.95(0.91,0.98)0.92 (0.76, 1.12)
Cohort 2 (n=l8) SER + ASV vs SER alone SER + ASV vs SER alone* SER + ASV vs ASV alone0.81 (0.67, 0.97) 0.89 (0.82,0.96) 0.94 (0.70, 1.28)0.79 (0.67,0.94) 0.86 (0.79,0.94) 0.88 (0.70, 1.11)
*Excluding outlier (subject had ∼5-7 fold lower CMAX and AUC for in the SER+ASV arm compared with the SER alone arm. ASV exposure was not affected)

Disclosures:

Tushar Garimella - Employment: Bristol Myers-Squibb; Stock Shareholder: Abbvie

Robert Adamczyk-Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers Squibb

Michele Stonier - Employment: Bristol Myers Squibb Elizabeth Colston - Employment: Bristol-Myers Squibb

Timothy Eley-Employment: Bristol-Myers Squibb; Stock Shareholder: Bristol-Myers Squibb

The following people have nothing to disclose: Peter Hu, Hamza Kandoussi

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Phenotypic antivirogram of clinical strains of hepatitis C virus using cryopreserved primary human hepatocytes

Tony Durand1, Patrice Bruscella2, Claire Gondeau5, Mehdi Lahmar3, Michel Ventura4, Cyrille Feray2;
1IMAD, Hotel-Dieu, Nantes, France; 2INSERM 955 team 18, Hopital Henri Mondor, Creteil, France; 3Drug research, Vivalis, Saint Herblain, France; 4UMR 5234, Institut National des Sciences Biologiques, Bordeaux, France; 5Inserm U1040, Institute of Research in Biotherapy, Bordeaux, France

The development of anti-HCV drugs is based on HCV subgenomic replicons and on the JFH1 strains which both replicate in huh7 cell lines. The screening of antiviral drug on the replication of clinical seric strains (HCVser) remain elusive. Primary cultures of human hepatocytes (PHH) are known to permit a short-term, low and poorly reproducible replication of some HCVser. Interestingly, cryopreserved PHH have not been tested to establish a phenotypic antivirogram. Methods HCVser from genotype 1 to 4 harvested in 39 viremic patients. One of them developed telaprevir resistance (strain M). Telaprevira protease inhibitor, acyl pyrrolidine a polymerase inhibitor and 2 candidate anti-polymerase drugs were tested. All these molecules had an IC50 < 100 nmol/L in HCV replicon and JFH1 models. Fresh PHH (fPHH) and cryopreserved PHH (cPHH) were obtained after liver resection (Biopredic Inc, Rennes). All experiments were in triplicate or more. HCV RNA in cell culture and supernatant were measured by RT-PCR 3 days post inoculation. Results In fPHH, JFH1 lead to 3 to 4 log/μg at day 3. Using 7 different fPHH donors, only 3/39 HCVser lead to a low-level replication (log2-3) in more than one experiment. Using 4 different cPHH donors, 3 batches reproducibly permitted a log 4-5 viral load for 8/10 tested HCVser and for JFH1. TPV and pyrrolidine inhibited most HCVser (Genotype 1) with an IC50 which was 3 to 10-fold higher than those observed for replicon or JFH1. TPV resistance of the strain M was confirmed. The molecules A and B did not reduce the replication of any clinical strain in CPHH or in fPHH and lead to hepatocytes necrosis. No clear relationship was found with IL28B genotype of PHH. Finally, the cellular mortality was lower for cPHH than forfPHH. Conclusion In the era of direct antiviral molecules, cPHH authorize a significant replication of different HCVser and reliably permit the comparison of different antiviral compounds on most natural HCV strains. A striking finding was that some compound could have an antiviral activity in artificial HCVcc models but not in HCVser.

Disclosures:

Mehdi Lahmar - Employment: Vivalis/valneva

The following people have nothing to disclose: Tony Durand, Patrice Bruscella, Claire Gondeau, Michel Ventura, Cyrille Feray

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Pharmacokinetic Interaction Between the HCV Protease Inhibitor MK-5172 and IV and Oral Rifampin in Healthy Volunteers

Luzelena Caro1, Jennifer E. Talaty1, Zifang Guo2, Kristin Butter-field3, Thomayant Prueksaritanont4, Scott Rasmussen5, Iain P. Fraser3, Wendy W. Yeh3, Joan R. Butterton3;
1Clinical Drug Metabolism and Pharmacokinetics, Merck & Co., Inc., Whitehouse Station, NJ; 2Early Clinical Development Statistics, Merck & Co., Inc., Whitehouse Station, NJ; 3Clinical Research, Merck & Co., Inc., Whitehouse Station, NJ; 4Preclinical Drug Metabolism and Pharmacokinetics, Merck & Co., Inc., Whitehouse Station, NJ; 5Cele-rion, Lincoln, NE

Background: MK-5172, a reversible, non-covalent, competitive inhibitor of the hepatitis C virus (HCV) NS3/4A protease, is being developed for the treatment of chronic HCV infection. The aim of the study was to evaluate the potential pharmacokinetic (PK) interactions, as well as safety and tolerability of MK-5172, when co-administered with IV and oral rifampin (RIF) in healthy subjects. MK-5172 is a substrate of organic anion-transporting polypeptide (OATP) and breast cancer resistance protein (BCRP) in vitro and a CYP3A4/P-glycoprotein (P-gp) substrate in vivo. Rifampin is a potent inducer of CYP3A4/P-gp following multiple-dose administration, an inhibitor of P-gp with acute administration, and an inhibitor of OATP/BCRP. Therefore, the study also assessed the clinical significance of the OATP/BCRP and CYP3A4/P-gp pathways for MK-5172. Methods: This was an open-label, fixed-sequence, multiple-dose study in 12 healthy male and female volunteers, ages 19-55 years. In Period 1, subjects received a single intravenous (IV) dose of 600 mg RIF coadministered with a single oral dose of 200 mg MK-5172, followed by a 7-day washout. In Period 2, once-daily (QD) doses of 200 mg MK-5172 were administered for 8 days. Period 2 was immediately followed by Period 3, in which subjects received 200 mg MK-5172 QD coadministered with 600 mg QD oral doses of RIF for 14 days. Results: Coadminis-tration of MK-5172 with RIF was safe and well-tolerated. A single IV dose of RIF increased the MK-5172 AUC0-24, Cmax, and C24, with geometric mean ratios (GMRs, MK-5172+RIF/MK-5172) [90% confidence intervals (CIs)] of 12.61 [10.83, 14.67], 10.94 [8.92, 13.43], and 1.77 [1.40, 2.24], respectively. A single dose of oral RIF increased the MK-5172 steady-state AUC0-24, Cmax, and C24 with GMRs (MK-5172+RIF/MK-5172) [90% CIs] of 8.35 [7.38, 9.45], 6.52 [5.16, 8.24], and 1.62 [1.32, 1.98], respectively. Multiple oral doses of RIF did not statistically impact the MK-5172 steady-state AUC0-24 or Cmax with GMRs (MK-5172+RIF/MK-5172) [90% CI] of 0.93 [0.75, 1.17] and 1.16 [0.82, 1.65], respectively, but decreased the MK-5172 C24h with a GMR [90% CI] of 0.15 [0.11, 0.20]. Conclusions: There was a significant increase in MK-5172 PK when MK-5172 is coadministered with a single IV or oral dose of RIF, which may be primarily attributed to inhibition of OATP by RIF. There was no significant effect of oral 600 mg QD RIF on MK-5172 AUC and Cmax, but a significant decrease in C24h, likely due to a net-effect of OATP inhibition and CYP3A4/P-gp induction by multiple oral RIF doses. These results suggest that MK-5172 is an OATP substrate and confirm that MK-5172 is a CYP3A4/P-gp substrate.

Disclosures:

Luzelena Caro - Employment: Merck & Co., Inc.

Jennifer E. Talaty - Employment: Merck, Sharp, & Dohme

Zifang Guo - Employment: Merck & Co., Inc.

Kristin Butterfield - Employment: Merck, Sharp & Dohme

Thomayant Prueksaritanont - Employment: Merck Sharp & Dohme Corp

Scott Rasmussen - Employment: Celerion, Inc

Iain P. Fraser - Employment: Merck & Co.; Stock Shareholder: Merck & Co.

Wendy W. Yeh - Employment: Merck & Co.

Joan R. Butterton - Employment: Merck Sharp & Dohme Corp.; Stock Shareholder: Merck Sharp & Dohme Corp.