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STEP Trial Efficacy Analyses HVTN Full Group Meeting November 7, 2007 1 07-Nov-2007 Acknowledgements Special thanks to the team of biostatisticians who have worked relentlessly on preparing these analyses Merck • • • • Devan Mehrotra Robin Mogg David Li Jonathan Hartzel HVTN • Steve Self • Peter Gilbert 2 07-Nov-2007 Efficacy Hypothesis • Primary Hypotheses (Ad5 ≤200 Stratum) 1) Infection endpoint: Among subjects with baseline Ad5 titers ≤200, those who receive the MRKAd5 HIV-1 gag/pol/nef vaccine will subsequently have a lower likelihood of acquiring HIV-1 infection compared to those who receive placebo and/or 2) Viral load endpoint: Among subjects with baseline Ad5 titers ≤200 who subsequently become HIV-1 infected, those who receive the MRKAd5 HIV-1 gag/pol/nef vaccine will have a smaller average viral load set-point (HIV-1 RNA at ~3 months postdiagnosis) compared to those who receive placebo • Secondary hypotheses were same as primary hypotheses, but in the overall study population (Ad5 ≤200 and Ad5 >200 strata combined) 3 07-Nov-2007 Two analysis populations • Modified intent-to-treat (MITT) population includes all participants who: – Were HIV seronegative on day of randomization – Received at least one study injection – Best population for evaluating infection endpoint • Per protocol (PP) population includes all participants who: – Were HIV seronegative at Week 12 study visit – Received at least the first 2 study injections in window – Were not “protocol violators” • Met entry criteria for study • Did not receive non-study vaccine (eg, flu shot) in prohibited window • Had a confirmatory visit following diagnosis of HIV infection • Did not receive ART before confirmatory visit – Best population for simultaneously evaluating both endpoints 4 07-Nov-2007 Analysis strategy – Event driven • Newly acquired HIV infection • Viral load (VL) 3 months post-diagnosis – Planned interim analyses in addition to final analysis • 1st interim analysis triggered when 30 “per-protocol” events identified in Ad5 ≤ 200 stratum • Futility to be assessed at each interim analysis – p-value > 0.50 (1-tailed) for EACH endpoint » Trend favoring placebo for each endpoint » Likelihood of meeting either endpoint at final analysis is very low (conditional power <10%) 5 07-Nov-2007 Statistical Power • In Ad5 ≤ 200 study population – – With ~50 PP events, 80% power to detect 60% vaccine efficacy for acquisition endpoint With ~30 PP events, 80% power to detect ~1-log reduction in VL 3 months post-diagnosis (first interim) • In overall study population – – With ~100 PP events, 80% power to detect 50% vaccine efficacy for acquisition endpoint With ~60 PP events, 80% power to detect ~0.75-log reduction in VL 3 months post-diagnosis 6 07-Nov-2007 Case split for infection endpoint Primary analysis (Ad5 ≤ 200) Vaccine Total MITT cases Cases included in PP efficacy analysis 24 19 Placebo 21 11 Primary dataset reviewed by DSMB 7 07-Nov-2007 Case split for infection endpoint Primary analysis (Ad5 ≤ 200) Vaccine Total MITT cases Cases included in PP efficacy analysis Cases excluded from PP efficacy analysis 24 19 5 4 1 0 0 Placebo 21 11 10 6 2 1 1 Subject Accounting between analysis populations ⇓ Diagnosed as HIV+ at or before week 12 visit Discontinued before confirmatory visit Received ART before confirmatory visit Received non-study vaccine Primary dataset reviewed by DSMB 8 07-Nov-2007 HIV Incidence: Ad5 ≤ 200 Personyears of Follow-up** 619 622 Incidence of HIV Infection 3.07 1.77 Population Per-Protocol Group Vaccine Placebo N* 672 691 n 19 11 95% CI (1.85, 4.79) (0.88, 3.16) 1 tailed p-value = 0.949 (for VEINF > 0) MITT Vaccine Placebo 741 762 24 21 822 836 2.92 2.51 (1.87, 4.34) (1.56, 3.84) 1-tailed p-value = 0.743 (for VEINF > 0) *N=Number in respective analysis population **For the per-protocol population, follow-up time is defined as number of days from the day of the Week 12 visit to the last day of study follow-up for uninfected subjects and to the day of diagnosis for infected subjects. For the MITT population, follow-up time is defined as number of days from the day of vaccination to the last day of study follow-up for uninfected subjects and to the day of diagnosis for infected subjects. 9 07-Nov-2007 Viral Load Set-Points: Ad5 ≤ 200 Per-protocol G. Mean (c/mL): Mean (log10 c/mL): n: 1000000 MITT G. Mean (c/mL): Mean (log10 c/mL): n: 1000000 39557 4.6 19 37179 4.57 10 40903 4.61 24 25862 4.41 20 300000 300000 Viral load setpoint (copies/mL) 100000 30000 Viral load setpoint (copies/mL) Vaccine Male G. Mean (copies/mL) 100000 30000 10000 10000 3000 3000 1000 <400 1000 <400 Placebo Vaccine Male G. Mean (copies/mL) Placebo 1-tailed p-value = 0.528 (for VEVL>0) There was 1 female infection: VLS = 20,207 c/mL (4.31 log10 c/mL) 1-tailed p-value = 0.656 (for VEVL>0) 10 07-Nov-2007 Futility declared • Futility cutoffs met at first interim analysis – p-value > 0.50 (1-tailed) for EACH endpoint • Trend favored placebo for each endpoint – Strongly suggests the vaccine neither prevents HIV infection nor reduces the amount of virus in those who became infected with HIV • Based on these results, the DSMB recommended that – No further injections be administered in the trial – Volunteers be encouraged to return for all protocol visits and tests so that the investigators can fully evaluate whether there is an increased risk of infection in vaccine recipients over time – The trial Oversight Committee determine the appropriate steps and timing of release of trial results to volunteers, investigators, those conducting related trials, relevant 11 agencies, and the public 07-Nov-2007 Additional analyses • Since futility had been declared by DSMB, the team felt it was justifiable to proceed (cautiously) with post-hoc analyses – Since there has only been 1 female case, all subsequent analyses will present males only – Additional analyses will focus on MITT population • Most conservative approach since does not exclude any postrandomization cases – Analyses in other Ad5 strata – Analyses of infection endpoint which include additional cases accrued since cut-off for first interim analysis – Focus on deciphering the reasons for lack of efficacy and the implications this has on broader population All analyses which follow are post-hoc 12 07-Nov-2007 Necessary steps prior to additional analyses • Verify data integrity on additional cases – Not all data was included in frozen file used for interim analysis • Complete adjudication process of additional cases • Prioritize and complete additional lab assays – HIV diagnostic testing on additional cases – Key immunological assays • Design, test, implement and review the programming and output of multiple, complex post-hoc statistical analyses 13 07-Nov-2007 Cases included in additional analyses (Males only) Ad5 ≤ 200 male MITT cases included in 1st interim analysis* Additional Ad5 ≤ 200 male MITT cases accrued through Oct 17, 2007 Total Ad5 > 200 male MITT cases accrued through Oct 17, 2007 Total male cases accrued through Oct 17, 2007 44 8 30 82 *Primary dataset reviewed by DSMB, excluding the 1 female infection 14 07-Nov-2007 Cumulative Number of HIV Infections: MITT population (males), Ad5 ≤ 200 30 Cumulative number of HIV infections (events) 28 Vaccine 25 24 Placebo 20 15 10 5 0 0 10 20 30 40 50 60 70 80 90 100 Time to event (weeks) 1-tailed p-value = 0.322 (for VEINF ≠ 0) Cases accrued as of Oct 17, 2007 2-tailed p-value = 0.581 (for VEINF ≠ 0) 15 07-Nov-2007 Cumulative Number of HIV Infections: MITT population (males) Overall 60 Ad5 > 200 30 Cumulative number of HIV infections (events) Cumulative number of HIV infections (events) 55 50 45 40 35 30 25 20 15 10 5 0 0 10 20 30 40 50 60 70 80 90 100 49 Vaccine 25 20 21 Vaccine 33 Placebo 15 10 9 Placebo 5 0 0 10 20 30 40 50 60 70 80 90 100 Time to event (weeks) Time to event (weeks) 1-tailed p-value = 0.044 (for VEINF ≠ 0) 2-tailed p-value = 0.077 (for VEINF ≠ 0) Cases accrued as of Oct 17, 2007 1-tailed p-value = 0.020 (for VEINF ≠ 0) 2-tailed p-value = 0.029 (for VEINF ≠ 0) 16 07-Nov-2007 Cumulative Number of HIV Infections: MITT population (males) 30 Cumulative number of HIV infections (events) Cumulative number of HIV infections (events) Ad5 ≤ 18 20 Placebo 20 Vaccine 30 18 < Ad5 ≤ 200 25 25 20 20 15 15 10 10 8 Vaccine 5 5 4 Placebo 0 0 10 20 30 40 50 60 70 80 90 100 0 0 30 10 20 30 40 50 60 70 80 90 100 30 Time to event (weeks) Cumulative number of HIV infections (events) Time to event (weeks) Cumulative number of HIV infections (events) 25 200 < Ad5 ≤ 1000 25 Ad5 > 1000 20 20 15 14 Vaccine 15 10 10 7 Placebo 5 7 Vaccine 5 0 0 10 20 30 40 50 60 70 80 90 100 0 0 10 2 Placebo 20 30 40 50 60 70 80 90 100 Time to event (weeks) Time to event (weeks) Cases accrued as of Oct 17, 2007 17 07-Nov-2007 Incidence (95% CI) of HIV Infection MITT population (males) Baseline Ad5 titer ≤ 18 19-200 201-1000 > 1000 ≤ 18 > 18 ≤ 200 > 200 Overall Vaccine V 4.0 (2.5, 6.3) 4.4 (1.9, 8.8) 6.1 (3.3, 10.2) 4.4 (1.8, 9.1) 4.0 (2.5, 6.3) 5.1 (3.4, 7.3) 4.2 (2.8, 6.0) 5.4 (3.3, 8.2) 4.6 (3.4, 6.1) Placebo P 4.0 (2.5, 6.2) 2.2 (0.6, 5.5) 3.0 (1.2, 6.2) 1.2 (0.2, 4.5) Relative Incidence (V:P) 1.0 (0.5, 2.0) 2.1 (0.6, 9.3) 2.0 (0.8, 5.9) 3.5 (0.7, 35.0) 1.0 (0.5, 2.0) 4.0 (2.5, 6.2) 2.2 (1.2, 3.8) 3.5 (2.3, 5.2) 2.3 (1.0, 4.3) 3.1 (2.1, 4.3) 2.3 (1.1, 4.7) 1.2 (0.7, 2.1) 2.4 (1.0, 5.8) 1.5 (0.9, 2.4) 18 is the LOQ for the Ad5 titer assay; includes all HIV cases thru Oct 17, 2007 18 07-Nov-2007 Relative Incidence of HIV infection (Vaccine:Placebo) MITT population (males) RI: 95% CI: 10 9 Relative Incidence (Vaccine:Placebo) 8 7 6 5 4 3 2 1 0 <=18 19-200 201-1000 >1000 1.01 ( 0.51 , 1.97 ) 2.06 ( 0.55 , 9.33 ) 2.01 ( 0.76 , 5.88 ) 3.54 ( 0.67 , 34.96 ) Baseline Ad5 titer # Events [# Risk, Mean follow-up in weeks] Vaccine: 20 [ 382 , 67.2 ] 8 [ 140 , 66.9 ] 14 [ 229 , 52.5 ] Placebo: 20 [ 394 , 65.7 ] 4 [ 142 , 67.7 ] 7 [ 229 , 52.7 ] 7 [ 163 , 50.7 ] 2 [ 157 , 53.3 ] 19 07-Nov-2007 Relative Incidence of HIV infection (Vaccine:Placebo) MITT population (males) RI: 95% CI: 6 Relative Incidence (Vaccine:Placebo) 1.01 ( 0.51 , 1.97 ) 2.26 ( 1.14 , 4.74 ) RI: 95% CI: 6 Relative Incidence (Vaccine:Placebo) 1.18 ( 0.66 , 2.13 ) 2.35 ( 1.03 , 5.83 ) 5 5 4 4 3 3 2 2 1 1 <=18 >18 <=200 >200 Baseline Ad5 titer # Events [# Risk, Mean follow-up in weeks] Vaccine: 20 [ 382 , 67.2 ] 29 [ 532 , 55.7 ] Placebo: 20 [ 394 , 65.7 ] 13 [ 528 , 56.9 ] Baseline Ad5 titer # Events [# Risk, Mean follow-up in weeks] Vaccine: 28 [ 522 , 67.1 ] 21 [ 392 , 51.7 ] Placebo: 24 [ 536 , 66.2 ] 9 [ 386 , 52.9 ] 20 07-Nov-2007 Viral Load Endpoint 21 07-Nov-2007 Summary of VL Setpoint: MITT population (males) # Events: GM: 1000000 25 41527 21 26696 21 19070 9 89810 # Events: GM: 1000000 46 29109 30 38416 VL Setpoint (copies/mL) 100000 VL Setpoint (copies/mL) 100000 10000 10000 1000 1000 <400 <400 Vaccine Placebo Vaccine Placebo <=200 Vaccine Placebo >200 Overall For subjects with viral load setpoint data available as of Oct 17, 2007. 22 07-Nov-2007 Summary of Efficacy Results: MITT population (males) HIV Infection Endpoint HIV Case split rate(%/yr) N 1058 778 1836 V 28 21 49 P 24 9 33 V 4.2 5.4 4.6 P 3.5 2.3 3.1 Viral Load Endpoint log10 copies/ml V 4.62 4.28 4.46 P 4.43 4.95 4.58 Baseline Ad5 ≤ 200 > 200 Overall V = vaccine, P = placebo Data as of October 17, 2007 23 07-Nov-2007 Immunogenicity results: Ad5 ≤ 200 • Primary immunogenicity measure is unfractionated γ-interferon ELISPOT – ELISPOT responder: ≥ 55 SFC/10^6 PBMC and ≥ 4fold over negative control (mock) • PBMC collected at Week 8, Week 30, Week 52 and Week 104 in all subjects • Week 8 ELISPOT was run in “real-time” on a randomly selected 25% subset of all subjects and on all cases after they were identified – Other data currently being generated 24 07-Nov-2007 Week 8 ELISPOT Responses % Responders Cases <200 N Gag Pol Nef >200 N Gag Pol Nef 19 74% 63% 74% 13 46% 38% 46% Non-cases 143 76% 73% 70% 173 54% 47% 51% GM Elispot Cases 19 354 627 327 13 181 296 149 Non-cases 143 260 463 237 173 168 241 163 ELISPOT responder: ≥ 55 SFC/10^6 PBMC and ≥ 4-fold over negative control (mock) GM ELISPOT is for all subjects Summaries based on 25% random subset of all subjects and all PP cases 25 07-Nov-2007 Summary • Study design and execution allowed timely assessment of both primary endpoints • There was no evidence that vaccination prevented infection or lowered viral setpoint • There were more infections in vaccinees than placebo recipients – This trend was more pronounced in participants with high baseline Ad5 titers • Lack of efficacy did not appear to be explained by sub-optimal immune responses in vaccinees 26 07-Nov-2007

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