IMPAACT P 1093 Phase III MultiCenter OpenLabel Pharmacokinetic

IMPAACT P 1093 Phase I/II, Multi-Center, Open-Label Pharmacokinetic, Safety, Tolerability and Antiviral Activity of Dolutegravir (GSK 1349572), a Novel Integrase Inhibitor, in Combination Regimens in HIV-1 Infected Infants, Children and Adolescents

IMPAACT P 1093 Introduction and Overview • Rationale: Unmet ARV needs remain in pediatric populations, especially those with extensive resistance, drug intolerance or toxicity. Results from P 1093 will support regulatory filing for dolutegravir (GSK 1349572) use in pediatrics • Hypothesis: Dolutegravir (GSK 1349572), a novel integrase inhibitor, will be well tolerated with acceptable safety profile and ARV activity when used with an optimized background regimen in HIV infected pediatric populations

Primary Objectives • To select a dolutegravir dose for chronic dosing in infants, children and adolescents that achieves similar exposure to the dolutegravir adult dose selected from the Phase IIb clinical trial in ART-naïve adult subjects • To determine the short term and long term safety and tolerability of dolutegravir in HIV-1 infected infants, children and adolescents at 24 and 48 weeks • To evaluate the steady-state pharmacokinetics of dolutegravir in combination with other antiretrovirals (OBT) in treatment-experienced and naïve HIV-1 infected infants, children and adolescents and to determine the dose of GSK 1349572 that achieves a targeted AUC 24 (primary PK endpoint) and C 24 h (secondary PK endpoint) in this population

Secondary Objectives • To evaluate the antiviral activity of dolutegravir in combination with an OBT by measuring virologic response in infants, children and adolescents at weeks 24 and 48 • To evaluate the effect on immunologic response from baseline to weeks 24 and 48 • To assess changes in HIV-1 genotype and phenotype to dolutegravir and other components of the OBT in subjects experiencing virologic failure • To determine the extended long term ( ≥ 48 weeks) safety and tolerability of dolutegravir in HIV-1 infected infants, children and adolescents

IMPAACT P 1093 • DESIGN: Phase I/II, multi-center, open-label, non-comparative study • SITES: International and US • SAMPLE SIZE: Approximately 160 infants, children and adolescents to provide 100 evaluable subjects • POPULATION: HIV-1 infected children and adolescents ages ≥ 4 weeks to <18 years of age • TREATMENT DURATION: 48 weeks plus long term safety follow-up (with drug provided) until available, no benefit, patient endpoint or discontinuation of dolutegravir development • Two stages per cohort – Stage I: At least 10 subjects in each age cohort – Stage II: Opens after dose/safety criteria met in Stage I; 12 subjects per cohort (except cohort IIB)

Age Cohorts • • • Cohort I: Adolescents ≥ 12 to <18 years of age (Tablets: COMPLETED) Cohort IIA: Children ≥ 6 to <12 years of age (Tablets: Stage I COMPLETED) Stage II is ongoing Cohort IIB: Children ≥ 6 to <12 years of age (Pediatric formulation: oral suspension) Cohort III: Children ≥ 2 to <6 years of age (Pediatric formulation: oral suspension) Cohort IV: Children ≥ 6 months to <2 years (Pediatric formulation: oral suspension) Cohort V: Infants >4 weeks to <6 months (Pediatric formulation: oral suspension)

Inclusion Criteria – RNA ≥ 1, 000 copies/ml at screening – ARV treatment experienced (includes those who have received therapy to interrupt maternal-infant transmission) subjects currently on no ART: • Must be off treatment ≥ 4 weeks AND • Must have HIV-1 RNA viral load greater than 1, 000 copies/m. L of plasma at screening – ARV treatment experienced subjects currently on ART: • Must be on an unchanged, failing therapeutic regimen for within 8 to 12 weeks prior to screening (≤ 1 log drop in HIV-1 RNA within the previous 8 to 12 weeks of screening) AND • Must have HIV-1 RNA viral load greater than 1, 000 copies/m. L of plasma at screening – Able to swallow assigned study medication (do to crush or dissolve) – OBT: > 2 ARVs, one must be active (according to genotype) – Female of childbearing potential, must be on 2 birth control methods while on study and for 2 weeks after stopping study drug

Exclusion Criteria • Active AIDS defining opportunistic infection • Known ≥ Grade 3 of laboratory toxicities, 30 days prior to entry; Hb, neutrophil count, platelets, AST, ALT, lipase, creatinine • Known ≥ 4 laboratory toxicity, 30 days prior to entry • Liver toxicity; ALT> 3 x ULN & Direct bilirubin > 2 x ULN • History of malignancy • Clinical evidence of pancreatitis • Use of disallowed medications at the time of screening (section 4. 32) • Integrase inhibitor experienced (incl mother prior to delivery) • Pregnancy • Active TB /disease and/or requirement for treatment that includes rifampin at the time of the screening visit. However, subjects who need rifampin treatment while on dolutegravir will be allowed to continue in P 1093 provided the dolutegravir dose is adjusted according to protocol section 6. 18.

Dolutegravir Background

Dolutegravir: Key Characteristics • Only once daily, unboosted INI – Long plasma half life (~15 hours) – Consistent pharmacokinetics – Clear and predictable exposure-response relationship • Favorable drug interaction profile – Primarily metabolized via glucuronidation with minor CYP 3 A 4 component • Virology: High potency in a low mg, – – 10 day monotherapy: Decrease VL=2. 5 log copies/ml; 7/10 with VL<50 ≥ 88% undetectable at Week 48 (vs. 82% with EFV) in SPRING-1 Limited cross resistance to RAL in vitro and in vivo (VIKING) Promise for higher barrier to resistance • Excellent tolerability through 48 weeks in trt-naïve (SPRING-1) and 24 weeks in trt-exp (VIKING) populations

SPRING-1: Rapid and Robust Antiviral Activity: Week 48 Efficacy Analysis (%<50 copies/m. L) Proportion (%) <50 copies/m. L (TLOVR) 91% 90% 88% 82% GSK 572 10 mg GSK 572 25 mg GSK 572 50 mg EFV 600 mg 11

SPRING-1 Common Adverse Events Preferred Term 12 Nausea Diarrhea Nasopharyngitis GSK 572 10 mg (N=53) 10 (19%) 5 (9%) 7 (13%) GSK 572 25 mg (N=51) 7 (14%) 6 (12%) 5 (10%) GSK 572 50 mg (N=51) 6 (12%) 7 (14%) 6 (12%) GSK 572 Subtotal (N=155) 23 (15%) 18 (12%) EFV 600 mg (N=50) 5 (10%) 7 (14%) 5 (10%) Headache 6 (11%) 7 (14%) 20 (13%) 1 (2%) Dizziness 2 (4%) 3 (6%) 7 (5%) 11 (22%) Insomnia 0 5 (10%) 10 (6%) 5 (10%) Pyrexia Cough Influenza Rash Bronchitis Fatigue Back pain Sinusitis Depression Respiratory tract infection 5 (9%) 4 (8%) 2 (4%) 1 (2%) 2 (4%) 3 (6%) 5 (10%) 2 (4%) 3 (6%) 2 (4%) 1 (2%) 3 (6%) 1 (2%) 2 (4%) 4 (8%) 3 (6%) 1 (2%) 2 (4%) 10 (6%) 11 (7%) 10 (6%) 7 (5%) 8 (5%) 6 (4%) 5 (3%) 6 (4%) 4 (8%) 2 (4%) 5 (10%) 3 (6%) 4 (8%) 2 (4%) Upper respiratory tract infection Abdominal pain 1 (2%) 3 (6%) 1 (2%) 5 (10%) 2 (4%) 7 (5%) 6 (4%) 1 (2%) Abdominal pain upper Abnormal dreams 4 (8%) 1 (2%) 2 (4%) 0 0 6 (4%) 3 (2%) 1 (2%) 4 (8%) Asthenia 3 (6%) 1 (2%) 7 (5%) 0 Vomiting Dyspepsia Pharyngitis Vertigo 3 (6%) 0 2 (4%) 3 (6%) 2 (4%) 0 0 2 (4%) 6 (4%) 4 (3%) 1 (2%) 2 (4%)

SPRING-1: Creatinine, CRT Clearance GSK 572 10 mg (N=53) GSK 572 25 mg (N=51) GSK 572 50 mg (N=51) GSK 572 Subtotal (N=155) EFV 600 mg (N=50) CRT Maximum Treatment Emergent Toxicity Grade 1 Grade 2 0 0 3 (6%) 1 (2%) 0 0 3 (2%) 1 (<1%) 0 0 Creatinine (µmol/L) Baseline Week 1 Change from Baseline (SD) Week 16 Change from Baseline (SD) Week 24 Change from Baseline (SD) Week 48 Change from Baseline (SD) 82. 0 8. 5 (8. 04) 8. 1 (8. 83) 4. 4 (9. 15) 2. 7 (9. 16) 88. 1 9. 0 (8. 85) 7. 0 (9. 77) 5. 1 (11. 91) 1. 1 (11. 38) 82. 2 11. 9 (6. 79) 12. 4 (9. 39) 10. 6 (7. 75) 6. 3 (7. 67) 84. 1 9. 8 (8. 04) 9. 1 (9. 55)* 6. 6 (10. 08)** 3. 4 (9. 69)*** 79. 5 1. 1 (7. 79) 0. 8 (6. 99) -2. 0 (10. 08) -6. 0 (10. 19) Creatinine Clearance - Cockcroft-Gault (m. L/min) Baseline Week 24 Change from Baseline (SD) Week 48 Change from Baseline (SD) 116. 0 -5. 0 (13. 55) -2. 5 (13. 62) 115. 8 -4. 5 (16. 77) -1. 3 (14. 31) 118. 6 -11. 2 (9. 42) -8. 0 (10. 54) 13 * p<0. 001 vs. EFV; ***p<0. 001 vs. EFV; by NRTI: TVD: p<0. 001 vs. EFV KVX: p=0. 021 vs. EFV KVX: p=0. 115 vs. EFV KVX: p=0. 128 vs. EFV 118. 5 4. 7 (16. 25) 13. 7 (28. 02)

SPRING 1: Safety Summary • Dolutegravir was well-tolerated across all doses studied. • No significant dose-dependent trends in safety parameters. • More cases of headache were reported across the DTG treatment groups (13%) than in the EFV control (2%); nausea – DTG (15%), EFV (10%). • Small, non-progressive increases in serum creatinine have been observed across the dolutegravir doses in this study. In vitro data supports OCT 2 inhibition; iohexol study in healthy subjects shows no effect on GFR or renal plasma blood flow • Urine protein changes noted on DTG – not time or dose-dependent, resolve independently, confounded by other conditions – Direct assessments in peds and ph 3 studies

Dolutegravir Pharmacokinetics

Predictable PK/PD Relationship C (μg/m. L) Day 11 log 10 VL Change from Baseline 0. 0 -3. 5 0. 2 0. 4 0. 6 0. 8 1. 0 1. 2 1. 4 -3 -2. 5 -2 -1. 5 -1 -0. 5 Placebo 2 mg QD 0 10 mg QD 0. 5 50 mg QD 1 Model Fit, Emax= -2. 6, EC 50= 0. 036μg/m. L Song I, et al. IAS 2009, Cape Town, abstract WEPEB 250. Well characterized relationship between trough concentration and viral load decrease in 10 -day monotherapy observed

Drug-drug Interactions • Dolutegravir can be given with most ART agents without dose adjustment or contraindication • Dolutegravir can be given without food restrictions • Exceptions (due to significant reduction in dolutegravir exposure) – Disallowed concurrent medications: • Etravirine (without PI/r) • Rilpivirine – Concurrent medications with caution: • GSK 1349572 should be taken 2 hours before or 6 hours after antacid products containing divalent cations (e. g. , Tums, Maalox) Please refer to Section 4. 3 in the protocol for complete list of precautionary and disallowed medications at various stages of the study

Mean (SD) plasma DTG concentration (ng/m. L) PK Result: DTG Exposure in Cohort 1 (12 to <18 y) is Similar to Adults IAC 2012 5000 PK parameter, mean (CV%) 4500 AUC(0 -24) 4000 Cohort 1 46. 0 µg*h/m. L (43%) C 24 0. 90 µg/m. L (58%) 3500 3000 P 1093 Cohort 1 Stage 1 2500 2000 Adult 1500 1000 500 0 0 5 10 Time (h) 15 20 25 DTG exposure in Cohort 1 (12 to <18 y) achieved target exposure for both AUC(0 -24) (37 -67 µg*h/m. L) and C 24 (0. 77 -2. 26 µg/m. L) Adult profile is based on pooled data from Study ING 111521 and SPRING-1: Min et al. AIDS. 2011; 25(14): 1737 -45. Stellbrink et al. CROI 2012; Seattle, WA. Abstract K-1002. van Lunzen et al. Lancet Infect Dis. 2012; 12(2): 111 -118.

PK Result: DTG Exposure in Cohort IIA (6 to <12 years) CROI 2014 PK Parameter Geometric Mean (CV %) AUC 24 50. 46 µg*h/m. L (63%) C 24 0. 92 µg/m. L (89%)

Data From P 1093 – CROI 2014 Dose and Safety at Week 48 • Dose n=23 • 50 mg for ≥ 40 kg 19 (82. 6%) • 35 mg for 30 -< 40 kg 4 (17. 4%) • DTG was generally well tolerated – No discontinuations due to adverse events – No DTG-related AE – Two participants with unrelated grade 3 laboratory abnormality • Unconjugated bilirubin elevation associated with atazanavir • Asymptomatic lipase elevation – No trends in lab abnormalities

P 1093: Efficacy: Percent of Patients (95% CI) with HIV RNA<400 c/m. L or ≥ 1 Log 10 Decline from Baseline (Cohort I - All Treated): ITT Approach 74%

Efficacy: Percent of Patients (95% CI) with HIV RNA<400 c/m. L or ≥ 1 Log 10 Decline from Baseline (Cohort IIA - All Treated): ITT Approach 82%

P 1093: CD 4+ Counts Median Changes (IQR) from Baseline through Week 48 (Cohort I): As-treated Approach

P 1093 Study Visits

Age Cohorts • Cohort I: Adolescents ≥ 12 to <18 years of age (Tablets: COMPLETED) • Cohort IIA: Children ≥ 6 to <12 years of age (Tablets: Stage IA COMPLETED) Cohort IIB: Children ≥ 6 to <12 years of age (Pediatric formulation: oral suspension) Cohort III: Children ≥ 2 to <6 years of age (Pediatric formulation: oral suspension) Cohort IV: Children ≥ 6 months to <2 years (Pediatric formulation: oral suspension) Cohort V: Infants >4 weeks to <6 months (Pediatric formulation: oral suspension) • •

P 1093 Study Cohorts (similar to P 1066) • Stage I: Mini-cohort: n=4 – Objective: Screen for enrolling full cohort and subsequent mini-cohort – Add dolutegravir to stable, failing regimen or start as monotherapy if not on ARV – Intensive PK (day 5 -10) – Optimized background regimen (>2 ARV, one active) to be initiated after PK (see Table 8 for excluded ARVs) • One active ARV by genotype at screen • Agents chosen based upon ARV history, prior genotype/pheno) • Send proposed OBT to Team for approval – Team evaluation: Intensive PK and 4 week safety and tolerability – If acceptable, then enroll Full Cohort and start enrollment of subsequent mini-cohort – If not, new mini-cohort at adjusted dose – Original mini-cohort dose adjusted, too

P 1093: Full Cohort, Stage I • Stage I: Full Cohort (finish enrollment of 10) – Team Evaluation: 4 week PK and safety – If safety/PK results not acceptable, start a new mini-cohort at adjusted dose • Case by case individualized dose adjustment for participants with PK extremes for safety reason (Stage 1 only) – Need to have repeat PKs 5 -14 days post dose mod – if refuse to have PKs, must discontinue 572

P 1093: Stage II • Stage II – Objective: Long term safety, tolerability, efficacy of dolutegravir with optimized background – Dose employed, for each cohort, will be chosen from cohort data from Stage I – No Intensive PK – Population PKs – See Table 8 for permissible optimized background regimen

Disallowed ARVs (Table 9) (to minimize effect of drug: drug interaction) • • • Stage I: Prior to Intensive PK – Efavirenz – Nevirapine – Etravirine Include use in any fixed-dose combination – Atazanavir: unboosted and boosted – Fosamprenavir: unboosted and boosted – Tipranavir/ritonavir – Rilpivirine (includes Complera) Stage I: Before or After Intensive PK and all Stage II participants – Etravarine – Raltegravir – Elvitegravir – Rilpivirine (includes Complera) See list of other disallowed medications – Short course of corticosteroids allowed

Screening Highlights (Appendix I) see Manual of Procedures (MOPS) • • Must contact team for permission to screen Real time HLA B*5701 for those using abacavir in OBR-minimize hypersensitivity reactions Entry within 30 days of screening HIV-1 RNA PCR: Abbott platform Specimen shipping: See Clarification memo Pregnancy testing-screen and q visit – urine or blood Real time genotyping – Must have one fully sensitive ARV to add to OBR

Intensive PKs P 1093 • • • Day 5 -10, 24 hr PK Dosing: observed, as close to 24 hrs post last dose Prior – Document 2 doses prior to intensive PK – Fast: 6 hrs but substantial liquids OK until 4 hrs prior to dosing (water OK throughout) Cancellation/rescheduling: complete within 7 days – Missed dose within 72 hrs – Missed 20 -28 hr window after last dose – Vomiting within 4 hrs post dose – Not fasting – Notify team SHIP REAL TIME

Non-Intensive PKs P 1093 • Population PKs – Study visit windows • Weeks 4, 8, 12, 16: one week window • Weeks 24, 32, 40, 48: two week window – Week 24: 2 samples between 12 -26 hrs post dose • Liver toxicity resulting in study drug interruption – Request to collect 2 ml specimen within 72 hrs after last dose

Virologic Criteria • Virologic Rebound – Confirmed >1. 0 log in RNA over nadir level on 2 consecutive measurements (1 -4 weeks after 1 st suspect specimen) • Virologic Failure – Confirmed decrease of HIV RNA <1. 0 log at, or after, wk 12 OR – Confirmed RNA >400 at >24 weeks on 2 consecutive measurements (1 -4 weeks after 1 st suspect specimen)

Participant Management Virologic • • Notify Team See Appendix I for schedule of events Send specimen for resistance testing Participant options: – Take off study drug, best available treatment • Long Term Follow-up for subjects who discontinue study medication: Appendix IF – Optimize background regimen + dolutegravir • Need approval of P 1093 team – Continue regimen without changes *** Timing of which sample to send for resistance testing determined by above option chosen (see MOPS)

Pharmacy Considerations IMPAACT P 1093 Thucuma Sise, Pharm. D. , BCPS Pharmacist Pharmaceutical Affairs Branch DAIDS/NIAID/NIH

Study Drugs: Formulations • Dolutegravir 10 mg study drug tablet • Dolutegravir 25 mg study drug tablet • Dolutegravir 50 mg study drug tablet • Dolutegravir Pediatric Formulation – oral suspension – Granules must be reconstituted by the site pharmacist

Study Drugs: Duration of Therapy • Stage I: Minimum of 48 weeks Long Term Safety Follow-up: see Section 5. 11 of the protocol. • Stage II: 48 weeks Long Term Safety follow-up: see Section 5. 11 of the protocol.

Study Drugs: Dispensing and Storage • Dolutegravir tablets are to be dispensed by the site pharmacist in the original manufacturer’s container, which contains a desiccant. The desiccant should remain in the container. • Dolutegravir pediatric granules are to be reconstituted by the site pharmacist and must remain in the same amber glass bottle in which the granules were mixed with water. • Please refer to Section 5. 1 “preparation of oral suspension”. • Dolutegravir study drugs must be stored between 15°C– 30°C (59°F – 86°F)

Study Drug: Administration of Tablets • Dolutegravir - study drug - must be administered orally once daily or as twice daily if administered with efavirenz or ritonavir boosted fosamprenavir or tipranavir*** • Dolutegravir tablets must be swallowed whole and may not be crushed or dissolved before administration • Dolutegravir - study drug - can be taken with or without food • Dolutegravir - study drug - should be administered 2 hours before or 6 hours after taking antacid products containing divalent cat ions (e. g. aluminum and magnesium) or iron supplements Please refer to Section 4. 3 in the protocol for complete list of precautionary and disallowed medications at various stages of the study ***If EFV, NVP, FPV/r or TPV/r are part of OBR after the intensive PK the dose of dolutegravir should be given TWICE daily as directed in LOA #2 and per FDA guidance

Study Drugs: Administration of Suspension • The site pharmacist will dispense the pediatric suspension reconstituted from granules • EVERY reconstituted bottle must be shaken EVERY day • Dolutegravir - study drug - must be administered orally once daily*** • Dolutegravir - study drug - can be taken with or without food • Dolutegravir - study drug - should be administered 2 hours before or 6 hours after taking antacid products containing divalent cat ions (e. g. aluminum and magnesium) or iron supplements Disperse directly to the mouth Dosing instructions and an oral dispenser to measure the child’s dose will be provided by the pharmacist (See Appendix IV for directions for parent or caregiver) • • ***If EFV, NVP, FPV/r or TPV/r are part of OBR after the intensive PK the dose of dolutegravir should be given TWICE daily as directed in LOA #2 and per FDA guidance

Study Drug: Ordering and Shipping • Site Protocol Registered • Ordering instructions will be sent to the site pharmacist from the Clinical Research Products Management Center (CRPMC) • Upon confirmation of site protocol registration • Receipt of order at the CRPMC • Study product packaged and shipped to the site pharmacist • Site pharmacists will receive study product shipped at appropriate temperature

Study Drugs: Management and Accountability • Accountability records must be kept by the Pharmacist • All unused study product must be handled as outlined in the manual “Pharmacy Guidelines and Instructions for DAIDS Clinical Trials Networks” at study completion or termination.

Communication Among Site Study Staff • A completed prescription signed by an authorized prescriber should include pertinent information including the subject’s weight (kg), Cohort, target mg/kg, Specific Dosing Table in Appendix 3 of the protocol to use for initial dosing. • A new prescription must be provided to the site pharmacist for adjusted dosing.

P 1093 AE Reporting

P 1093 Adverse Event Reporting • Serious Adverse Events (SAE) is the reporting category to be used for this study. • The study agent for which expedited reporting are required is dolutegravir

SAE Definition • Results in Death, • Is life-threatening, • Requires inpatient hospitalization or prolongation of existing hospitalization, • Results in persistent or significant disability or incapacity, • Is a congenital anomaly/ birth defect, • Is an important medical event that may not be immediately life threatening or result in death or hospitalization but may jeopardize the patient or may require intervention to prevent one of the other outcomes listed in the definition above

P 1093 SAE Reporting • In addition to SAE, this protocol also requires reporting of the following in an expedited manner: – ALL GRADE 4 toxicities – All Pregnancies – Hy’s law liver toxicities – Abacavir (ABC) hypersensitivity (HSR) – All pregnancy complications, elective terminations and spontaneous abortions

Abacavir Hypersensitivity Reactions • ALL cases of Abacavir Hypersensitivity reactions, MUST be reported as an EAE. • In addition, the ABC HSR CRF must be completed and submitted to the DMC within one week of the onset of the hypersensitivity reaction.

Liver Toxicities • HY’s law, (section 6. 13) • If any of the liver chemistry stopping criteria are met: – Immediately withhold study drug – Report to study team by email within 24 hours of learning of the event to – Impaact. teamp 1093@fstrf. org – Report Hy’s law liver toxicity to RSC as an EAE – Complete and submit the liver toxicity CRFs within 1 week to the DMC.

Assessment • AEs are assessed for: – – Seriousness Relationship Expectedness Severity • Site Level: Study physician listed on the 1572/Io. R (Investigator of Record) Agreement is responsible for the assessment of AEs • Sponsor Level: DAIDS MOs provide secondary review 50

Serious versus Severe Seriousness is NOT the same as Severity Seriousness Severity Based on outcome of the AE and is a factor in determining reportability (regulatory definition) Based on the intensity of the AE and is not a factor in determining reportability (clinical descriptor) Determined using the SAE criteria Determined using the DAIDS grading table 51

Site Investigator Signature • A site physician investigator or sub-investigator listed on the 1572 or the Io. R Agreement must: – Review and verify the completed report for accuracy and completeness and – Sign the report • This physician makes the site’s final assessment of the relationship to study agent(s) 52

P 1093 CRFs / Data Management

The EVW 0302 is being updated to allow patients who are taking TABLETS to skip over the SUSPENSION questions. Until the update is released, please enter (-1) in the questions collecting suspension feel and ease to swallow? Propose the Logical Check in RESOLVE and the DMC will approve the check.

GSK 1349572 (Dolutegravir) modifications are reported in question #1.

Modifications to the background ARV regimen are reported in question #2. Modifications to non-ARVs are reported in question #4 along with the reason for the modification.

P 1093 LABORATORY TESTS LPC SHIPPING

Evaluation DMC Test Code Tests CRF # Hematology N/A CBC (to include platelet and differential) PE 6811 Chemistry N/A Chemistry-Electrolytes (sodium, potassium, HCO 3), glucose, creatinine, lipase, phosphorus, and LFTs (total bilirubin, indirect bilirubin, alkaline phosphatase, AST, , and albumin. ) PE 6816 Lipid Profile N/A Lipid profile- triglycerides, cholesterol, HDL, LDL PE 6816 Urinalysis N/A Urinalysis (dipstick only) PE 0811

Section 3: Specimen Processing & Shipping Instructions Evaluation Abbott Real Time HIV 1 Tube Type Special Collection Notes CRF # DMC Test Code 3 m. L EDTA Can be performed F 3006 (and Spin blood at 800 xg at any local F 3109 if for 10 mins; remove CLIA/VQA results are plasma and respin at approved lab. not reported 800 xg for 10 min. through Freeze ≥ 2 x 0. 8 ml LDMS) aliquots at – 70°C or lower. RNAHIVRU Processing Shipping To be updated

2 m. L EDTA N/A GENOHIV Resistance testing (Genotyping) BASELINE Resistance Testing (Phenotyping) 4 m. L EDTA F 3006 N/A F 3006 PHENOHIV NOTE: To be collected only if sufficient blood volume Resistance Testing (genotyping) Virologic Failure 2 ml EDTA N/A F 3006 GENOHIV Spin blood at 800 xg for 10 mins; remove plasma and respin at 800 xg for 10 min. Save all plasma in one aliquot and freeze at – 70°C or lower. Specimens should be shipped in Real Time. . Spin blood at 800 xg for 10 mins; remove plasma and respin at 800 xg for 10 min. Save all plasma in two aliquots and freeze at – 70°C or lower. US sites: Store locally and ship to Monogram Biosciences when requested. Spin blood at 800 xg for 10 mins; remove plasma and respin at 800 xg for 10 min. Save all plasma in one aliquot and freeze at – 70°C or lower. Specimens should be shipped in Real Time. . US Sites: Ship to UNC. Thailand – PHPT lab, Chiang Mai, Thailand; Africa sites: ship to Contract Lab Services, Johannesburg, SA; South America Sites: Fio. Cruz, Brazil. International sites store locally and ship to BRI when requested. (NOTE: Specimens to be forwarded from BRI to Monogram) US Sites: Ship to UNC. International Sites: Specimens should be shipped in real time to BRI. (Specimens to be forwarded from BRI to UNC. )

HLA B-5701 Typing NOTE: Sample ONLY should be drawn on subjects whose physician plans to start ABC as part of their OBR, and where ABC hypersensitivity screening is not done as part of standard of care. 1 m. L EDTA M/C Ratio Assay Microalbumin/creatinine ratio (mcg/mg creatinine 4 m. L Urine N/A F 3006 HLAIPCR N/A SPW 0427 Spin blood at 800 xg for 10 mins; remove plasma and re-spin at 800 xg for 10 min. Save all plasma in one aliquot and freeze at – 70°C or lower. Ship real time to BRI as pass through. (NOTE: BRI will forward all samples real time to final destination, Quest. ) Aliquot urine into 2 aliquots at 2. 0 m. L each. South Africa sites to ship to CLS Freeze at – 20°C or lower. Lymphocyte subsets. CD 3/CD 4, CD 3/CD 8 cell counts and percentages 1 m. L EDTA Send to local IQA or CLIA certified (or equivalent) lab, ambient. The same lab must be used throughout the study PBMCs/Plasma for storage for future studies. 4. 5 m. L EDTA Send to IMPAACT processing lab ambient LBW 0054 None CD 4 CD 8 Dual platform labs only must also have a WBC and diff. F 3006 Spin blood at 400 xg for 10 mins; remove plasma and respin at 800 xg for 10 min. Freeze 2 x 1 ml aliquots at – 70°C or lower STORVIR Freeze all PBMCs viably in 5 X 106 aliquots. N/A NIAID Sites: Ship to BRI as directed. NICHD Sites: Ship to Fisher as directed.

Intensive Pharmacokinetics 2 m. L EDTA per time point for Cohorts I, II and II 1 m. L EDTA per time point for Cohorts IV and V. Subjects must be fasted for 6 hours prior to dosing (see Appendix I footnote #11 for additional info. ) Time points: Pre-dose, 1, 2, 3, 4, 6, 8, and 24 hours post dosing. PKW 0290 Centrifuge blood within one hour of PKINT collection at 1000 x g Transfer all plasma to one pre-labeled 2 ml cryovial, and freeze immediately after processing at -70 C or lower for 10 minutes at 0 -5 o. C. International sites ship to BRI real time as pass through to UAB. (NOTE: BRI will forward all samples real time to UAB. ) Send to local IMPAACT processing lab on ice. Population 1 m. L EDTA 4 Weeks: Two samples PKW 0291 Centrifuge blood Pharmacokinetics per time collected at pre-dose PKPOP within one hour of point and 2 -4 hours post dose. collection at 1000 x g for 10 minutes at 0 -5 C. 12 Weeks: One sample Transfer all plasma to at any time post dose. one pre-labeled 2 ml 24 Weeks: Two samples between 12 and 26 hours post dose. Send to local IMPAACT processing lab on ice. cryovial and freeze immediately after processing at -70 C or lower. International sites store locally and ship in batches to BRI when requested as pass through to UAB (NOTE: BRI to forward to UAB as requested. )

Questions?