Clinical trial NCT04291508
Conditions: Acute Respiratory Distress Syndrome Critical Illness Respiratory Failure Sepsis
Keywords: ARDS Acetaminophen Vitamin C Sepsis
Sponsor: Massachusetts General Hospital
Start date: 2021-10-13; Completion date: 2023-07-27
Enrollment: 488 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
Prospective multi-center phase 2b randomized placebo-controlled double-blinded interventional platform trial of two different pharmacologic therapies (intravenous Vitamin C or intravenous Acetaminophen) for patients with sepsis-induced hypotension or respiratory failure.

Detailed description:
Hypothesis 1A: Acetaminophen (APAP) or Vitamin C infusion will increase the days alive and free of organ support to day 28.

Hypothesis 1B: APAP or Vitamin C will have a favorable effect on other secondary outcomes including pulmonary and non-pulmonary organ dysfunction and biomarkers of inflammation and endothelial injury

The investigators plan to carry out two multi-center phase 2b randomized double-blinded placebo-controlled trials of two different pharmacologic therapies within a single platform trial.

1. One trial will assess the efficacy of Acetaminophen (1 gram intravenously every 6 hours) for 120 hours in patients with sepsis who have evidence of either hemodynamic or respiratory organ failure.
2. A second trial will assess the efficacy of Vitamin C (50 mg/kg every 6 hours) infused intravenously for 120 hours in patients with sepsis who have evidence of either hemodynamic or respiratory organ failure.

A total of 900 participants who meet all of the inclusion criteria and none of the exclusion criteria, were planned be randomized in a 2:1:2:1 fashion (APAP-Active: APAP-Placebo: Vit C-Active: Vit C-Placebo). The APAP and Vitamin C trials were planned to be resulted separately. With the closure of the Vitamin C arm in June 2022; the study proceeded with the APAP and Placebo arms with a 1:1 randomization scheme. The total sample size for the APAP trial was 447 participants (227 in the active arm and 220 in the placebo arm). The total sample size for the Vitamin C trial was 79 (40 in the active arm and 39 in the placebo arm). The total combined number in the 4 arms of the ASTER trial was 526 (227 APAP active, 220 APAP placebo, 40 Vit C active, 39 Vit C placebo), although a total of only 487 patients were actually randomized (this is due to the 39 pooled placebo patients that appear in both trials).

Arms: arm 1: Patients randomized to the Acetaminophen arm will receive Acetaminophen at the dose of 1 gram (or 15 mg/kg if actual body weight \< 50kg) in 100 ml 5% dextrose in water every 6 hours intravenously for 5 days (20 doses). arm 2: Patients randomized to the Vitamin C arm will receive Vitamin C at the dose of 50 mg/kg in 100 ml 5% dextrose in water every 6 hours intravenously for 5 days (20 doses). Note: This arm is now closed. arm 3: Patients randomized to placebo will receive an identical-appearing intravenous infusion of 100 ml of 5% dextrose in water every 6 hours for 5 days (20 doses). arm 4: Patients randomized to placebo will receive an identical-appearing intravenous infusion of 100 ml of 5% dextrose in water every 6 hours for 5 days (20 doses). Note: This arm is now closed.
Interventions: intervention 1: Intravenous Acetaminophen (room temperature) intervention 2: Intravenous Vitamin C (refrigerated) intervention 3: 5% Dextrose (room temperature) intervention 4: 5% Dextrose refrigerated
Intervention descriptions: intervention 1: Acetaminophen given intravenously at the dose of 1 gram (or 15 mg/kg if patient weighs \< 50 kg) every six hours for 5 days (20 doses) intervention 2: Vitamin C given intravenously at the dose of 50 mg/kg every six hours for 5 days (20 doses) intervention 3: Placebo (identical appearing room temperature 5% dextrose solution) infused every six hours for 5 days (20 doses) intervention 4: Placebo (identical appearing refrigerated 5% dextrose solution) infused every six hours for 5 days (20 doses)

Protocol documents:
SAP_002.pdf: 
ASTER SAP  
pg. 1 
Acetaminophen in Sepsis: Targeted Therapy to Enhance 
Recovery (ASTER) 
Protocol Version: 4.5 April 25, 2023
Statistical Analysis Plan V5.2 August 1, 2023 
Revised per DSMB recommendations September 21, 2023 
Summary of revisions: 
•
Pg 4: Section 1.4.2 Sample Size/Statistical Considerations: Sentence added: Using
clinical trial data from the PETAL Network, the estimated standard deviation of 28-day
organ support free days (obtained from the PETAL Network CLOVERS study) is 8.7
days.
•
Pg 10: Section 2.6 Interim Analysis: References added to the second paragraph:
1] Martin, G.S., et al., The epidemiology of sepsis in the United States from 1979 through 2000. N Engl J
Med, 2003. 348: p. 1546-1554.
[2] Haybittle, J.L., Repeated assessment of results in clinical trials of cancer treatment. Br J Radiol, 1971.
44(526): p. 793-7.
•
Pgs 12 and 13: Safety Outcomes section: References noted in paragraphs 3 and 5
footnoted and placed in Reference section.
•
Page numbering added to document and formatting edits made.
Revisions reviewed and approved by PETAL CCC PI Andrea Foulkes 
ASTER
TRIAL
APAP
SIGN


ASTER SAP  
 
pg. 2 
 
Contents 
1.Trial Summary ......................................................................................................................................... 3 
1.1 Title .................................................................................................................................................... 3 
1.2 Objective ........................................................................................................................................... 3 
1.3 Hypothesis ........................................................................................................................................ 3 
1.4 Study Design .................................................................................................................................... 3 
1.4.1 Treatment Arms ........................................................................................................................ 3 
1.4.2 Sample Size/Statistical Considerations: ............................................................................... 4 
1.5 Inclusion Criteria .............................................................................................................................. 5 
1.6 Exclusion Criteria ............................................................................................................................. 5 
1.7 Primary Endpoint ............................................................................................................................. 6 
1.8 Secondary Endpoints ...................................................................................................................... 6 
1.9 Safety Variables ............................................................................................................................... 7 
2. Endpoints ................................................................................................................................................ 7 
2.1 Primary Outcome ............................................................................................................................. 7 
2.2 Secondary Outcomes ..................................................................................................................... 8 
2.3 Safety Endpoints .............................................................................................................................. 9 
2.4 Predictive Biomarker Analysis ..................................................................................................... 10 
2.5 Statistical Analysis Plan ................................................................................................................ 10 
2.6 Interim Analysis .............................................................................................................................. 10 
3. Sub-Group by Treatment Interaction ................................................................................................ 11 
4. Statistical Methods ............................................................................................................................... 11 
Overview ............................................................................................................................................. 11 
Primary Efficacy Outcomes ................................................................................................................. 11 
Secondary Efficacy Outcomes ............................................................................................................. 11 
Subgroup by Treatment Interactions for the Primary and Secondary Efficacy Outcomes ................. 12 
Safety Outcomes ................................................................................................................................. 12 
Predictive Biomarker Analysis ............................................................................................................. 13 
References .............................................................................................................................................. 14 
Table of Variables.................................................................................................................................... 15 
 
 


ASTER SAP  
 
pg. 3 
 
 
1.Trial Summary 
1.1 Title 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
ASTER is a platform originally designed to conduct two separate trials. In response to external 
trial results, enrollment in the Vitamin C arm of the trial was suspended on June 15, 2022.  The 
Acetaminophen (APAP) arm of the trial was resized and will continue as described in this 
amended protocol. 
1.2 Objective 
Objective: To carry out one multi-center phase 2b randomized double-blinded placebo-
controlled trial of one pharmacologic therapy. The trial will assess the efficacy of 
Acetaminophen (1 gram intravenously every 6 hours) in comparison to placebo for 120 hours 
in patients with sepsis who have evidence of either hemodynamic or respiratory organ failure. 
The placebo infusion (5% dextrose in water, D5W) will match the volume and storage 
temperatures of the acetaminophen (room temperature, dose adjusted for weight below 50 kg) 
1.3 Hypothesis 
Hypothesis: Acetaminophen (APAP) infusion will increase the days alive and free of organ 
support to day 28. 
1.4 Study Design 
Prospective multi-center phase 2b randomized placebo-controlled double-blinded 
interventional trial of intravenous Acetaminophen for patients with sepsis-induced hypotension 
or respiratory failure.   
1. Enrollment Period: Approximately 15 months  
2. Patient Population: Critically ill adults admitted or planned admission to the ICU with 
sepsis and either hemodynamic or respiratory organ failure.  A maximum of 450 
patients will be enrolled  
3. Locations: Emergency Department and ICUs at hospitals participating in the NHLBI 
PETAL Network  
1.4.1 Treatment Arms 
Patients with sepsis and either hemodynamic or respiratory organ failure on enrollment will be 
randomized to one of two treatment arms in a 1:1 fashion.  The two arms will be 
Acetaminophen and Placebo randomized 1:1.  
 


ASTER SAP  
 
pg. 4 
 
 
 
Patients, nurses, research staff, and physicians will be blinded to the treatment assignment. 
The time of randomization will represent time zero.  Study drug will be started within 4 hours of 
randomization. 
 
2. Completion of study drug administration: Study drug administration will occur for a 
total of 120 hours, unless one of the following occurs:  
1. Discharged from the study hospital  
2. Discharge from the ICU  
3. Withdrawal from the study   
4. Death  
5. New AST or ALT elevation ≥ 10 times the upper limit of normal 
1.4.2 Sample Size/Statistical Considerations:  
Using clinical trial data from the PETAL Network, the estimated standard deviation of 28-day 
organ support free days (obtained from the PETAL Network CLOVERS study) is 8.7 days. With 
randomization of 450 total patients (225 Acetaminophen 225 placebo) we will have 85% power 
to detect a difference between groups of 2.5 days in the primary composite outcome of days 
1. Acetaminophen given intravenously at the dose of 1 gram (or 15 mg/kg if patient weighs < 50 
kg) every six hours for 5 days (20 doses) OR 
2. Placebo (identical appearing 5% dextrose solution) infused every six hours for 5  days (20 
doses)  
NOTE: The volume of placebo infusions will be lower in patients <50kg and will be matched to 
Acetaminophen.  
Treatment will continue 
for 120 hours (20 
doses), or discharge 
from the intensive care 
unit, study withdrawal, 
or death, whichever 
comes first. 
First study drug dose  
(APAP or placebo) will be 
considered “Dose 1” and will 
be administered within 4 
hours of randomization.   
Study drug will be infused 
over 30 minutes.  
 
Subsequent doses will be infused 
every six hours (+/- 1 hour) through 
120 hours or 20 doses (whichever 
comes first).  If a dose cannot be 
administered within 3 hours of the 
scheduled time, the dose should be 
skipped.   
Any dose administered outside of the 
+/- 1 hour window or any dose that is 
skipped will trigger a protocol deviation 
Duration 
Subsequent Dosing 
Timing of Dosing 


ASTER SAP  
 
pg. 5 
 
alive and free of organ support (dialysis, assisted ventilation, and vasopressors) to day 28. 
Patients will be analyzed on an intention-to-treat basis based on randomization assignment.  
 
1.5 Inclusion Criteria 
1. Age ≥ 18 years  
2. Sepsis defined as:  
a. Clinical evidence of a known or suspected infection and orders written to administer 
antibiotics  
AND  
b. Hypotension as defined by the need for any vasopressor (and at least 1 liter of fluid 
already administered intravenously for resuscitation) OR respiratory failure defined 
by mechanical ventilation, BIPAP or CPAP at any level, or greater than or equal to 
6 liters/minute of supplemental oxygen (criterion b must be present at the time of 
randomization)  
3. Admitted (or intent to admit) to a study site ICU within 36 hours of presentation to the 
ED or any acute care hospital  
1.6 Exclusion Criteria 
1. No consent/inability to obtain consent from the participant or a legally authorized 
representative  
2. Patient unable to be randomized within 36 hours of presentation to the ED or within 36 
hours of presentation to any acute care hospital  
3. Diagnosis of cirrhosis by medical chart review  
4. Liver transplant recipient   
5. AST or ALT greater than five times the upper limit of normal   
6. Diagnosis of ongoing chronic alcohol use disorder/abuse by chart review; if medical record 
unclear, use Appendix F  
7. Hypersensitivity to Acetaminophen  
8. Patient, surrogate or physician not committed to full support (Exception: a patient will not 
be excluded if he/she would receive all supportive care except for attempts at resuscitation 
from cardiac arrest)  
9. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used 
only for sleep-disordered breathing  
10. Chronic dialysis 
11. Use of home oxygen > 3 liters/min nasal cannula for chronic cardiopulmonary disease  
12. Moribund patient not expected to survive 24 hours   
13. Underlying malignancy or other condition with estimated life expectancy of less than 1 
month  


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pg. 6 
 
14. Pregnant woman, woman of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or woman who is breast feeding  
15. Prisoner  
16. Treating team unwilling to enroll because of intended use of Acetaminophen  
 
1.7 Primary Endpoint 
Primary efficacy variables will be evaluated comparing Acetaminophen vs. placebo The primary 
efficacy variable is days alive and free of organ support (dialysis, assisted ventilation, and 
vasopressors) to day 28.  
Participants will need to be free of all three components (assisted ventilation, vasopressors, new 
renal replacement therapy) to qualify for a day alive and free from organ failures. A patient could 
be free of all organ failures from day 10 through day 22 and then die on day 22 and would 
receive 12 days of credit for this primary outcome.  The components of this outcome (days free 
of dialysis, assisted ventilation, and vasopressors in the overall cohort and in survivors, and 28-
day all-cause all-location mortality will also be reported as secondary outcomes (see below in 
2.1). 
 1.8 Secondary Endpoints 
Secondary efficacy variables will be evaluated comparing Acetaminophen vs. placebo.  
1. 28-day ventilator-free days   
2. 28-day vasopressor-free days   
3. 28-day new renal replacement-free days  
4. 28-day hospital mortality 
5. 28-day ICU free-days  
6. 28-day hospital-free days to discharge home  
7. 28-day duration of ICU stay in survivors and non-survivors   
8. Initiation of assisted ventilation to day 28  
9. Initiation of renal replacement therapy to day 28  
10. Change in Sequential Organ Failure Assessment (SOFA) scores between enrollment 
and study day 7  
11. 90-day hospital mortality (includes death at any healthcare facility that is not the 
equivalent of home for a patient)  
12. Development of ARDS within 7 days of randomization   
13. Change in serum creatinine from enrollment to discharge, death, initiation of dialysis or 
28 days, whichever occurs first   
14. Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline, need for new renal replacement therapy, or death  


ASTER SAP  
 
pg. 7 
 
15. Change in the Radiographic Assessment of Lung Edema (RALE) score from 
enrollment to study day 3 in patients who are receiving assisted ventilation or high flow 
nasal oxygen at baseline.  
16. 90-day all-cause mortality   
1.9 Safety Variables  
The safety monitoring and analyses will be performed separately on the Acetaminophen and 
placebo arms (1 active to 1 placebo randomization ratio). The safety variables will include the 
following:  
1. Hypersensitivity or rash  
2. AST and ALT measured on study days 0, 2-5, and 7 (day 7 measurement can be ±1 day) 
in the APAP-Active/APAP-Placebo arm only.  
3. The plasma levels of Acetaminophen will be obtained as a trough level just prior to 
administration of a study dose on study day 2 (subject must have received at least 5 
doses of study drug before the day 2 samples are obtained) for all patients. After the first 
100 patients have been enrolled who have been randomized to the Acetaminophen arm, 
the trough levels will be reported to the DSMB to determine if they exceed the level of 20 
micrograms/mL, the potential level for hepatotoxicity.  
4. Administration of fluid bolus, new use of vasopressor, or increased dose of vasopressor 
within 120 minutes of study drug infusion.  
5. Incidence of reported adverse events  
2. Endpoints 
2.1 Primary Outcome 
The primary endpoint of this trial is to evaluate the effects of intravenous Acetaminophen 
compared to placebo on the number of days alive and free of assisted ventilation, 
vasopressors and new renal replacement therapy between the treatment arm and the placebo 
group over the first 28 days.   
Note that the primary outcome components to be reported will be:  
1. 28-day all-location all-cause mortality  
2. Days free of assisted ventilation to day 28 in the overall cohort and survivors  
3. Days free of new RRT to day 28 in the overall cohort and survivors  
4. Days free of vasopressors to day 28 in in the overall cohort and survivors  
The primary safety objective is to determine the effect of intravenous Acetaminophen on 
markers of liver injury (AST and ALT) measured on days 2-5, and 7 (day 7 measurement can 
be ±1 day).  


ASTER SAP  
 
pg. 8 
 
2.2 Secondary Outcomes 
We will compare the efficacy of intravenous Acetaminophen to placebo for efficacy endpoints.  
We will compare the safety of Acetaminophen to placebo for safety endpoints.  
1. 28-day ventilator-free days: VFDs depend on both duration of ventilation and 
mortality through study day 28. In participants who survive 28 days, VFD is defined as 
28 minus days of invasive or noninvasive ventilation to day 28.  Duration of ventilation 
is counted from the first study day of assisted breathing through the last day of assisted 
breathing provided the last day is prior to day 28. For participants discharged with 
assisted ventilation (e.g., to LTAC facility) prior to day 28, a phone call will be required 
to assess ventilator and vital status at day 28. Participants discharged prior to day 28 
on unassisted breathing will be assumed to remain on unassisted breathing through 
day 28. Isolated periods of ventilation briefer than 24 hours for surgical procedures and 
ventilation solely for sleep disordered breathing do not count towards duration of 
ventilation. In participants who never require assisted breathing, duration of ventilation 
is zero. Participants who do not survive 28 days will be assigned zero VFD.  
2. 28-day vasopressor-free days: Vasopressor free days to day 28 are defined as the 
number of calendar days between randomization and 28 days later that the patient is 
alive and without the use of vasopressor therapy.  Patients who die prior to day 28 and 
those who receive vasopressor therapy for the entire first 28 days are assigned zero 
vasopressor free days.    
3. 28-day new renal replacement-free days: Renal replacement free days to day 28 are 
defined as the number of calendar days between randomization and 28 days later that 
the patient is alive and without renal replacement therapy. We also follow the “last off” 
method. Patients who died prior to day 28 and those who receive renal replacement 
therapy for the entire first 28 days are assigned zero renal replacement free days.  
4. 28-day hospital mortality  
5. 28-day ICU free days:  ICU free days to day 28 are defined as the number of days 
spent alive and out of the ICU to day 28.   
6. 28-day hospital free days to discharge home: Hospital free days to day 28 are 
defined as 28 days minus the number of days from randomization to discharge home. 
If a patient has not been discharged home prior to study day 28 or dies prior to day 28, 
hospital free days will be zero. Patients transferred to another hospital or other health 
care facility will be followed to day 28 to assess this endpoint.  
7. Duration of ICU stay in survivors and non-survivors: The total number of days 
spent in the ICU until hospital discharge or death during the first 28 days. If a patient is 
discharged alive from the study hospital we assume they are no longer in the ICU.  
8. Initiation of assisted ventilation to day 28:  Any patient who received assisted 
ventilation in the first 28 days meets this endpoint. If a patient leaves the hospital 
without initiation of assisted ventilation, we assume they never started.  
9. Initiation of new renal replacement therapy to day 28.  
o Patients who receive (new) renal replacement therapy through day 28 will 
meet this endpoint.  
10. Change in organ-specific Sepsis-related Organ Failure Assessment (SOFA) 
scores between enrollment and day 7: We will calculate the SOFA score upon 


ASTER SAP  
 
pg. 9 
 
enrollment and at day 7 using clinically available data. If a value is not available at 
baseline, it will be assumed to be normal. At the day 7 assessment, if a value is 
missing then we will carry forward the closest previously known value. The GCS 
component of the SOFA score will be omitted.  (See Appendix B)  
11. 90-day hospital mortality (any hospital)  
12. Development of ARDS within 7 days: We will determine the presence and severity of 
ARDS for each day of assisted ventilation to day 7. For participants on assisted  
ventilation with P/F <300 or imputed P/F <300, FiO2 ≥40%, and PEEP ≥5 cm H2O, 
determine if hypoxemia is valid, acute, and not fully explained by CHF or fluid overload. 
If yes, local investigators will review the first CXR (or CT) performed on each ventilated 
day with valid P/F or imputed P/F <300 (to day 7). If no chest imaging studies are 
present that day, site investigators may review available imaging one day before or 
after to determine if ARDS imaging criteria are met. ARDS imaging criteria are met if 
the images are consistent with ARDS (bilateral opacities not fully explained by 
effusions, lobar/lung collapse, or nodules). If equivocal, the reviewing investigator will 
adjudicate with additional investigators.  
13. Change in serum creatinine from enrollment to discharge, death, initiation of dialysis, or 
day 28, whichever occurs first.  
14. Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline (defined as most recent outpatient creatinine from 24 
hours to 365 days prior to admission if available: if not available, lowest in hospital 
creatinine prior to randomization), need for renal replacement therapy, or death  
15. Change in Radiographic Assessment of Lung Edema (RALE) score from enrollment to 
study day 3 in patients who are receiving assisted ventilation or high flow nasal oxygen 
at the time of study randomization  
16. The safety of intravenous Acetaminophen compared to placebo as measured by daily 
serum AST, ALT, blood pressure following study drug administration (both arms) and 
incidence of reported adverse events (both arms)  
17. Change in plasma IL-6, plasma IL-8, plasma angiopoietin-2, plasma cell-free 
hemoglobin, syndecan-1, and plasma cell-free DNA from enrollment to days 2 and 3  
18. 90 day all-cause mortality: We will contact patients at day 90 to ascertain their 
survival status. This will be done by telephone contact with the patient or family 
members as well as a review of medical records and publicly available data sources. 
We will use the national death index as a final check for patients with whom we are 
unable to confirm their vital status through other means.  
 
2.3 Safety Endpoints  
AST and ALT will be measured on days 2, 3, 4, 5, and 7 after study enrollment. We will also 
collect data daily on potential adverse reactions to Acetaminophen, such as rash and 
hypersensitivity.  If either are present and thought to be due to the study drug, the study drug 
will be stopped.  


ASTER SAP  
 
pg. 10 
 
2.4 Predictive Biomarker Analysis  
We will evaluate the predictive value of baseline plasma CFH (Cell-free Hemoglobin) for 
identifying who have an improvement in the primary endpoint, the three components of the 
primary endpoint, 28-day mortality or improvement in renal function as assessed by MAKE28. 
2.5 Statistical Analysis Plan  
The principal analyses will be intention-to-treat (based upon randomization assignment). A 
comprehensive statistical analysis plan will be written prior to the un-blinding of the primary 
and secondary efficacy results.  
2.6 Interim Analysis  
The DSMB will have the ability to analyze all safety data, including AST/ALT, at any time 
during the study and stop the study indefinitely for safety reasons. The safety report will 
include a tabulation of all baseline demographics and physiologic variables as well as on-study 
variables, including SOFA scores, and all adverse events by randomized groups. For 
monitoring safety, data on 28-day mortality will be included with 95% confidence intervals for 
any differences between placebo and the acetaminophen group. In addition, a formal safety 
analysis will be done after 225 patients are accrued. This report will include an analysis of 
acetaminophen levels.  Trough levels greater than 20 mcg/mL will be evaluated for any 
relationship to AST/ALT levels.  Investigators will recommend protocol modification as needed 
to the DSMB as part of the safety review.   
Based on an interim analysis of the primary efficacy variable, this trial may stop for efficacy or 
futility of APAP relative to placebo. The stopping rules are based on two stage Haybittle-Peto 
stopping boundaries [1, 2] with Z=3.4 for early efficacy stopping. We plan one interim look at 
approximately one half of total enrollment and a final look at full enrollment.   
Table 1 below presents the stopping boundaries as a function of the mean difference in the 
primary efficacy variable favoring active treatment and the one-sided p-value favoring active 
treatment. Tables 2 and 3 present the probability of stopping at stage one under the null and 
alternative hypotheses respectively. As can be seen in Table 2 the probability of stopping early 
for futility under the null is 10%.  Table 3 shows that the probability of stopping early for 
efficacy under the alternative is also 10%.  
Table 1. Stopping Boundaries  
n 
Futility Bound  
Mean Difference 
Efficacy Bound  
Mean Difference 
Futility Bound  
P-value 
Efficacy Bound  
 P-value 
~218 
-1.51088  
4.01088 
0.89986 
0.0003369 
436 
1.63584 
1.63584 
0.02493 
0.02493 
 
Table 2. Stopping probabilities given the null hypothesis 
Stage  
Futility  
Efficacy  
1  
0.10014  
0.00034  
 
Table 3. Stopping probabilities given the alternative hypothesis 


ASTER SAP  
 
pg. 11 
 
Stage  
Futility  
Efficacy  
1  
0.00034  
0.10014  
  
 
3. Sub-Group by Treatment Interaction 
We will do a sub-group by treatment interaction analysis comparing outcomes in acute COVID-
19, defined as a positive diagnostic test for SARS CoV-2 in the prior three weeks and 
pneumonia as primary site of infection, and COVID-19 negative patients for the primary and 
secondary efficacy endpoints. The analysis will include a tabulation of the treatment effect in 
COVID-19 positive patients, the treatment effect in COVID-19 negative patients, and the 
difference between the treatment effect in these two sub-groups. We will also apply this 
interaction analysis separately to three demographic factors, gender (male versus female), 
race (White versus Non-White), ethnicity (Hispanic versus Non-Hispanic) and one additional 
pre-randomization factors, received acetaminophen (yes versus no). 
4. Statistical Methods 
Overview 
Unless otherwise specified all analyses are intention to treat, comparing the two treatment 
groups of those randomized to receive Acetaminophen and those randomized to receive 
placebo.  
Unless otherwise specified statistical significance will be indicated by two-sided p-values less 
than or equal to 0.05 with no adjustment made for multiple comparisons.  
Analysis is based on all available data with no imputation of missing data, except as needed to 
calculate selected derived variables, such as SOFA variables.  
The specified use of a Chi-square test may be superseded by use of Fisher’s exact test given 
sparse data. 
Primary Efficacy Outcomes 
Continuous primary efficacy outcomes will be compared between treatment groups using a t-
test. 
Binary primary efficacy outcomes will be compared between treatment groups using a Chi-
square test. 
Secondary Efficacy Outcomes 
Continuous secondary efficacy outcomes will be compared between treatment groups using a t-
test. 
Binary secondary efficacy outcomes will be compared between treatment groups using a Chi-
square test. 
 


ASTER SAP  
 
pg. 12 
 
Subgroup by Treatment Interactions for the Primary and Secondary Efficacy Outcomes 
Subgroup by treatment interactions for continuous efficacy outcomes will be assessed by an 
analysis of variance model including a treatment effect, a subgroup effect, and a subgroup by 
treatment interaction. Contrasts of the model coefficients will be used to estimate the treatment 
effect in each level of the subgroup variable.  
Subgroup by treatment interactions for binary efficacy outcomes will be assessed by a 
generalized linear model with a binary distribution function and identity link function. The model 
will include a treatment effect, a subgroup effect, and a subgroup by treatment interaction. 
Contrasts of the model coefficients will be used to estimate the treatment effect in each level of 
the subgroup variable.  
With 5 binary subgroup variables and N primary and secondary efficacy outcomes there will be 
a total of 5xN interaction p-values and 10*N p-values for the treatment effect within each 
subgroup level and over all p-values a total of 15*N. A Bonferroni multiple comparison 
adjustment over all 15*N p-values is extremely conservative and would probably eliminate 
detection of any potentially important findings. We therefore choose to designate this, a prior, an 
exploratory hypothesis generating analysis consistent with the early development phase of a 
new intervention, so that any findings would need future confirmation. Thus, we will impose no 
multiplicity correction and report nominal p-values. 
However, acknowledging the implications of multiplicity is also important to help interpret the 
trial results. Interpretation would depend on the research question of the reader. If the reader is 
interested in any interaction in one specific outcome, then there are 5 tests to adjust for. If the 
reader is interested in an interaction for any outcome for one subgroup of concern, then there 
are N tests to adjust for. If the reader is interested in any interaction for any outcome and 
subgroup, then there are 5*N tests to adjust for. Readers could use the appropriate Bonferroni 
correction to control the type 1 error rate for their specific research question.  
We will, therefore, present all the results with nominal p-values but also provide explanatory text 
and Bonferroni critical values consistent with various ways of sub-setting the results to specific 
research questions. 
Safety Outcomes 
Adverse events will be categorized by MedDRA system organ class and severity. For each 
category the proportion of patients having one or mor events will be compared between 
treatment groups with Fisher’s exact test. 
Acetaminophen trough levels in the first 100 evaluable patients, will be compared between 
treatment groups. A patient is “evaluable” if the patient received at least five doses of study drug 
prior to the day two sample draw, and the day two sample was drawn within 12 hours of 
administration of the previous dose. 
Trough APAP levels will be presented graphically with APAP levels displayed on the vertical 
axis and elapsed time between starting time of previous dose and trough level sample draw 
displayed on the horizontal axis. This plot will include superimposed established contours 
separating safe from unsafe levels [3]. One figure will be presented for patients on active 
treatment and one for patients on placebo treatment. 


ASTER SAP  
 
pg. 13 
 
New AST or ALT Elevation: defined as post randomization AST or ALT ≥ 10 x the ULN in 
patients who did not meet this criterion prior to randomization, will be compared between 
treatment groups using a Chi-square test. 
New Hy’s Law: defined as post randomization AST or ALT ≥ 3 x the ULN with bilirubin ≥ 2 x the 
ULN on the same calendar day in patients who did not meet this criterion prior to 
randomization[4], will be compared between treatment groups using a Chi-square test. 
Administration of fluid bolus within 120 minutes of study drug infusion: The number of patients 
given one or more fluid boluses will be compared between treatment groups using a Chi-square 
test. In those receiving one or more fluid boluses, the number of study drug doses followed by a 
fluid bolus in each patient will be compared between treatment groups using the CMH mean 
score test. The volume of fluid given in the largest fluid bolus in each patient will be compared 
between treatment groups using a T-test. 
New use of vasopressor within 120 minutes of study drug infusion: The number of patients 
given any new vasopressor will be compared between treatment groups using a Chi-square 
test. The number of new vasopressors given in each patient will be compared between 
treatment groups using the CMH mean score test. The maximum administration rate of any 
new vasopressor in each patient (in norepinephrine equivalents) will be compared between 
treatment groups using a T-test. 
Increased dose of vasopressor within 120 minutes of study drug infusion: The number of 
patients given one or more increased doses of vasopressors will be compared between 
treatment groups using a Chi-square test. The number of study drug doses followed by a 
vasopressor rate increase in each patient will be compared between treatment groups using the 
CMH mean score test. The maximum vasopressor increase in each patient (in norepinephrine 
equivalents) will be compared between treatment groups using T-test. 
Predictive Biomarker Analysis 
We will evaluate the predictive value of baseline plasma CFH for identifying patients who have 
an improvement in the primary endpoint, the three components of the primary endpoint, 28-day 
mortality or improvement in renal function as assessed by MAKE28.   
We will perform two analyses, one treating baseline plasma cell-free Hgb both as a continuous 
variable and one treating it as a binary indicator for plasma cell-free Hgb greater than 10 mg/dl 
in the analysis. The lower limit of detection is 0.00625 mg/dl.  
Baseline plasma cell-free Hgb by treatment interactions for continuous efficacy outcomes 
(primary endpoint, the three components of the primary endpoint) will be assessed by an 
analysis of variance model including a treatment effect, a Hgb effect, and a Hgb by treatment 
interaction.  
Baseline plasma cell-free Hgb by treatment interactions for binary efficacy outcomes (28-day 
hospital mortality, improvement in renal function as assessed by MAKE28) will be assessed by 
a generalized linear model with a binary distribution function and logit link function. The model 
will include a treatment effect, a Hgb effect, and a Hgb by treatment interaction.  
 


ASTER SAP  
 
pg. 14 
 
References 
[1] Martin, G.S., et al., The epidemiology of sepsis in the United States from 1979 through 2000. 
N Engl J Med, 2003. 348: p. 1546-1554.  
[2] Haybittle, J.L., Repeated assessment of results in clinical trials of cancer treatment. Br J 
Radiol, 1971. 44(526): p. 793-7.  
[3] Rumack, B. H., & Matthew, H. (1975). Acetaminophen poisoning and toxicity. Pediatrics, 55(6), 
871–876. 
[4] Guidance for Industry, Drug-Induced Liver Injury: Premarketing Clinical Evaluation: U.S. 
Department of Health and Human Services, Food and Drug Administration, Center for Drug 
Evaluation and Research (CDER), Center for Biologics Evaluation and Research (CBER), July 
2009, Drug Safety 
 
 


ASTER SAP  
 
pg. 15 
 
Table of Variables 
Variable 
Category 
Scale 
Method 
Notes 
Alive and free of organ 
support to day 28. 
Primary efficacy 
outcome 
Continuous 
T-test 
 
28-day day all-cause all-
location mortality  
Primary efficacy 
outcome component 
Binary 
Chi-square 
test 
 
Days free of assisted 
ventilation to day 28 in 
the overall cohort and 
survivors  
Primary efficacy 
outcome component 
Continuous 
T-test 
 
Days free of new RRT to 
day 28 in the overall 
cohort and survivors  
Primary efficacy 
outcome component 
Continuous 
T-test 
 
Days free of 
vasopressors to day 28 
in the overall cohort and 
survivors  
Primary efficacy 
outcome component 
Continuous 
T-test 
 
28-day ventilator-free 
days   
Secondary efficacy 
outcome 
Continuous 
T-test 
 
28-day vasopressor-free 
days   
Secondary efficacy 
outcome 
Continuous 
T-test 
 
28-day new renal 
replacement-free days  
Secondary efficacy 
outcome 
Continuous 
T-test 
 
28-day hospital mortality   
Secondary efficacy 
outcome 
Binary 
Chi-square 
test 
 
28-day ICU free-days  
Secondary efficacy 
outcome 
Continuous 
T-test 
 
28-day hospital-free 
days to discharge home  
Secondary efficacy 
outcome 
Continuous 
T-test 
 
28-day duration of ICU 
stay in survivors and 
non-survivors   
 
Secondary efficacy 
outcome 
Continuous 
T-test 
 
Initiation of assisted 
ventilation to day 28  
 
Secondary efficacy 
outcome 
Binary 
Chi-square 
test 
 
Initiation of renal 
replacement therapy to 
day 28  
 
Secondary efficacy 
outcome 
Binary 
Chi square 
test 
 
Change in Sequential 
Organ Failure 
Assessment (SOFA) 
scores between 
enrollment and study 
day 7  
 
Secondary efficacy 
outcome 
Continuous 
T-test 
 
90-day hospital mortality  
Secondary efficacy 
outcome 
Binary 
Chi-square 
test 
 
Development of ARDS 
within 7 days of 
randomization   
 
Secondary efficacy 
outcome 
Binary 
Chi-square 
test 
 
Change in serum 
creatinine from 
Secondary efficacy 
outcome 
Continuous 
T-test 
 


ASTER SAP  
 
pg. 16 
 
enrollment to discharge, 
death, initiation of 
dialysis or 28 days, 
whichever occurs first   
Major Adverse Kidney 
Events at 28 days 
(MAKE28):  
Secondary efficacy 
outcome 
Binary 
Chi-square 
test 
 
Change in the 
Radiographic 
Assessment of Lung 
Edema (RALE) score 
from enrollment to study 
day 3 in patients who 
are receiving assisted 
ventilation or high flow 
nasal oxygen at 
baseline.  
Secondary efficacy 
outcome 
Continuous 
T-test 
 
Change in plasma IL-6, 
plasma IL-8, plasma 
angiopoietin-2, plasma 
cell-free hemoglobin, 
syndecan-1, and plasma 
cell-free DNA from 
enrollment to days 2 and 
3  
Secondary efficacy 
outcome 
Continuous 
T-test 
 
90-day all-cause all-
location mortality   
Secondary efficacy 
outcome 
Binary 
Chi-square 
test 
 
Daily serum AST, ALT, 
blood pressure following 
study drug 
administration  
Safety outcome 
Continuous 
T-test 
 
Hypersensitivity or rash  
Safety outcome 
Binary 
Chi-square 
test 
 
AST and ALT measured 
on study days 0, 2-5, and 
7  
Safety outcome 
Continuous 
T-test 
Analyze by day 
Acetaminophen trough 
level  
Safety outcome 
Continuous 
 
 
Administration of fluid 
bolus within 120 minutes 
of study drug infusion 
Safety outcome 
 
 
This outcome is 
assessed with 
three questions 
on the CRF. 
New use of vasopressor 
within 120 minutes of 
study drug infusion 
 
Safety outcome 
 
 
This outcome is 
assessed with 
three questions 
on the CRF. 
Increased dose of 
vasopressor within 120 
minutes of study drug 
infusion 
Safety outcome 
 
 
This outcome is 
assessed with 
three questions 
on the CRF. 
Incidence of reported 
adverse events 
Safety outcome 
Count 
Poisson 
regression 
 
Predictive value of 
baseline plasma CFH 
by treatment  
Predictive Biomarker 
Analysis 
Binary/Continuous 
ANOVA and 
GLM 
 
Note: The Chi-square test may be replaced by Fisher’s exact test if data sparse. 
 

ICF_007.pdf: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER)  
Study Title: Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
 NCT04291508 
Document date: 7JUL2022 


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
1
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
Part 2 of 2: STUDY SITE INFORMATION 
Site Name:  
<< insert name of organization>> 
Site Principal Investigator:  
<< insert name & credentials of responsible PI >> 
Site Principal Investigator Contact:  
<<insert 10-digit phone for PI>> 
Mailing Address:  
<< insert PI mailing address>> 
Site Study Coordinator (Optional): 
<<Name & credentials of alternate POC>> 
Site Study Coordinator Contact: 
<<insert 10-digit phone for alternate POC>> 
This part of the consent form includes information about the site that is asking you to participate in this study 
and is specific to participation at your site only. Before making your decision, both the site-specific information 
and the general study information should be reviewed with you.  Your medical record will contain a note 
saying you are in a research study and may contain some research information about you.  Anyone you 
authorize to receive your medical record will also get this information. 
Costs to you if you take part in this study: 
  
Payment in case you are injured because of this research study: 
 
Who to call for any questions or in case you are injured: 
For additional information about giving consent or your rights as a person in this study, to discuss problems, 
concerns, and questions, or to offer input, please feel free to call the Vanderbilt University Medical Center 
Institutional Review Board Office at (615) 322-2918 or toll free at (866) 224-8273. 
Additional information about your local site: 
Confidentiality   
The research study Coordinating Center at Massachusetts General Hospital (MGH) in Boston, MA, or the other 
hospitals involved in this study may share your study information with doctors or researchers using it for other 
research projects not listed in this form, without any recipients knowing it is related to you specifically. It is 
possible that other researchers involved in this study may contact you in the future regarding potential 
participation in other studies and/or to see how you are doing. Other hospitals involved in this study, MGH, and 
their staff will comply with all laws regarding the privacy of such information. There are no plans to pay you for 
the use or transfer of this de-identified information. Your personal identifiable information will not be shared, 
unless required by government agencies that oversee or fund research.  


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
2
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
Certificate of Confidentiality  
We have obtained a Certificate of Confidentiality from the National Institutes of Health (NIH). This protects us 
and you from being forced to release study information as part of a court, legislative, administrative or other 
proceeding. 
There are times when the Certificate cannot be used. For example, we cannot refuse to give information to 
government agencies that oversee or fund research, such as the NIH, Department of Health and Human Services 
(DHHS) or Food and Drug Administration (FDA).  
The Certificate also does not stop us from giving information to government agencies, law enforcement 
personnel, or others if we suspect you or someone else is in danger.  We can release some study information, 
such as laboratory test results, if you wish us to do so and you give permission in writing. The Certificate does 
not keep you from giving out information about yourself and your treatment in this study. If you would like to 
read the Certificate or you have any questions, please ask the study doctor or study staff.  
Authorization to Use/Disclose Protected Health Information 
 
 
 


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
3
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
If you decide not to take part in this research study, it will not affect your treatment, payment or enrollment 
in any health plans or affect your ability to get benefits. 
You will get a copy of this form after it is signed. 


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
4
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
 
 


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
5
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
CONSENT FOR GENETIC TESTING ON STORED SPECIMENS 
The purpose of this part of the study is to store samples of your blood. We are looking at genetic factors (such 
as DNA or RNA) that may cause or relate to severe illness or other diseases.  
Description of the Procedures: An extra 1 teaspoon of blood will be drawn and stored for genetic testing 
purposes. Whenever possible, the blood will be taken with other laboratory test samples to avoid an extra 
needle stick. Your blood sample will be processed and may be tested and shared for research genetic testing. 
The sample will be sent to a repository at the NIH. The NIH stores and distributes blood samples and associated 
data from people with many conditions. The purpose of sending your blood samples to the NIH is to make 
samples available for future research by investigators not involved in this study. Researchers who use samples 
from the NIH must request and receive approval to do so from the NIH and from research oversight boards at 
their institutions. 
Confidentiality of Your Blood Samples: We will freeze your samples and store them for years for future studies. 
We will protect your privacy and give all samples coded study numbers. Only your hospital study team will know 
your coded study number identity, which will be kept secure. The stored blood samples will not contain your 
name or identifying information.  
How Long Will the Samples Be Stored: The samples will be stored for an unknown period time (usually years). 
The samples may be thrown away when they are no longer needed. The results of tests run on your samples are 
not in your normal medical record and neither you nor your doctor will be told of the results. No one else, 
including relatives, doctors, or insurance companies can get the stored samples or results. Your samples will be 
used only for research and will not be sold or used directly to produce commercial products. 
Risks: One possible risk might be the release of your name, which could link you to the stored samples and/or 
the results of the tests run on your samples. This could cause problems with insurance or getting a job. You are 
currently protected from genetic discrimination by employers or insurance companies through the Genetic 
Information Nondiscrimination Act (GINA 2008). To protect you from this risk we will keep the link between the 
samples and your personal ID as secure as possible. This link will only be kept by the local study team. The NIH 
repository and future researchers will not have access to any of your personal information so they will not know 
who you are or be able to contact you.  
Benefits: You will not receive any direct benefit from your samples. Information obtained from the tests may 
provide useful information to help other patients about the causes, risks, and prevention of severe illness and 
other diseases.  
Voluntary Participation/Right to Withdraw Your Permission for Genetic Testing: Your decision to join this study 
is completely up to you (voluntary). Your decision will not change the quality of the care you receive. You are still 
eligible to join the study described in the other consent form even if you do not want your blood samples stored 
for genetic testing. At any time, you may ask to have your samples destroyed. You should contact Dr. [PI Name] 
in writing to have your samples destroyed and no longer used for research.  We will not be able to destroy 


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
6
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
research data that has already been gathered using your samples.  Also, if your identity was removed from the 
samples, we will not be able to locate and destroy them.  
Costs or compensation of study: There will be no costs or compensation to you. 
Commercialization: Your samples may be used to make new products or tests.  These may have value and may 
be developed and owned by the study staff, [Name of Institution], and/or others.  If this happens, there are no 
plans to provide money to you. 
 
 
 


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
STUDY SITE INFORMATION  
Study Title: 
Acetaminophen in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date: 
 [cIRB Submission Date]  
  
  
7
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
CONSENT FOR CONTINUED RESEARCH PARTICIPATION 
You have been taking part in the research study: Acetaminophen in Sepsis: Targeted Therapy to Enhance 
Recovery (ASTER).  Consent for your participation was obtained from your legal representative because you 
were unable to provide consent at that time.  We are now asking for you to consent to continue being in the 
study.  Your continued participation is entirely voluntary.  If you decide not to continue in this study, it will not 
affect your relationship with your doctor or with [Hospital Name] and will not result in any penalty or loss of 
benefits to which you are otherwise entitled. 
STATEMENT OF VOLUNTARY CONSENT 
I have read this form and the attached consent or have had them read to me.  I have been told what to expect if 
I take part in this study, including risks and possible benefits.  I have had a chance to ask questions and have had 
them answered to my satisfaction.  I have been told that the people listed in this form will answer any questions 
that I have in the future.  By signing below, I am volunteering to continue to be in this research study. 
 

Prot_001.pdf: 
 
 
 
Acetaminophen in Sepsis: Targeted Therapy to 
Enhance Recovery (ASTER) 
 
 
ASTER: A PETAL Network Multi-Center Phase 2b Randomized Double-Blinded Placebo-
Controlled Trial originally designed to evaluate Two Different Pharmacologic Therapies 
(Intravenous Vitamin C or Intravenous Acetaminophen).  The Vitamin C arm was stopped on 
June 15, 2022.  
VERSION: 4.5 April 25, 2023 
PROTOCOL CO-CHAIRS  
  
Lorraine Ware, MD and Michael Matthay, MD (Co-Chairs Acetaminophen)  
Berry Fowler, MD and D. Clark Files, MD (Co-Chairs Vit C)  
 
PROTOCOL COMMITTEE  
  
 
               
 ADVISORS TO PROTOCOL  
  
Neil Aggarwal, MD 
Rebecca Baron, MD   
Antonello Punturieri, MD, PhD 
Karen Bienstock, PA-C 
Samuel Brown, MD    
  
Steven Chang MD, PhD  
Ivor Douglas, MD  
  
                             Roy G Brower, MD  
Abhijit Duggal, MD 
Scott Fields, PharmD  
Adit A. Ginde, MD  
Andrew Goodwin, MD 
Duncan Hite, MD  
Estelle Harris, MD  
Greg Hendey, MD  
Kathryn Hibbert, MD 
Terri Hough, MD   
Akram Khan, MD  
Kathleen Liu, MD, PhD  
Chadwick Miller, MD  
Ari Moskowitz, MD 
Richard Riker, MD 
Bryce RH Robinson, MD  
Wesley H. Self, MD  
B. Taylor Thompson, MD 
Mark Tidswell, MD  
Donald M. Yealy MD  
STUDY DRUG  
 Acetaminophen 
  
 
ASTER
TRIAL
APAP


ASTER Protocol Version 4.5  
April 25, 2023 
 
 
 
 
 
 
 
 
 
 
 Pg.  2     
  
  
  
Contents 
1. Abbreviations and Definitions .................................................................................................................. 5 
1.1 Abbreviations ......................................................................................................................................... 8 
1.2 Definitions ............................................................................................................................................. 10 
2. Trial Summary ......................................................................................................................................... 12 
2.1 Title ....................................................................................................................................................... 12 
2.2 Objective ............................................................................................................................................... 12 
2.3 Hypotheses ........................................................................................................................................... 12 
2.4 Study Design ......................................................................................................................................... 12 
2.4.1 Treatment Arms .................................................................................................................... 12 
2.4.2 Sample Size/Statistical Considerations: ................................................................................ 13 
2.5 Inclusion Criteria ................................................................................................................................... 14 
2.6 Exclusion Criteria .................................................................................................................................. 14 
2.7 Primary Efficacy Variable ...................................................................................................................... 15 
2.8 Secondary Clinical Efficacy Variables .................................................................................................... 15 
2.9 Safety Variables .................................................................................................................................... 16 
2.10 Biological Endpoints ............................................................................................................................ 16 
3. Trial Description...................................................................................................................................... 16 
3.1 Background ........................................................................................................................................... 16 
3.1.1 Background-Sepsis and sepsis-related organ dysfunction ................................................... 16 
3.1.2       Cell-free hemoglobin (CFH) and organ dysfunction in sepsis ......................................... 17 
3.2     Acetaminophen—Preclinical and Clinical Data ................................................................................. 18 
3.3 Summary ............................................................................................................................................... 20 
4. Trial Design ............................................................................................................................................. 21 
4.1 Study Description ................................................................................................................................. 21 
4.2 Randomization ...................................................................................................................................... 22 
4.3 Primary Efficacy Variable ...................................................................................................................... 22 
4.4 Secondary Clinical Efficacy Variables .................................................................................................... 23 
4.5 Safety Variables .................................................................................................................................... 24 
4.6 Biological Endpoints .............................................................................................................................. 24 
5. Treatments Arms and Study Drug Administration ................................................................................. 24 
5.1 Acetaminophen-Active Arm ................................................................................................................. 24 


ASTER Protocol Version 4.5  
April 25, 2023 
 
 
 
 
 
 
 
 
 
 
 Pg.  3     
5.2 Acetaminophen-Placebo Arm ............................................................................................................... 25 
5.3 Drug Interruptions ................................................................................................................................ 26 
6. Study Population and Enrollment ........................................................................................................... 26 
6.1 Participant Enrollment .......................................................................................................................... 26 
6.2 Inclusion Criteria ................................................................................................................................... 26 
6.3 Exclusion Criteria .................................................................................................................................. 27 
6.4     Reasons for each of the exclusion criteria ........................................................................................ 27 
6.5 Co-enrollment ....................................................................................................................................... 28 
6.6 Informed Consent ................................................................................................................................. 28 
6.7 Excluded Medications ........................................................................................................................... 29 
6.8 On-study Fever Management Recommendations ............................................................................... 29 
6.9 Concomitant Medications .................................................................................................................... 29 
6.10 Common Strategies for All Treatment Arms ...................................................................................... 29 
6.10.1 Ventilator Procedures ......................................................................................................... 29 
6.10.2   On Study Fluid Management ............................................................................................ 30 
6.11 Criteria for Discontinuation or Withdrawal ........................................................................................ 30 
7.0 Study Plan ............................................................................................................................................. 30 
7.1 Baseline Assessments ........................................................................................................................... 30 
7.2. Assessments During Protocol Phase (through study day 7) ................................................................ 31 
7.3. Other Assessments and Data Collection .............................................................................................. 32 
7.4. Assessments after Hospitalization ....................................................................................................... 32 
7.5. 90-day All-Cause Mortality .................................................................................................................. 32 
7.6. Specimen Collection ............................................................................................................................ 32 
8. Statistical Methods ................................................................................................................................. 33 
8.1 Primary Endpoint .................................................................................................................................. 33 
8.2   Sample Size ......................................................................................................................................... 34 
8.3 Secondary endpoints ............................................................................................................................ 34 
8.4 Safety Endpoints ................................................................................................................................... 36 
8.5 Predictive Biomarker  Analysis ............................................................................................................. 36 
8.6 Statistical Analysis Plan ......................................................................................................................... 36 
8.6.1 Sub-Group by Treatment Interaction ................................................................................... 36 
8.7 Interim Analysis .................................................................................................................................... 36 
9.0 Risk Assessment .................................................................................................................................... 37 


ASTER Protocol Version 4.5  
April 25, 2023 
 
 
 
 
 
 
 
 
 
 
 Pg.  4     
9.1 Potential Risks to Subjects .................................................................................................................... 37 
9.2  Risk of Death ........................................................................................................................................ 38 
9.3  Minimization of Risk ............................................................................................................................ 38 
9.4 Potential Benefits ................................................................................................................................. 38 
9.5 Risks in Relation to Anticipated Benefits .............................................................................................. 38 
10. Human Subjects .................................................................................................................................... 39 
10.1 Selection of Subjects ........................................................................................................................... 39 
10.2   Justification for Including Vulnerable Subjects ................................................................................. 39 
10.3 Informed Consent ............................................................................................................................... 39 
10.3.1 Process of Obtaining Informed Consent ............................................................................. 39 
10.4   Continuing Consent ........................................................................................................................... 42 
10.5 Withdrawal of Consent ....................................................................................................................... 42 
10.6    Identification of Legally Authorized Representatives ...................................................................... 42 
10.7    Justification of Surrogate Consent ................................................................................................... 43 
10.8     Additional Safeguards for Vulnerable Subjects ............................................................................... 43 
10.9     Confidentiality ................................................................................................................................. 43 
11. Adverse Events ..................................................................................................................................... 44 
11.1 Safety Monitoring ............................................................................................................................... 44 
11.2 Serious Adverse Events ....................................................................................................................... 44 
Appendices ................................................................................................................................................. 46 
A. Schedule of Events ................................................................................................................................. 46 
B. Sequential (Sepsis-related) Organ Failure Assessment (SOFA) Score .................................................... 47 
C. Ventilator Management for Patients with ARDS .................................................................................... 48 
D. On-Study Fluid Management for Patients with ARDS ............................................................................ 50 
E. Adverse Event Reporting and Unanticipated Events .............................................................................. 51 
E.1. Unanticipated Problems (UP) ................................................................................................. 51 
E.2. Determining Relationship of Adverse Events to Procedures .................................................. 51 
E.3. Clinical Outcomes that may be Exempt from Adverse Event Reporting ................................ 52 
F. Alcohol Use Screen Audit C ..................................................................................................................... 53 
G. Fever Management Guidelines .............................................................................................................. 54 
References .................................................................................................................................................. 55 
 
  


  
ASTER Protocol Version 4.4 
July 7, 2022 
 
 
 
 
 
 
 
 
 
 
Pg.  5     
Revisions to the Protocol 
Protocol Version 4.5 
Date: April 25, 2023  
Substantive protocol changes in Version 4.5:  
1. Updates to the Protocol Cover Page were made to match changes in the protocol 
committee members’ list.    
 
Protocol Version 4.4 
Date: July 7, 2022  
On June 15, 2022, the study was paused due to publication of the Lessening Organ dysfunction 
with Vitamin C [1] trial demonstrating that intravenous Vitamin C at the dose used in this study 
was unlikely to benefit a similar population of participants as enrolled in ASTER and may be 
harmful [1]In addition, the sample size was revised based on a new estimate of the standard 
deviation of the primary outcome (organ support free days; OSFDs) to support the feasibility of 
completing the trial while retaining 85% power to detect the originally postulated 2.5 OSFD day 
difference with acetaminophen. Lastly, certain aspects of the protocol including exclusions that 
were relevant only to the vitamin C arm were removed. 
 
Substantive protocol changes in Version 4.4: 
1. Removal of all language related to Vitamin C from the following sections:  
• 
1. Abbreviations and Definitions 
• 
2.1 Title 
• 
2.2 Objective, 2.3 Hypotheses, 2.4 Study Design, 2.4.1 Treatment Arms, 2.4.2 
Sample Size/Statistical Considerations (sample size changed from 900 to 450 
and Acetaminophen/placebo arms changed from 300 to 225), 2.6 Exclusion 
Criteria (#7, 13, 14, and 21 removed), 2.7 Primary Efficacy Variable, 2.8 
Secondary Clinical Efficacy Variables, and 2.10 Safety Variables 
• 
3.1.2 Cell-free hemoglobin (CFH) and organ dysfunction in sepsis 
• 
Removal of section 3.2 Vitamin C: Preclinical and Clinical Data 
• 
3.4 Summary 
• 
4.1 Study Description, 4.2 Randomization, 4.3 Primary Efficacy Variable, 4.4 
Secondary Clinical Efficacy Variables, and 4.5 Safety Variables 
• 
5.3 Vitamin C Arm-Active and 5.4 Vitamin C-Placebo Arm sections removed and 
5.1 Acetaminophen-Active, 5.2 Acetaminophen-Placebo Arm sections revised to 
“Acetaminophen Arm” and “Placebo Arm” 
• 
6.3 Exclusion Criteria (#7, 13, 14, and 21 removed) and 6.4 Reasons for each of 
the exclusion criteria 


  
ASTER Protocol Version 4.4 
July 7, 2022 
 
 
 
 
 
 
 
 
 
 
Pg.  6     
• 
6.5 Co-enrollment 
• 
6.7 Excluded Medications and 6.8 On-study Fever Management 
Recommendations 
• 
Section 6.10 Blood Glucose Strategy for the Vitamin C-Active/Vitamin C-Placebo 
Arm removed 
• 
6.12 Criteria for Discontinuation or Withdrawal 
• 
7.5 90-day All-Cause Mortality 
• 
7.6 Sample Collection; Biological Endpoint Assays bullet 
• 
8.1 Primary Endpoint  
• 
8.2 Sample Size (stopping boundaries revised for sample size without Vitamin C 
arm and single placebo group) 
• 
8.3 Secondary endpoints (Renal calculi endpoint also removed) 
• 
8.5 Safety Endpoints 
• 
8.7 Interim Analysis (subject enrollment numbers changed from 450 to 218 in 
addition to removal of Vitamin C language, corresponding stopping boundaries 
updated, and paragraph after Table 3 removed) 
• 
9.0 Risk Assessment, 9.1 Potential Risks to Subjects, and 9.4 Potential Benefits 
• 
11.1.1 Safety Monitoring 
• 
Appendix A. Schedule of Events: “or vitamin C” removed from non-study 
administration row and Assessment of kidney stones removed from Outcome 
data section.  
• 
Appendix E. Adverse Event Reporting and Unanticipated Events 
 
2. Revision of exclusion criterion #15 (Protocol Sections 2.6 and 6.3: Exclusion Criteria):  
• 
Current language: Use of home oxygen for chronic cardiopulmonary disease 
• 
Revised language: Use of home oxygen > 3 liters/min nasal cannula for chronic 
cardiopulmonary disease  
 
Rationale for this change:  
 
Exclusion modification: 
• 
A modest requirement for home oxygen to treat cardiorespiratory disease could 
be reasonably described as chronic respiratory insufficiency, rather than chronic 
respiratory failure. Described this way, chronic respiratory insufficiency could be 
analogous to patients with chronic renal insufficiency manifesting as reduced 
GFR without a requirement for chronic dialysis. Patients with reduced GFR are 
eligible for ASTER, provided they meet other inclusion criteria.    
• 
Based on the most recent ASTER monthly report (SeeTable 2), 120 participants 
have been excluded because 
of the use of home oxygen 
for chronic cardiopulmonary 
disease, 86 as the sole 
reason for exclusion, and 34 


  
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as one of many reasons for exclusion. At actively screening sites, this is the 
number one reason to exclude a potential ASTER participant. If 86 additional 
participants had been included in the study as of May 2022, enrollment to-date 
would have been increased by 60-70%.  
• 
Interpretation of the study’s primary efficacy variable, number of organ failure 
free days to day 28, would not be affected by modifying this exclusion. To meet 
the respiratory system component of the primary efficacy variable, the participant 
must require assisted ventilation for at least 1 day. Assisted ventilation is 
currently defined in the ASTER trial as any level of invasive or noninvasive 
ventilator support including continuous positive airway pressure of >5 cmH20, 
bilevel positive pressure ventilation, or invasive mechanical ventilation. 
Noninvasive ventilation solely for sleep disordered breathing and nasal high flow 
oxygen therapy are not considered “assisted ventilation” for this study. 
• 
Though patients with a home oxygen requirement may be more susceptible to 
acute respiratory failure, we anticipate randomization would obviate any arm-
specific bias introduced by this increased susceptibility.  
• 
There is no biologic rationale for why a patient with underlying chronic respiratory 
insufficiency might not benefit from APAP  
• 
To aid the study team in interpreting participant eligibility, we will create a FAQ that 
helps clarify that if a participant meets inclusion criteria based on a 6L nasal cannula 
oxygen requirement, the infection (or presumed infection) should be the primary 
reason for an increase in oxygen needs from any baseline oxygen requirement.   
3. Correction to study drug administration in the Schedule of Events (SOE) table: 
Administration of study drug was missing from the Baseline (Day 0) timepoint in the 
table. The SOE table in Appendix A has been corrected to reflect study drug 
administration of the first dose on Day 0 (within 4 hours of randomization).  
 
Protocol Version 4.3 
Date: November 30, 2021 
Initial protocol 
 
 


  
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1. Abbreviations and Definitions   
1.1 Abbreviations   
ABG = Arterial Blood Gas  
APAP = Acetaminophen  
ALI = Acute Lung Injury  
ALT = Alanine Aminotransferase  
ARDS = Acute Respiratory Distress Syndrome  
AST = Aspartate Aminotransferase  
BIPAP = Bi-level Positive Airway Pressure  
BMI = Body mass index  
BP = Blood Pressure  
CCC = Clinical Coordinating Center  
CFH = Cell-free hemoglobin  
CPAP = Continuous Positive Airway Pressure  
CRF = Case Report Form  
Day 0 =  Day of Randomization  
DSMB = Data and Safety Monitoring Board  
ELISA = Enzyme-linked Immunosorbent Assay  
  
  
 
 
 
 
 
 
 
ED = Emergency Department  
F2-IsoP = F2-Isoprostanes  
FDA = Food and Drug Administration  
FiO2 = Fraction of Inspired Oxygen  
GCS = Glasgow Coma Scale  
HFNO = High Flow Nasal Oxygen  
HUVEC = Human Umbilical Vein  
Endothelial Cells  
ICF = Informed Consent Form  
ICU = Intensive Care Unit  
IMV = Intermittent Mechanical Ventilation  
IL-6 = Interleukin-6  
IL-8 = Interleukin-8  
IRB = Institutional Review Board  
IV = Intravenous  
LAR = Legally Authorized Representative  
LPS = Lipopolysaccharide  
MAP = Mean arterial blood pressure  
MBW = Measured Body Weight  
mL = Milliliter  
mmHg = Millimeter of Mercury  
NHLBI = National Heart Lung and Blood  
Institute  
NIV = Non-invasive ventilation   


  
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NSAID = Nonsteroidal anti-inflammatory drug  
PaO2 = Partial pressure of arterial oxygen  
PBW = Predicted Body Weight  
PEEP = Positive End-Expiratory Pressure   
Pplat = Plateau pressure  
PSV = Pressure Support Ventilation  
POC = Point of Care  
RALE = Radiographic Assessment of Lung Edema  
RCT = Randomized Controlled Trial  
RRT = Renal Replacement Therapy  
SAE = Serious Adverse Event  
SBP = Systolic Blood Pressure  
SBT = Spontaneous Breathing Trial  
SOFA = Sequential (Sepsis-related) Organ Failure 
Assessment   
SpO2 = Oxygen Saturation by Pulse  
Oximetry  
SUSAR = Suspected and Unexpected Serious 
Adverse Reaction  
UAB = Unassisted Breathing  
VFD = Ventilator-free Days  
 
  
  
  
  


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1.2 Definitions  
Adverse Event: Any untoward medical occurrence associated with the use of a drug or a study 
procedure, whether or not considered drug related.  
Adverse Reaction: Any adverse event caused by a study intervention. An adverse reaction is a 
subset of all suspected adverse reactions where there is a reason to conclude that the study 
intervention caused the event.   
Assisted ventilation: Any level of invasive or noninvasive ventilator support including 
continuous positive airway pressure of >5 cmH20, bilevel positive pressure ventilation, or 
invasive mechanical ventilation.  Noninvasive ventilation solely for sleep disordered breathing 
and nasal high flow oxygen therapy are not considered “assisted ventilation” for this study.  
Baseline: Prior to randomization  
Chronic Dialysis: Receipt of hemodialysis or peritoneal dialysis for renal failure prior to 
hospitalization.  This does not include new renal replacement therapy or hemofiltration initiated 
within 36 hours of randomization. 
Date of first Unassisted Breathing (UAB): Defined as the first day that the subject is on UAB 
from midnight to midnight. Example: if subject meets UAB at 1900 on 6/1/14, then the date of 
first UAB would be 6/2/14, as long as subject does not return to AB on 6/2/14.  
Day zero: Defined as the day of randomization (until 23:59 of the study day).  
Days Alive and Free of Organ Support: Days alive and free of assisted ventilation, 
vasopressors and new renal replacement therapy over 28 days  
Extubation: Removal of an orotracheal or nasotracheal tube, or unassisted breathing with a 
tracheostomy  
Fluid Bolus: 500 ml or more of isotonic crystalloid  
High Flow Nasal Oxygen: Oxygen delivered by nasal cannula at a rate of at least 30 liters/min, 
using a system capable of delivering up to 100% humidified and heated oxygen at flow rates up 
to 60 liters per minute.  
Home: Level of residence or health care facility where the patient was residing prior to hospital 
admission  
Invasive Mechanical Ventilation: Mechanical ventilation delivered through an endotracheal or 
tracheostomy tube.  
Legally Authorized Representative: Any individual, judicial, or other body authorized under 
applicable law to consent on behalf of a prospective patient to the patient’s participation in the 
clinical study  
Serious Adverse Event: Adverse events that are serious and have a reasonable possibility that 
the event was due to a study drug or procedure.  


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Study hospital: Defined as the hospital where the patient was randomized and enrolled.  
Study withdrawal: Defined as permanent withdrawal from study the before completion of study 
activities. This does not include those subjects who have completed the protocol procedures. If 
a patient or surrogate requests withdrawal from the study the investigator should seek explicit 
permission to continue data collection.  
Suspected Adverse Reaction: Any adverse event for which there is a reasonable possibility 
that the study procedures caused the adverse event. Reasonable possibility means there is 
evidence to suggest a causal relationship between the study procedures and the adverse event. 
A suspected adverse reaction implies a lesser degree of certainty about causality than adverse 
reaction.   
Suspected Unexpected Serious Adverse Reaction (SUSAR): An adverse reaction that is 
both unexpected (not consistent with risks outlined in the protocol or investigator brochure), 
serious, and meets the definition of a suspected adverse reaction.  
UAB (Unassisted Breathing):  Spontaneously breathing with face mask, nasal prong oxygen,  
or room air, T-tube breathing, tracheostomy collar (mask) breathing, or CPAP ≤ 5 without PSV 
or IMV assistance, nasal high flow oxygen, or the use of noninvasive ventilation solely for sleep-
disordered breathing.  Assisted breathing is any level of ventilatory support at pressures higher 
than the unassisted breathing thresholds.  
 
 


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2. Trial Summary  
2.1 Title  
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
ASTER is a platform originally designed to conduct two separate trials. In response to external 
trial results, enrollment in the Vitamin C arm of the trial was suspended on June 15, 2022.  The 
Acetaminophen (APAP) arm of the trial was resized and will continue as described in this 
amended protocol. 
2.2 Objective  
Objective: To carry out one multi-center phase 2b randomized double-blinded placebo-
controlled trial of one pharmacologic therapy. The trial will assess the efficacy of 
Acetaminophen (1 gram intravenously every 6 hours) in comparison to placebo for 120 hours in 
patients with sepsis who have evidence of either hemodynamic or respiratory organ failure. The 
placebo infusion (5% dextrose in water, D5W) will match the volume and storage temperatures 
of the acetaminophen (room temperature, dose adjusted for weight below 50 kg)  
2.3 Hypothesis  
Hypothesis: Acetaminophen (APAP) infusion will increase the days alive and free of organ 
support to day 28.  
2.4 Study Design  
Prospective multi-center phase 2b randomized placebo-controlled double-blinded interventional 
trial of intravenous Acetaminophen for patients with sepsis-induced hypotension or respiratory 
failure.   
1. Enrollment Period: Approximately 15 months  
2. Patient Population: Critically ill adults admitted or planned admission to the ICU with 
sepsis and either hemodynamic or respiratory organ failure.  A maximum of 450 patients 
will be enrolled  
3. Locations: Emergency Department and ICUs at hospitals participating in the NHLBI 
PETAL Network  
2.4.1 Treatment Arms 
Patients with sepsis and either hemodynamic or respiratory organ failure on enrollment will be 
randomized to one of two treatment arms in a 1:1 fashion.  The two  arms will be 
Acetaminophen and Placebo randomized 1:1.  


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Patients, nurses, research staff, and physicians will be blinded to the treatment assignment. The 
time of randomization will represent time zero.  Study drug will be started within 4 hours of 
randomization. 
 
2. Completion of study drug administration: Study drug administration will occur for a 
total of 120 hours, unless one of the following occurs:  
1. Discharged from the study hospital  
2. Discharge from the ICU  
3. Withdrawal from the study   
4. Death  
5. New AST or ALT elevation ≥ 10 times the upper limit of normal -  
2.4.2 Sample Size/Statistical Considerations:  
With randomization of 450 total patients (225 Acetaminophen 225 placebo)we will have 85% 
power to detect a difference between groups of 2.5 days in the primary composite outcome of 
1. Acetaminophen given intravenously at the dose of 1 gram (or 15 mg/kg if patient 
weighs < 50 kg) every six hours for 5 days (20 doses) OR 
2. Placebo (identical appearing 5% dextrose solution) infused every six hours for 5  days 
(20 doses)  
NOTE: The volume of placebo infusions will be lower in patients <50kg and will be 
matched to Acetaminophen.  
Treatment will 
continue for 120 
hours (20 doses), or 
discharge from the 
intensive care unit, 
study withdrawal, or 
death, whichever 
comes first. 
First study drug dose  
(APAP or placebo) will be 
considered “Dose 1” and 
will be administered within 
4 hours of 
randomization.   
Study drug will be infused 
over 30 minutes.  
 
Subsequent doses will be infused 
every six hours (+/- 1 hour) 
through 120 hours or 20 doses 
(whichever comes first).  If a dose 
cannot be administered within 3 
hours of the scheduled time, the 
dose should be skipped.   
Any dose administered outside of 
the +/- 1 hour window or any dose 
that is skipped will trigger a protocol 
deviation 
Duration 
Subsequent Dosing 
Timing of Dosing 


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days alive and free of organ support (dialysis, assisted ventilation, and vasopressors) to day 28. 
Patients will be analyzed on an intention-to-treat basis based on randomization assignment.  
 
2.5 Inclusion Criteria  
1. Age ≥ 18 years  
2. Sepsis defined as:  
a. Clinical evidence of a known or suspected infection and orders written to administer 
antibiotics  
AND  
b. Hypotension as defined by the need for any vasopressor (and at least 1 liter of fluid 
already administered intravenously for resuscitation) OR respiratory failure defined 
by mechanical ventilation, BIPAP or CPAP at any level, or greater than or equal to 6 
liters/minute of supplemental oxygen (criterion b must be present at the time of 
randomization)  
3. Admitted (or intent to admit) to a study site ICU within 36 hours of presentation to the ED 
or any acute care hospital  
2.6 Exclusion Criteria  
1. No consent/inability to obtain consent from the participant or a legally authorized 
representative  
2. Patient unable to be randomized within 36 hours of presentation to the ED or within 36 hours 
of presentation to any acute care hospital  
3. Diagnosis of cirrhosis by medical chart review  
4. Liver transplant recipient   
5. AST or ALT greater than five times the upper limit of normal   
6. Diagnosis of ongoing chronic alcohol use disorder/abuse by chart review; if medical record 
unclear, use Appendix F  
7. Hypersensitivity to Acetaminophen  
8. Patient, surrogate or physician not committed to full support (Exception: a patient will not be 
excluded if he/she would receive all supportive care except for attempts at resuscitation from 
cardiac arrest)  
9. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used 
only for sleep-disordered breathing  
10. Chronic dialysis 
11. Use of home oxygen > 3 liters/min nasal cannula for chronic cardiopulmonary disease  
12. Moribund patient not expected to survive 24 hours   
13. Underlying malignancy or other condition with estimated life expectancy of less than 1 month  


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14. Pregnant woman, woman of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or woman who is breast feeding  
15. Prisoner  
16. Treating team unwilling to enroll because of intended use of Acetaminophen  
2.7 Primary Efficacy Variable  
Primary efficacy variables will be evaluated comparing Acetaminophen vs. placebo The primary 
efficacy variable is days alive and free of organ support (dialysis, assisted ventilation, and 
vasopressors) to day 28.  
Participants will need to be free of all three components (assisted ventilation, vasopressors, new 
renal replacement therapy) to qualify for a day alive and free from organ failures. A patient could 
be free of all organ failures from day 10 through day 22 and then die on day 22 and would 
receive 12 days of credit for this primary outcome.  The components of this outcome (days free 
of assisted ventilation, vasopressors, and new renal replacement therapy in the overall cohort 
and in survivors, and 28-day hospital mortality) will also be reported as secondary outcomes 
(see below in 2.8).  
2.8 Secondary Clinical Efficacy Variables  
Secondary efficacy variables will be evaluated comparing Acetaminophen vs. placebo.  
1. 28-day ventilator-free days   
2. 28-day vasopressor-free days   
3. 28-day new renal replacement-free days  
4. 28-day hospital mortality   
5. 28-day ICU free-days  
6. 28-day hospital-free days to discharge home  
7. 28-day duration of ICU stay in survivors and non-survivors   
8. Initiation of assisted ventilation to day 28  
9. Initiation of renal replacement therapy to day 28  
10. Change in Sequential Organ Failure Assessment (SOFA) scores between enrollment 
and study day 7  
11. 90-day hospital mortality (includes death at any healthcare facility that is not the 
equivalent of home for a patient)  
12. Development of ARDS within 7 days of randomization   
13. Change in serum creatinine from enrollment to discharge, death, initiation of dialysis or 
28 days, whichever occurs first   
14. Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline, need for new renal replacement therapy, or death  


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15. Change in the Radiographic Assessment of Lung Edema (RALE) score from enrollment 
to  study day 3 in patients who are receiving assisted ventilation or high flow nasal 
oxygen at baseline.  
16. 90-day all-cause mortality   
2.9 Safety Variables  
The safety monitoring and analyses will be performed separately on the Acetaminophen and 
placebo arms (1 active to 1 placebo randomization ratio). The safety variables will include the 
following:  
1. Hypersensitivity or rash  
2. AST and ALT measured on study days 0, 2-5, and 7 (day 7 measurement can be ±1 day).  
3. The plasma levels of Acetaminophen will be obtained as a trough level just prior to 
administration of a study dose on study day 2 (subject must have received at least 5 doses 
of study drug before the day 2 samples are obtained) for all patients. After the first 100 
patients have been enrolled who have been randomized to the Acetaminophen arm, the 
trough levels will be reported to the DSMB to determine if they exceed the level of 20 
micrograms/mL, the potential level for hepatotoxicity.  
4. Administration of fluid bolus, new use of vasopressor, or increased dose of vasopressor 
within 120 minutes of study drug infusion.  
5. Incidence of reported adverse events  
2.10 Biological Endpoints  
Change in plasma IL-6, angiopoietin-2, cell-free DNA, syndecan-1, and cell-free hemoglobin 
between enrollment (baseline) and Days 2 and 3.  
3. Trial Description  
3.1 Background  
3.1.1 Background-Sepsis and sepsis-related organ dysfunction  
Sepsis is a common condition with over 750,000 cases per year in the U.S. and an associated 
mortality of at least 18-38% [2, 3].  The primary cause of death in sepsis is organ dysfunction, 
including acute respiratory distress syndrome (ARDS), acute kidney injury (AKI) and 
cardiovascular dysfunction.  ARDS is a common complication of sepsis; in a large multicenter 
international study of ARDS, sepsis (pulmonary or extrapulmonary) was the most common 
cause of ARDS [4].  Mortality from sepsis-induced ARDS is higher than mortality from ARDS 
due to other causes [5]. AKI is also common in sepsis, occurring in greater than 50% of patients 
[6-8]; the range of morbidity associated with AKI is broad, including prolonged ICU length of 
stay, need for renal replacement therapy (RRT), and death. Furthermore, ARDS and AKI 
commonly occur together, suggesting overlapping pathophysiologic mechanisms.  The 
development of AKI in patients with ARDS increases mortality [9]. However, despite the 
magnitude of this clinical problem, there are still no specific therapies for sepsis other than 


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antimicrobials, and no specific therapies that reduce the severity of sepsis-associated ARDS, 
AKI or cardiovascular dysfunction [10].  
 
 3.1.2       Cell-free hemoglobin (CFH) and organ dysfunction in sepsis  
Sepsis causes alterations in the red blood cell membrane that lead to release of CFH into the 
circulation due to membrane damage [11-13].  In an animal model of cecal-ligation and puncture 
induced sepsis, CFH can be measured in the plasma and levels of CFH are associated with 
mortality and worse renal function, an effect that is potentiated by deficiency of haptoglobin, 
hemopexin, or heme oxygenase-1[14]. In critically ill adults with sepsis, CFH is present in 80%-
90% of patients, is associated with oxidative injury as measured by plasma F2-isoprostanes (F2-
IsoP), and is independently associated with an 
increased risk of in-hospital mortality [15, 16] 
(Figure 1).    
Mechanisms of toxicity of CFH in sepsis include 
the ability of CFH to scavenge nitric oxide, 
leading to vasoconstriction in various vascular 
beds [17-20], proinflammatory effects of CFH 
and its heme subunit [21-23] and the ability of 
CFH to undergo redox cycling, leading to 
oxidation of lipid membranes and release of F2-
IsoP [24, 25]; plasma levels of F2-IsoP, are 
increased in patients with sepsis and are 
associated with renal, hepatic, and coagulation 
failure, and mortality [26]. This trial will test one 
agent, Acetaminophen, that can specifically 
target the injurious effects of CFH, present in 
the vast majority of patients with sepsis, to 
reduce organ dysfunction and affect clinical 
outcomes. To determine whether pre-
enrollment point-of-care measurement of plasma CFH can be used for predictive enrichment in 
a future phase 3 trial of this agent, samples will be collected at enrollment in the current trial for 
post hoc measurement of plasma CFH to determine what level of pre-enrollment plasma CFH is 
optimal for predictive enrichment.    
Figure 1.  Plasma  cell - free hemoglobin levels are  
significantly associated with in-hospital mortality in 391  
critically ill patients with sepsis (14)  


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3.2     Acetaminophen—Preclinical and Clinical Data  
The systemic inflammatory response to infection in sepsis creates an oxidative environment that 
has the potential of oxidizing the iron in CFH from the Ferrous (Fe 2+) to the ferryl (Fe 4+) state, 
creating an iron radical capable of organ injury 
[15, 25, 27]. Acetaminophen, at normal 
therapeutic concentrations, is a potent and 
specific inhibitor of hemoprotein-mediated lipid 
peroxidation owing to its ability to reduce the 
ferryl-protoporphyrin radical generated with 
release of hemoproteins into the circulation 
(Figure 2) [27, 28].  The hemoprotein reductant 
activity of Acetaminophen is independent of its 
other pharmacologic mechanisms, including 
prostaglandin H2 synthetase and 
cyclooxygenase inhibition [28, 29]. In an 
experimental study in rats, Acetaminophen was  
highly effective in preventing oxidative damage 
and renal failure in the setting of myoglobinemia due to rhabdomyolysis  [27].  In an 
observational study of patients with sepsis, receipt of Acetaminophen was associated with 
decreased F2-IsoP levels and independently associated with decreased in-hospital mortality 
[15].  
In the isolated perfused human lung, addition of CFH to the perfusate causes acute lung injury 
manifested by an increase in lung vascular permeability and pulmonary edema formation as 
measured by lung weight gain and increased alveolar capillary barrier permeability to protein as 
measured by accumulation of Evans Blue labeled albumin in the airspaces of the lung.  In this 
human lung preparation, addition of Acetaminophen to the perfusate blocked the effects of CFH 
on vascular permeability and pulmonary edema formation (Figure 3).  
 
Figure  6 .  Addition of  Acetaminophen  ( APAP) to the perfusate blocks the effects of CFH on pulmonary edema formation  
( panel A) and alveolar capillary barrier permeability (panel B) in paired isolated perfused human donor lungs.  
Figure 5 . Release of cell free hemoglobin into an oxidizing  
environment as in sepsis, leads to formation of the highly  
reactive Fe4+ ferryl radical 


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The Ware group conducted a phase 2a, randomized, double blind, placebo-controlled pilot 
clinical trial of enteral Acetaminophen in 40 critically ill adults with severe sepsis [30] entitled the 
ACROSS I Trial (Acetaminophen for the Reduction of Oxidative injury in Severe Sepsis).  
Patients were enrolled within 24 hours of admission to the ICU.  After enrollment, blood was 
drawn for measurement of plasma CFH to identify patients who may benefit from  
Acetaminophen therapy.  Patients with undetectable CFH (<10 mg/dL) were not randomized,  
while patients with any amount of CFH (≥ 10 mg/dL) were randomized to enteral  
Acetaminophen at the dose of 1 gram enterally every 6 hours or placebo for 3 days.  Notably, 
91% of patients who met inclusion criteria for this trial had detectable CFH and thus were 
eligible for randomization.  The primary endpoint was plasma F2-IsoP on study day 3, with 
secondary endpoints of F2-IsoP on study day 2 and serum creatinine on study day 3.  
On study day 3, F2-Isoprostanes were not significantly lower in the Acetaminophen group (30 
pg/mL, IQR 24-41) compared with placebo (36 pg/mL, IQR 25-80, p = 0.35) although power was 
reduced at that timepoint due to patient dropout.  However, F2-Isoprostanes were significantly 
reduced on study day 2 in the Acetaminophen group (24 pg/mL, IQR 19 – 36) compared with 
placebo (36 pg/mL, IQR 23-55, p = 0.047).  Creatinine on study day 3 was significantly lower in 
the Acetaminophen group (1.0 mg/dL, IQR 0.6–1.4) compared with placebo (1.3 mg/dL, IQR 
0.83 – 2.0, p = 0.039).  This difference persisted beyond study completion (Figure 4A). There 
was no statistically significant difference in hospital mortality (Acetaminophen 5.6% vs. placebo 
18.2%, p = 0.355) although there was a trend for benefit (Figure 4B).  There were no significant 
adverse events (AST or ALT >400, Acetaminophen 9.5% vs. placebo 4.3%, p = 0.599).   
  
Although the reduction in biomarkers of oxidative injury in the study was not consistent across 
all analyses, the biologic plausibility of benefit with Acetaminophen in patients with sepsis and 
detectable CFH, ease and safety of administration, consistent although non-significant 
improvement across all clinical outcomes, and the finding of improved renal function in sepsis 
warrants a larger Phase 2b trial in sepsis patients.  A recent clinical trial of oral Acetaminophen 
in severe falciparum malaria also showed a reduction in creatinine in the treated arm with no 
adverse safety signal, further supporting the need for larger clinical trials [31].    A phase 3 trial 
of acetaminophen for treatment of severe falciparum malaria in the Democratic Republic of 
Congo (NCT04251351) began enrolling in December 2021.  
 
 
Panel A: 
 
Figure 7, Panel A: Creatinine measured at enrollment, each study day, and after study completion in patients never requiring 
renal replacement therapy. Panel B: Hospital survival by treatment group. 
Panel B: 


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Another trial, the ANZICS HEAT trial [32], has some superficial similarities to the 
Acetaminophen arm of the ASTER trial but did not target sepsis patients with organ dysfunction 
and used relatively low cumulative doses of Acetaminophen. In the HEAT trial, critically ill 
patients (n = 690) with fever were randomized to receive IV Acetaminophen or placebo 1g every 
6h for up to 28 days, but only while febrile, leading to a median total dose of only 8g. This dose 
is significantly lower than our proposed total dose of 20g, which was chosen to provide 
continuous dosing over the early phase of sepsis during the time period when high levels of 
plasma CFH are present. Importantly, there was no signal for harm from Acetaminophen in the 
HEAT trial in terms of liver or other organ dysfunction or mortality. In summary, the HEAT trial 
does not provide any information about the efficacy of extended dosing of intravenous 
Acetaminophen in the target population that is of interest in ASTER and is relevant primarily 
with regards to safety of Acetaminophen in critical illness.  
We will use the FDA-approved dose and frequency 
of intravenous Acetaminophen (1g every 6h or 
4g/day, total dose 20 g; if weight < 50kg then dose 
reduced to 15 mg/kg every 6h). There are extensive 
safety data for Acetaminophen in healthy subjects 
receiving this dose and frequency enterally [33, 34]. 
Specifically, at this dose/frequency: (1) markers of 
liver injury are rarely elevated; (2) liver injury has 
not been observed until after 7 days of 
Acetaminophen use; and (3) any elevation of liver 
injury markers resolved after stopping the drug [33, 
35]. Compared to enteral Acetaminophen, IV 
administration reduces initial hepatic exposure by 
approximately 2-fold through lack of first-pass 
metabolism [36, 37]. This dose of 1g IV every 6 
hours was safe in 346 critically ill patients in the 
Acetaminophen arm in the HEAT trial with some patients receiving Acetaminophen at this dose 
for as long as 3 weeks [32]. In ASTER, the proposed duration of treatment is 5 days, rather 
than the 3 days studied in ACROSS. Treatment duration has been extended because our 
preliminary studies in critically ill patients enrolled in an ICU feeding trial [38], showed that high 
levels of plasma CFH persisted at 6 days (Figure 5). Acetaminophen should continue to be 
protective as long as the oxidative stimulus (CFH) is present; however, we elected to limit 
Acetaminophen treatment to 5 days to minimize any risk of hepatic injury.   
3.4 Summary  
Sepsis is common, associated with organ failure, high mortality, and currently without effective 
therapeutic interventions directed at the underlying pathophysiology. Acute lung injury (ARDS), 
acute kidney injury and cardiovascular dysfunction are common organ injuries in sepsis, are 
associated with increased mortality, and may be mediated, in part, by the elevated levels of 
CFH and pro-oxidant pathways that are characteristic of sepsis. Acetaminophen, a potent 
inhibitor of CFH-mediated oxidative injury, improves lung and renal function in pre-clinical 
models and seems to be potentially beneficial in humans with hemoprotein-mediated diseases, 
including in critically ill adults with sepsis.  
Figure 8 .  Comparison of plasma CFH  
levels in the ACROSS study (days 0,1,2,3)  
and the Vanderbilt phase 2 EDEN study  
( days 0, 6), a trial of different caloric intake  
goals in critically ill patients  


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ASTER was originally designed as  an efficient platform for conducting two phase 2b trials with 
a shared placebo group in which we will assess the potential impact of Acetaminophen and 
Vitamin C on reducing organ injury in a population of patients with sepsis who have been 
prognostically enriched with evidence of either cardiovascular dysfunction or respiratory 
insufficiency (or both). The Vitamin C arm was stopped on June 15th and the trial will resume as 
a trial of Acetaminophen vs placebo.  The trial will provide information on both the primary and 
secondary endpoints, including the biological endpoints, that will guide decisions in the future to 
proceed to larger phase 3 trials that would be focused on 90-day mortality. In addition, the 
results of this trial will establish whether a point-of-care measurement of the presence of cell-
free hemoglobin should be used as a predictive enrichment strategy for selecting patients for 
future clinical trials of Acetaminophen. Also, it is important to note that intravenous 
Acetaminophen is readily available and inexpensive, increasing the potential impact of this trial.    
4. Trial Design  
4.1 Study Description  
Prospective multi-center phase 2b randomized placebo-controlled double-blinded interventional 
trial of intravenous Acetaminophen for patients with sepsis-induced hypotension or respiratory 
failure. Patients greater than or equal to 18 years old with known or suspected infection with 
associated hemodynamic or acute respiratory dysfunction will be eligible for enrollment.    
Only subjects who are admitted or planned to be admitted to an intensive care unit (ICU) will be 
enrolled. Study drug administration will occur for a total of 120 hours, unless one of the following 
occurs:   
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
5. New AST or ALT elevation greater than 10 times the upper limit of normal level  
A total of 450 participants who meet all of the inclusion criteria and none of the exclusion criteria 
will be randomized in a 1:1 fashion ( Acetaminophen:placebo) into the two treatment arms of the 
study.  The two treatment arms are: Acetaminophen (225 patients) and Placebo (225 patients). 
Enrollment will occur over approximately 15 months.  
 


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The study interventions are:  
  
 
 
 
The safety and efficacy will be evaluated through repeated laboratory assessment, clinical 
assessments and biomarker measurements during the entire study. Adverse events and clinical 
outcomes, including need for assisted ventilation, vasopressors, renal replacement therapy, ICU 
length of stay, and hospital length of stay, will be assessed through 28 days or hospital 
discharge, whichever occurs first. Hospital mortality and all-cause mortality at 90 days will also 
be assessed.    
4.2 Randomization  
After informed consent and completion of screening, we will randomize each eligible participant 
using a web-based randomization system. Designated research team members will have 
permission to randomize participants. At randomization, each subject will be assigned a unique 
randomization ID number that the site pharmacy can use to determine study arm assignment to 
Acetaminophen or placebo  
The CCC will send a confirmation email to the randomizer containing the randomization ID 
number. We will stratify randomization by enrolling hospital.  
4.3 Primary Efficacy Variable 
Primary efficacy variables (days alive and free of assisted ventilation, vasopressors, and new 
renal replacement therapy) will be evaluated comparing Acetaminophen vs. placebo: over the 
first 28 days Patients need to be free of all three components to qualify for a day alive and free 
from organ failures.  A patient could be free of all organ failures from day 10 through day 22 and 
then die on day 22 and would receive 12 days of credit for this primary outcome. The 
components of this outcome (days alive and free of assisted ventilation, vasopressors and new 
renal replacement therapy in the overall cohort and in survivors, and 28-day hospital mortality) 
will also be reported.    
1. Acetaminophen given intravenously at the dose of 1 gram (or 15 mg/kg if patient 
weighs < 50 kg) every six hours for 5 days (20 doses) OR 
2. Placebo (identical appearing 5% dextrose solution) infused every six hours for 5  
days (20 doses)  


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 
This three-component primary outcome was chosen as preliminary data suggest the 
study intervention has the potential to reduce mortality and favorably impact at least two 
of the three organ failure support outcomes, thus increasing the sensitivity of this 
outcome for benefit in a Phase 2b trial designed primarily to assess safety and proof of 
concept [39, 40].  
4.4 Secondary Clinical Efficacy Variables 
The ventilator, vasopressor, new renal replacement, and hospital “free days” endpoints below 
are traditional Sepsis and ARDS Phase 2 and Phase 3 composite outcomes that assign a value 
of zero free days for participants dying on or before study day 28 to control for the competing 
risk of death as described in Section 8.3 of the protocol.  These traditional outcomes are 
included in this Phase 2b trial to search for additional evidence of efficacy that could support a 
decision to progress to Phase 3 and will allow for comparison to other sepsis and ARDS trials 
using these endpoints, including all other PETAL studies.   
1. 28-day ventilator-free days  
2. 28-day vasopressor-free days  
3. 28-day new renal replacement-free days  
4. 28-day hospital mortality   
5. 28-day ICU free-days  
6. 28-day hospital-free days to discharge home  
7. 28-day duration of ICU stay in survivors and non-survivors  
8. Initiation of assisted ventilation to day 28  
9. Initiation of renal replacement therapy to day 28  
10. Change in organ-specific Sepsis-related Organ Failure Assessment (SOFA) scores 
between enrollment and study day 7 (Appendix B)  
11. 90-day hospital mortality (includes death at any healthcare facility that is not the 
equivalent of home for a patient)  
12. Development of ARDS within 7 days of randomization  
13. Change in serum creatinine from enrollment to discharge, death, initiation of dialysis 
or 28 days, whichever occurs first   
14. Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline, need for new renal replacement therapy, or death  
15. Change in the Radiographic Assessment of Lung Edema (RALE) score from 
enrollment to study day 3 in patients who are receiving assisted ventilation or high 
flow nasal oxygen at baseline.  
16. 90-day all-cause mortality   
NOTE: Please see section 8.3 for description of how these variables are defined and calculated.  


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4.5 Safety Variables 
Safety monitoring and analysis will be performed separately on the Acetaminophen and 
placebo arms. The safety variables will include the following:  
1. Allergic or allergic-like reactions such as urticarial drug rash, hyperemia with 
hypotension, and anaphylaxis   
2. AST and ALT measured on study days 0, 2-5, and 7 (day 7 measurement can be ± 1 
day)   
3. The trough plasma levels measured just prior to a study dose of Acetaminophen on 
Day 2 (subject must have received at least 5 doses of study drug before the day 2 
samples are obtained) in the first 100 patients will be reported to the DSMB to 
determine if they exceed the level of 20 micrograms/mL, the potential level for 
hepatotoxicity.  
4. Administration of fluid bolus, new use of vasopressor, or increased dose of 
vasopressor within 120 minutes of study drug infusion. 
5. Incidence of reported adverse events  
4.6 Biological Endpoints 
Change in plasma IL-6, angiopoietin-2, cell-free DNA, Syndecan-1, and cell-free hemoglobin 
between enrollment (baseline) and Days 2 and 3.  
5. Treatments Arms and Study Drug Administration  
All study drug will be administered by the bedside nurse to the patient through an indwelling 
peripheral or central venous access line.  Study drug will be held if the patient loses all 
intravenous access and will be restarted as soon as intravenous access is restored.  Missed 
doses will be recorded but will not be added on to the end of the study.  If the patient is 
discharged from the ICU prior to day 5 or 20 doses, study drug will be stopped. A list of drug 
incompatibilities for Acetaminophen will be provided to each clinical site.   
5.1 Acetaminophen Arm  
 
Treatment: IV Acetaminophen  
Dose: 1 gm (or 15 mg/kg if patient is less than 50 kg)  
Frequency: Every 6 hours  
Duration: 5 days (20 doses) or ICU discharge, whichever comes first  


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 Patients randomized to Acetaminophen will receive Acetaminophen (hospital pharmacy stock) 
intravenously at the dose of 1 gram (or 15 mg/kg if patient < 50kg) every 6 hours for a maximum 
of 5 days (20 doses). Intravenous Acetaminophen was chosen over enteral Acetaminophen to 
insure adequate drug delivery in critically ill patients. In the ACROSS trial of enteral 
Acetaminophen (1 g every 6 hours), the observed serum Acetaminophen levels were at the 
lower limit of the therapeutic window (Figure 9). Intravenous Acetaminophen is expected to 
provide higher peak levels of Acetaminophen 
while avoiding first pass metabolism by the liver. 
Based on the observation that cell-free 
hemoglobin levels remain elevated at 6 days in 
critically ill patients (see Figure 5, section 3.3), 
treatment will be continued for 5 days (for a 
maximum of 20 doses) or until any of the following 
occur:  
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
5. New AST or ALT elevation greater than 10 
times the upper limit of normal level.  
For blinding purposes, Acetaminophen will be 
transferred to a sterile INTRAVIA container 
(Baxter) or its equivalent prior to dispensing by the 
pharmacy. 
   
5.2 Placebo Arm  
 
Patients randomized to Placebo arm will receive an identical appearing room temperature 5% 
dextrose in water placebo intravenously every 6 hours for a maximum of 5 days (for a maximum 
of 20 doses). A similar placebo has been used in other randomized clinical trials of IV 
Acetaminophen. The =-Placebo will be 100 ml of 5% dextrose in water. Patients weighing <50 kg 
will receive a similar weight-adjusted volume to patients in the active treatment arm. Treatment 
will stop at 5 days (20 doses) or until any of the following occur:  
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
Figure 9.   Serum  Acetaminophen   levels on  
enrollment and each study day of the ACROSS  
trial of enteral Acetaminophen versus placebo in  
sepsis   
Treatment: Placebo 
Dose: 5% dextrose in water  
Frequency: Every 6 hours  
Duration: 5 days (20 doses) or ICU discharge, whichever comes first  


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4. Death;   
5. New AST or ALT elevation greater than 10 times the upper limit of normal level.   
5.3 Drug Interruptions  
Interruptions of study drug should be avoided. If the study drug must be interrupted due to loss 
of intravenous access, it should be restarted as soon as the treatment team re-establishes 
intravenous access.  In cases in which the study drug was interrupted, the missed doses will not 
be added on to the end of the study. If a dose cannot be administered within 3 hours of the 
scheduled time, the dose should be skipped.  Any dose administered outside of the +/- 1-hour 
window or any dose that is skipped will trigger a protocol deviation.  
Any study drug interruptions will be recorded, including the reason for the interruption.    
In the case of a new measurement of an AST or ALT greater than or equal to 10 times the 
upper limit of normal or rash or other hypersensitivity attributed to the study drug during the 
study period, the study drug will be stopped and not restarted. Elevation of AST or ALT is known 
to reverse with discontinuation of Acetaminophen [32]; in almost all cases simply discontinuing 
the study drug is appropriate. If study drug is discontinued for AST and/or ALT elevation we will 
repeat AST and ALT at 24 and 48 hours after the study drug is discontinued and continue to 
measure daily until the levels have declined from their peak level.    
6. Study Population and Enrollment  
 6.1 Participant Enrollment  
The study is expected to accrue over approximately 15 months. Participants will be recruited 
from the Emergency Department or Intensive Care Unit at PETAL Network centers. Participants 
who withdraw or prematurely discontinue study drug will be analyzed by an intention to treat 
analysis. Safety data will continue to be collected up to hospital discharge for any participants 
who are withdrawn from the study.  
6.2 Inclusion Criteria  
1. Age ≥ 18 years  
2. Sepsis defined as:  
a. Clinical evidence of a known or suspected infection and orders written to administer 
antibiotics  
AND  
b. Hypotension as defined by the need for any vasopressor (and 1 liter of fluid already 
administered intravenously for resuscitation) OR respiratory failure defined by 
mechanical ventilation, BIPAP or CPAP at any level, or greater than or equal to 6 
liters/minute of supplemental oxygen (criterion b must be present at the time of 
randomization)  
3. Admitted (or intent to admit) to a study site ICU within 36 hours of presentation to the 
study site ED or within 36 hours of presentation to any acute care hospital  


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6.3 Exclusion Criteria  
1. No consent/inability to obtain consent from the participant or a legally authorized 
representative  
2. Patient unable to be randomized within 36 hours of presentation to the ED or within 36 hours 
of presentation to any acute care hospital  
3. Diagnosis of cirrhosis by medical chart review  
4. Liver transplant recipient  
5. AST or ALT greater than five times the upper limit of normal   
6. Diagnosis of ongoing chronic alcohol use disorder/abuse by chart review; if medical record 
unclear, use Appendix F  
7. Hypersensitivity to Acetaminophen  
8. Patient, surrogate or physician not committed to full support (Exception: a patient will not be 
excluded if he/she would receive all supportive care except for attempts at resuscitation from 
cardiac arrest)  
9. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used only 
for sleep-disordered breathing   
10. Chronic dialysis 
11. Use of home oxygen > 3 liters/min nasal cannula for chronic cardiopulmonary disease  
12. Moribund patient not expected to survive 24 hours   
13. Underlying malignancy or other condition with estimated life expectancy of less than 1 month  
14. Pregnant woman, woman of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or woman who is breast feeding  
15. Prisoner  
16. Treating team unwilling to enroll because of intended use of Acetaminophen  
6.4     Reasons for each of the exclusion criteria  
1. No consent/inability to obtain consent – no justification for waiver of informed consent.   
2. Patient unable to be randomized within 36 hours of presentation: limits enrollment to acute 
conditions and presentations.  
3. Diagnosis of cirrhosis – not safe to administer 4 grams of IV Acetaminophen daily in this 
clinical setting  
4. Liver transplant recipients are excluded as the doses of acetaminophen used in this study 
exceed recommended doses for liver transplant patients   
5. AST or ALT greater than five times the upper limit of normal – this is a standard precaution 
to exclude patients with acute or chronic liver disease [38].   
6. Diagnosis of ongoing chronic alcohol use disorder/abuse – these patients may have an 
increased risk of hepatic toxicity from Acetaminophen.   
7. Hypersensitivity to Acetaminophen– self-evident.  


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8. ICU staff, legally authorized next of kin or study subject not committed for full support – 
required to assess the potential beneficial effects of Acetaminophen on the primary and 
secondary outcomes.  
9. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used 
only for sleep-disordered breathing – these patients would have ventilator dependent 
respiratory failure.  
10. Chronic dialysis is excluded because one component of the primary end-point is days free of 
dialysis over the first 28 days in the trial. 
11. Use of home oxygen > 3 liters/min nasal cannula for chronic cardiopulmonary disease- these 
patients with severe  chronic respiratory failure may not  be a good candidate for an 
intervention designed in part to facilitate lung injury resolution.  
12. Moribund patient not expected to survive 24 hours – these patients would not fit the goals of 
testing an intervention designed to reduce organ failure from sepsis.  
13. Underlying malignancy or other condition with estimated life expectancy of less than 1 month 
- these patients would not match with the secondary outcome of testing the beneficial effects 
of this therapy on 28-day mortality.   
14. Pregnant female, female of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or female who is breast feeding – 
safety not established for using the intervention to be tested in this trial.  
15. Prisoner – not correct to include these patients in clinical trials by ethical standards.  
16. Treating team unwilling to enroll because of intended use of Acetaminophen– open label 
Acetaminophen use is prohibited in this trial because the study drug dose is the maximum 
FDA approved dose and would also confound interpretation of the results of the trial.  
6.5 Co-enrollment 
Co-enrollment in other clinical trials will only be allowed after a trial has been approved for co-
enrollment by ASTER study leadership and the PETAL DSMB. Interventions that are 
contraindicated in combination with acetaminophen are not permitted.  Study procedures of the 
co-enrolling trial must not impose an undue burden on research participants or research staff 
when viewed within the context of ASTER study procedures, including the total volume of blood 
drawn for research and follow-up activities. For blinded trials, the co-enrolling trial must agree to 
confidentially provide unblinded treatment assignments to the respective DSMBs and the 
unblinded Medical Monitors to allow for accurate assessment of serious adverse events and 
unanticipated problems. 
6.6 Informed Consent  
Screening will occur in the Emergency Department and intensive care units daily to identify 
potential candidates for enrollment. Once a potential participant is identified, the patient’s 
primary medical team will be contacted to secure permission to approach the patient or their 
legally authorized representative regarding possible participation in the study.  If the primary 
team agrees, the study coordinator or investigator will meet with the patient and/or legally 
authorized representative and will describe the proposed study protocol in lay terminology.  
Prior to performing any study procedures, an informed consent form (ICF) approved by the 
central IRB will be signed and dated by each study participant or legally authorized 
representative and the study investigator or person informing the subject and obtaining the 


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consent. Prior to taking part in the study, the participant or his or her legal representative will 
receive a copy of the signed and dated ICF.    
A special vulnerable population, cognitively impaired patients, will be eligible for enrollment in 
this proposed study. Many septic patients have cognitive impairment as part of their acute 
disease process and others require sedation during their treatment course. In cases where the 
potential participant is cognitively impaired, surrogate consent will be obtained from the 
participant’s legally authorized representative according to local state law. In these cases if the 
cognitive impairment has resolved prior to hospital discharge, the participant will be reconsented 
at the earliest opportunity and given the option of continuing or stopping their participation.    
6.7 Excluded Medications  
Clinicians will be prohibited from administration of Acetaminophen (either intravenously or 
enterally) to study participants during the study.  Additionally, administration of open label 
Acetaminophen at any dose will be recorded in the case report forms.   
6.8 On-study Fever Management Recommendations  
Use of nonsteroidal anti-inflammatory drugs (NSAIDs) or Cyclooxygenase-2 inhibitors to treat 
fever and pain will be discouraged during the 5-day study drug administration and the use of 
these medications for other indications will also be discouraged. No restriction will be placed on 
the use of aspirin in doses of less than or equal to 325 mg per day.   
The use of open-label acetaminophen will not be permitted while receiving study medication. 
Once the course of study drug administration has been completed, open-label acetaminophen 
can be administered at the discretion of the treating clinician. Physical cooling will be used at 
the discretion of the clinical team per local practice preferences. 
This plan will apply to both the Acetaminophen and placebo arms Fever Management 
Recommendations are also outlined in the Appendix.  
6.9 Concomitant Medications  
We will record use of non-steroidal anti-inflammatory agents, iodinated contrast, glucocorticoids 
(all preparations including hydrocortisone, methylprednisolone, prednisone, dexamethasone), 
thiamine, remdesivir, baricitinib, tocilizumab. 
6.10 Common Strategies for Study Participants  
6.10.1 Ventilator Procedures  
Low tidal volume ventilation and weaning from mechanical ventilation will follow standard clinical 
protocol for participants meeting criteria for ARDS (Appendix C).  
Low Tidal Volume Ventilation: We will use a simplified version of the ARDS network 6 ml/kg 
PBW lung protective ventilation protocol. If not already being used, a low tidal volume protocol 
for mechanical ventilation must be initiated within two hours of randomization in all patients who 
meet criteria for ARDS.   


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Weaning: Since the time at which a patient achieves unassisted ventilation affects some 
secondary endpoints, and because recent evidence-based consensus recommendations have 
identified a best practice for weaning, a weaning strategy will also be controlled by protocol 
rules in accordance with these evidence-based recommendations. This weaning strategy is a 
simplified version of the protocolized weaning strategy used in prior PETAL and ARDS Network 
studies.  
6.10.2   On Study Fluid Management  
Fluid management during shock will be unrestricted. Balanced crystalloid solution (such as 
lactated Ringers, Hartman’s solution, Plasmalyte or Normosol) is preferred, for resuscitation. 
Given the lack of clear evidence identifying a superior approach, the type of fluid administered 
will not be mandated.   
However, in patients not in shock or after shock resolution, a conservative fluid approach will be 
recommended for patients with ARDS. This conservative fluid management approach is a 
simplification of the algorithm utilized in the ARDS Network FACTT study (Appendix D).   
6.12 Criteria for Discontinuation or Withdrawal  
The primary reason for treatment discontinuation will be recorded using the following categories:  
a. Adverse Event: The participant has experienced an adverse event that the investigator 
believes requires early termination because continued participation imposes an 
unnecessary risk to the participant’s health.  
b. Major Protocol Deviation: The participant failed to meet protocol entry criteria or did not 
adhere to protocol requirements, excluding prolonged interruption of study drug.  
c. Voluntary Withdrawal: The participant or legally authorized representative wishes to 
withdraw from the study. The reason for withdrawal, if provided, will also be documented in 
the CRF.  
d. Other: This category includes participants withdrawn by the treating team for a new 
indication for Acetaminophen use.  
7.0 Study Plan  
7.1 Baseline Assessments  
The following will be collected from information available prior to randomization. If more than 
one value is available in the 24 hours prior to randomization, the value closest to the time of 
randomization will be recorded.    
1. Demographic and Admission Data (including age, sex, race)  
2. Pertinent Medical History and Physical Examination (including components of the 
Charlson co-morbidity score)  
3. Height; gender; measured Body Weight (mBW); calculated predicted body weight 
(PBW); body mass index (BMI)  


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4. Location when inclusion criteria met: ED, ICU  
5. Vital signs:  heart rate (beats / min), systemic systolic and diastolic BP (mmHg), body 
temperature (°C), and oxygen saturation with FiO2  
6. Use of vasopressors, corticosteroids, acetaminophen, thiamine, supplemental O2, 
oxygen saturation, invasive or non-invasive/invasive mechanical ventilation and renal 
replacement therapy prior to randomization   
7. Clinical laboratory testing results (AST, ALT, total bilirubin, serum bicarbonate)  
8. Serum creatinine: 
a 
Prehospitalization: Most recent outpatient creatinine from 24 hours to 365 days 
prior to admission if available 
b 
Baseline: Lowest in hospital creatinine prior to randomization  
9. Chest radiograph   
10. ARDS assessment if receiving assisted ventilation  
11. Presumed site of primary infection  
12. SOFA Score: as described in Appendix B  
13. Glasgow Coma Score 
14. Cell free plasma hemoglobin (to be collected and measured in batch at the end of the 
study)  
15. COVID-19 status and date of first positive test if known within 3 weeks of enrollment 
 7.2. Assessments During Protocol Phase (through study day 7)  
1. Daily fluid administration for the first 7 days (truncated for death or ICU discharge)  
2. Daily fluid outputs for the first 7 days (truncated for death or ICU discharge)  
3. Chest radiograph on day 3 (± 1 day) for all patients who were receiving assisted 
ventilation or high flow nasal oxygen at the time of randomization if they are still in the 
study hospital  
4. ARDS assessment through study day 7 (± 1 day). We will determine the presence and 
severity of ARDS for each day of assisted ventilation to day 7 using the following 
approach.  For participants on assisted ventilation with P/F <300 or S/F <315, FiO2 
≥40%, and PEEP ≥5 cm H2O, determine if hypoxemia is valid, acute, and not fully 
explained by CHF or fluid overload. If yes, local investigators will review the first CXR 
(or CT) performed on each ventilated day with valid P/F or S/F <315 (to day 7). If no 
chest imaging studies are present that day, site investigators may review available 
imaging one day before or after to determine if ARDS imaging criteria met. ARDS 
imaging criteria are met if the images are consistent with ARDS (bilateral opacities not 
fully explained by effusions, lobar/lung collapse, or nodules). If equivocal, the reviewing 
investigator will adjudicate with additional investigators.   


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6. AST and ALT, and bilirubin on days 1-7 if available.   AST/ALT REQUIRED on days 2-
5, and 7 (day 7 measurement can be ±1 day). Daily SOFA score through day 7 or 
hospital discharge   
7. Recording of fluid bolus (500 mL or more), new vasopressor administration, or increase 
of vasopressor dose within 120 minutes of receiving study drug  
8. Administration of the following concomitant medications: non-steroidal anti-inflammatory 
agents, iodinated contrast, glucocorticoids, thiamine, remdesivir, baricitinib, and 
tocilizumab   
7.3. Other Assessments and Data Collection  
1. Vasopressor use over 28 days  
2. Study drug administration 
3. Use of invasive and non-invasive ventilation (except noninvasive ventilation solely for 
sleep disordered breathing) over 28 days  
4. New renal replacement therapy over 28 days  
5. Last creatinine prior to initiation of RRT, death, study hospital discharge, or day 28 
(whichever comes first)  
6. Highest creatinine between randomization and day 28.  
7. Results of microbiologic testing from samples taken from up to 24 hours prior to 
randomization until 72 hours after randomization 
8.  Source of infection  
7.4. Assessments after Hospitalization  
We will perform an assessment using all testing and records available during the hospitalization 
where a site investigator will make an assessment as to whether the hypotension at enrollment 
was likely due to an infection or not.  
7.5. 90-day All-Cause Mortality  
We will contact patients at day 90 to ascertain their survival status.  This will be done by 
telephone contact with the patient or family members as well as a review of medical records.  
We will use publicly available data sources and the national death index as a final check for 
patients with whom we are unable to confirm their vital status through other means.  
7.6. Specimen Collection  
Plasma: Blood will be collected within 2 hours of randomization (prior to first study drug 
administration) and on days 2 and 3 (approximately 36 mls total).  To minimize hemolysis, blood 
will preferentially be collected through central venous access devices if present. If there is no 
central venous access, then either an arterial or peripheral venous stick can be used, with blood 
drawn slowly to minimize hemolysis. The blood samples will be centrifuged, and the plasma will 
be frozen and stored at a biorepository for future research for the following purposes:  


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• 
Biological Endpoint Assays: To fulfill the secondary biological endpoints, the plasma 
samples will be measured for the concentration of plasma IL-6, angiopoietin-2, 
Syndecan-1, and cell-free DNA on enrollment and on days 2 and 3 and analyzed for 
significant changes from baseline in the placebo and  Acetaminophen groups.  
• 
Additional Assays: In addition, the plasma samples at baseline and on days 2 and 3 will 
be used to measure cell-free hemoglobin.  
• 
Acetaminophen Levels: Plasma collected as a trough level just prior to a study dose on 
Day 2 (subject must have received at least 5 doses of study drug before the day 2 
samples are obtained) will be used to measure Acetaminophen levels in the first 100 
patients randomized to the Acetaminophen arm.  Measuring Acetaminophen levels in 
critically ill patients with sepsis will address a knowledge gap in the literature and 
contribute to interpretation of the results of the trial and also provide data that the DSMB 
can review after 100 patients are enrolled in the Acetaminophen arm. For the first 100 
patients in the acetaminophen arm, the blood drawn on day 2 (between 8 am and 2 pm) 
will be timed to occur 30 minutes before the next study drug administration (for a trough 
level of acetaminophen).   
• 
Samples for Future Research: Plasma that has not been used for these planned 
assays will be stored in a biorepository for future research funded by ancillary studies.  
Whole Blood for Genetics: When consent for genetic testing is specifically obtained, additional 
whole blood for future RNA and DNA studies will be collected at baseline and on day 3. Total 
blood volume for the genetic samples is approximately 8.5 ml/day, or 17mls total. Total overall 
blood volume collected (plasma and DNA/RNA) is approximately 53 ml.   
Urine: Urine will be collected within 2 hours of randomization and on days 2 and 3 (if patient still 
in the ICU), frozen and stored along with plasma. Urine samples will be saved for ancillary 
studies.  
Storage and Shipment: All samples collected in this trial will be identified by a coded number 
during shipment and storage at the central biorepository. 
8. Statistical Methods  
8.1 Primary Endpoint  
The primary endpoint of this trial is to evaluate the effects of intravenous Acetaminophen 
compared to placebo on the number of days alive and free of assisted ventilation, vasopressors 
and new renal replacement therapy between the treatment arm and the placebo group over the 
first 28 days.   
Note that the primary outcome components to be reported will be:  
1. 28-day mortality prior to discharge home   
2. Days free of assisted ventilation to day 28 in the overall cohort and survivors  
3. Days free of new RRT to day 28 in the overall cohort and survivors  
4. Days free of vasopressors to day 28 in in the overall cohort and survivors  


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The primary safety objective is to determine the effect of intravenous Acetaminophen on markers 
of liver injury (AST and ALT) measured on days 2-5, and 7 (day 7 measurement can be ±1 day).  
8.2   Sample Size  
The primary study outcome is 28-day organ support free days (alive and without assisted 
ventilation, new renal replacement and vasopressors). Using clinical trial data from the PETAL 
Network, the estimated standard deviation of 28-day organ support free days (obtained from the 
PETAL Network CLOVERS study) is 8.7 days. With a sample size of 225 in the  active 
treatment group, and 225 in the r placebo treatment group, a two sample Z-test will have 85% 
power at a two-sided alpha level of 0.05 to detect a significant difference between active 
treatment and placebo if the true mean difference is 2.5 support free days. .We will target a total 
enrolment of 450 participants to offset a 2% rate of loss to follow up. 
8.3 Secondary endpoints  
We will compare the efficacy of intravenous Acetaminophen to placebo for efficacy endpoints.  
We will compare the safety of Acetaminophen to placebo for safety endpoints.  
• 
28-day ventilator-free days: VFDs depend on both duration of ventilation and mortality 
through study day 28. In participants who survive 28 days, VFD is defined as 28 minus 
days of invasive or noninvasive ventilation to day 28.  Duration of ventilation is counted 
from the first study day of assisted breathing through the last day of assisted breathing 
provided the last day is prior to day 28. For participants discharged with assisted 
ventilation (e.g., to LTAC facility) prior to day 28, a phone call will be required to assess 
ventilator and vital status at day 28. Participants discharged prior to day 28 on 
unassisted breathing will be assumed to remain on unassisted breathing through day 28. 
Isolated periods of ventilation briefer than 24 hours for surgical procedures and 
ventilation solely for sleep disordered breathing do not count towards duration of 
ventilation. In participants who never require assisted breathing, duration of ventilation is 
zero. Participants who do not survive 28 days will be assigned zero VFD.  
• 
28-day vasopressor-free days: Vasopressor free days to day 28 are defined as the 
number of calendar days between randomization and 28 days later that the patient is 
alive and without the use of vasopressor therapy.  Patients who die prior to day 28 and 
those who receive vasopressor therapy for the entire first 28 days are assigned zero 
vasopressor free days.    
• 
28-day new renal replacement-free days: Renal replacement free days to day 28 are 
defined as the number of calendar days between randomization and 28 days later that 
the patient is alive and without renal replacement therapy. We also follow the “last off” 
method. Patients who died prior to day 28 and those who receive renal replacement 
therapy for the entire first 28 days are assigned zero renal replacement free days.  
• 
28-day hospital mortality  
• 
28-day ICU free days:  ICU free days to day 28 are defined as the number of days 
spent alive and out of the ICU to day 28.   
• 
28-day hospital free days to discharge home: Hospital free days to day 28 are 
defined as 28 days minus the number of days from randomization to discharge home. If 
a patient has not been discharged home prior to study day 28 or dies prior to day 28, 


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hospital free days will be zero. Patients transferred to another hospital or other health 
care facility will be followed to day 28 to assess this endpoint.  
• 
Duration of ICU stay in survivors and non-survivors: The total number of days spent 
in the ICU until hospital discharge or death during the first 28 days. If a patient is 
discharged alive from the study hospital we assume they are no longer in the ICU.  
• 
Initiation of assisted ventilation to day 28:  Any patient who received assisted 
ventilation in the first 28 days meets this endpoint. If a patient leaves the hospital without 
initiation of assisted ventilation, we assume they never started.  
• 
Initiation of new renal replacement therapy to day 28.  
o Patients who receive (new) renal replacement therapy through day 28 will meet 
this endpoint.  
• 
Change in organ-specific Sepsis-related Organ Failure Assessment (SOFA) scores 
between enrollment and day 7: We will calculate the SOFA score upon enrollment and 
at day 7 using clinically available data. If a value is not available at baseline, it will be 
assumed to be normal. At the day 7 assessment, if a value is missing then we will carry 
forward the closest previously known value. The GCS component of the SOFA score will 
be omitted.  (See Appendix B)  
• 
90-day hospital mortality (any hospital)  
• 
Development of ARDS within 7 days: We will determine the presence and severity of 
ARDS for each day of assisted ventilation to day 7. For participants on assisted  
ventilation with P/F <300 or imputed P/F <300, FiO2 ≥40%, and PEEP ≥5 cm H2O, 
determine if hypoxemia is valid, acute, and not fully explained by CHF or fluid overload. 
If yes, local investigators will review the first CXR (or CT) performed on each ventilated 
day with valid P/F or imputed P/F <300 (to day 7). If no chest imaging studies are 
present that day, site investigators may review available imaging one day before or after 
to determine if ARDS imaging criteria are met. ARDS imaging criteria are met if the 
images are consistent with ARDS (bilateral opacities not fully explained by effusions, 
lobar/lung collapse, or nodules). If equivocal, the reviewing investigator will adjudicate 
with additional investigators.  
• 
Change in serum creatinine from enrollment to discharge, death, initiation of dialysis, or 
day 28, whichever occurs first.  
• 
Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline (defined as most recent outpatient creatinine from 24 
hours to 365 days prior to admission if available: if not available, lowest in hospital 
creatinine prior to randomization), need for renal replacement therapy, or death  
• 
Change in Radiographic Assessment of Lung Edema (RALE) score from enrollment to 
study day 3 in patients who are receiving assisted ventilation or high flow nasal oxygen 
at the time of study randomization  
• 
The safety of intravenous Acetaminophen compared to placebo as measured by daily 
serum AST, ALT, blood pressure following study drug administration (both arms) and 
incidence of reported adverse events (both arms)  


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• 
Change in plasma IL-6, plasma IL-8, plasma angiopoietin-2, plasma cell-free 
hemoglobin, syndecan-1, and plasma cell-free DNA from enrollment to days 2 and 3  
• 
90 day all-cause mortality: We will contact patients at day 90 to ascertain their survival 
status. This will be done by telephone contact with the patient or family members as well 
as a review of medical records and publicly available data sources. We will use the 
national death index as a final check for patients with whom we are unable to confirm 
their vital status through other means.  
8.4 Safety Endpoints  
AST and ALT will be measured on days 2, 3, 4, 5, and 7 after study enrollment. We will also 
collect data daily on potential adverse reactions to Acetaminophen, such as rash and 
hypersensitivity.  If either are present and thought to be due to the study drug, the study drug 
will be stopped.  
8.5 Predictive Biomarker  Analysis  
We will evaluate the predictive value of baseline plasma CFH for identifying patients who have 
an improvement in the primary endpoint, the three components of the primary endpoint, 28-day 
mortality or improvement in renal function as assessed by MAKE28. 
8.6 Statistical Analysis Plan  
The principal analyses will be intention-to-treat (based upon randomization assignment). A 
comprehensive statistical analysis plan will be written prior to the un-blinding of the primary and 
secondary efficacy results.  
8.6.1 Sub-Group by Treatment Interaction 
We will do a sub-group by treatment interaction analysis comparing outcomes in acute COVID-
19, defined as a positive diagnostic test for SARS CoV-2 in the prior three weeks and 
pneumonia as primary site of infection, and COVID-19 negative patients for the primary and 
secondary efficacy endpoints. The analysis will include a tabulation of the treatment effect in 
COVID-19 positive patients, the treatment effect in COVID-19 negative patients, and the 
difference between the treatment effect in these two sub-groups. We will also apply this 
interaction analysis separately to two additional pre-randomization factors, received 
acetaminophen (yes versus no), and home oxygen requirement (yes versus no).  
8.7 Interim Analysis  
The DSMB will have the ability to analyze all safety data, including AST/ALT, at any time during 
the study and stop the study indefinitely for safety reasons. The safety report will include a 
tabulation of all baseline demographics and physiologic variables as well as on-study variables, 
including SOFA scores, and all adverse events by randomized groups. For monitoring safety, 
data on 28-day mortality will be included with 95% confidence intervals for any differences 
between placebo and the acetaminophen group. In addition, a formal safety analysis will be 
done after 225 patients are accrued. This report will include an analysis of acetaminophen 
levels.  Trough levels greater than 20 mcg/mL will be evaluated for any relationship to AST/ALT 
levels.  Investigators will recommend protocol modification as needed to the DSMB as part of 
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Based on an interim analysis of the primary efficacy variable, this trial may stop for efficacy or 
futility of APAP relative to placebo. The stopping rules are based on two stage Haybittle-Peto 
stopping boundaries [42, 3] with Z=3.4 for early efficacy stopping. We plan one interim look at 
approximately one half of total enrollment and a final look at full enrollment.   
Table 1 below presents the stopping boundaries as a function of the mean difference in the 
primary efficacy variable favoring active treatment and the one-sided p-value favoring active 
treatment. Tables 2 and 3 present the probability of stopping at stage one under the null and 
alternative hypotheses respectively. As can be seen in Table 2 the probability of stopping early 
for futility under the null is 10%.  Table 3 shows that the probability of stopping early for efficacy 
under the alternative is also 10%.  
Table 1. Stopping Boundaries  
n 
Futility Bound  
Mean Difference 
Efficacy Bound  
Mean Difference 
Futility Bound   
P-value 
Efficacy Bound  
 P-value 
~218 
-1.51088  
4.01088 
0.89986 
0.0003369 
436 
1.63584 
1.63584 
0.02493 
0.02493 
 
Table 2. Stopping probabilities given the null hypothesis 
Stage  
Futility  
Efficacy  
1  
0.10014  
0.00034  
 
Table 3. Stopping probabilities given the alternative hypothesis 
Stage  
Futility  
Efficacy  
1  
0.00034  
0.10014  
  
9.0 Risk Assessment  
This study involves randomization to one of two groups: Acetaminophen or placebo within 36 
hours of meeting inclusion. Patients will all be closely monitored in an ED or ICU setting. This 
will allow for prompt treatment of any suspected adverse events, including liver injury. Adverse 
events resulting in discontinuation of study drug will be reported to the DSMB as noted in 
section 11.1 and Appendix E.    
9.1 Potential Risks to Subjects  
Risk of Acetaminophen Administration: Serum AST and ALT will be monitored as part of the 
study on study days 0 and days 2-5.  New measured values of AST or ALT greater than or 
equal to 10 times the upper limit of normal on any measurement will prompt discontinuation of 
the study drug, which will not be restarted. Elevations in AST and ALT are common as a result 
of severe sepsis regardless of an interventional trial, are rarely associated with this dose and 
schedule of Acetaminophen, and do not require any changes in therapy other than stopping of 
the study drug. Therefore, the risk of AST or ALT elevation will be included in the informed 
consent.  AST or ALT elevations greater than 10 times the upper limit of normal will be 
considered serious adverse events.  Reporting of all adverse events to the DSMB will be done 


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in a blinded fashion. In addition to reporting these events to the DSMB, all events will also be 
included in the annual review by the IRB.    
Hypotension is uncommon but a described occurrence with intravenous Acetaminophen 
administration (frequency of 1% per manufacturer’s package insert) that appears to occur 
predominantly in patients with fever. We will monitor the need for a fluid bolus (500 ml or more), 
the need for a new vasopressor, or an increase in the vasopressor dose in the 120 minutes 
following administration of the study drug. Risk of hypotension will be included in the informed 
consent.   
Risk of Placebo Administration: Minimal risk because the volume of administration of 5% 
Dextrose in water is only 100 ml every 6 hours for 120 hours.  
Risks of Blood Draws: All patients will have blood drawn for research purposes. Many patients 
will have invasive lines placed for clinical purposes, where the risk of blood draws is extremely 
low, as blood can usually be easily obtained from these lines. In cases where an invasive line is 
not present, the risks of drawing blood are uncommon and include bleeding and bruising. 
Commonly, drawing blood is painful, and rarely, drawing blood can lead to infections at the site 
of the blood draw.  
9.2  Risk of Death  
It is possible that the treatment arm may lead to more deaths in comparison to placebo; 
mortality will be reported to the DSMB at the interim and final analyses.  
9.3  Minimization of Risk  
Federal regulations at 45 CFR 46.111(a)(1) require that risks to subjects are minimized by using 
procedures which are consistent with sound research design. There are several elements of 
study design inherent in the present protocol that meets this human subject protection 
requirement. The DSMB will be reviewing data as outlined above and will examine not only 
efficacy but safety (inclusive of mortality) and will reserve the right to halt the study at any time.   
9.4 Potential Benefits  
Study subjects may or may not receive any direct benefits from their participation in this study. It 
is unclear whether Acetaminophen will provide benefit.  Thus, the optimal treatment with 
Acetaminophen is undefined at this point.   
9.5 Risks in Relation to Anticipated Benefits  
Federal regulations at 45 CFR 46.111 (a)(2) require that “the risks to subjects are reasonable in 
relation to anticipated benefits, if any, to subjects, and the importance of the knowledge that 
may reasonably be expected to result.”  Based on the preceding assessment of risks and 
potential benefits, the risks to subjects are reasonable in relation to anticipated benefits.    


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10. Human Subjects  
Each study participant or a LAR must sign and date an informed consent form. Institutional 
review board approval will be required before any subject is entered into the study. PETAL will 
use a central IRB.  
 10.1 Selection of Subjects  
Federal regulations at 45 CFR 46(a)(3) require the equitable selection of subjects. The EDs and 
the ICUs will be screened to determine if any patient meets inclusion and exclusion criteria. 
Data that have been collected as part of the routine management of the subject will be reviewed 
to determine eligibility. No protocol-specific tests or procedures will be performed as part of the 
screening process. If any subjects meet criteria for study enrollment, then the attending 
physician will be asked for permission to approach the patient or his/her LAR for informed 
consent. Study exclusion criteria neither unjustly exclude classes of individuals from 
participation in the research nor unjustly include classes of individuals from participation in the 
research. Hence, the recruitment of subjects conforms to the principle of distributive justice.   
 10.2   Justification for Including Vulnerable Subjects  
The present research aims to investigate the safety and efficacy of a type of treatment for 
patients with sepsis induced hypotension or acute respiratory failure. Due to the nature of sepsis 
induced hypotension and respiratory failure, most of these patients will have impaired decision-
making capabilities. This study cannot be conducted if limited to enrolling only those subjects 
with retained decision-making capacity. Hence, subjects recruited for this trial are not being 
unfairly burdened with involvement in this research simply because they are easily available.  
10.3 Informed Consent  
Federal regulations 45 CFR 46.111(a)(5) require that informed consent will be sought from each 
prospective subject or the subject’s LAR. The one obtaining consent is responsible for ensuring 
that the patient and/or LAR understands the risks and benefits of participating in the study, and 
answering any questions the LAR may have throughout the study and sharing any new 
information in a timely manner that may be relevant to the LAR’s willingness to permit the 
subject’s continued participation in the trial. The consenter will make every effort to minimize 
coercion. All study participants or their LARs will be informed of the objectives of the study and 
the potential risks. The informed consent document will be used to explain the risks and benefits 
of study participation to the LAR in simple terms before the patient is entered into the study, and 
to document that the LAR is satisfied with his or her understanding of the risks and benefits of 
participating in the study and desires to participate in the study.  The investigator is responsible 
for ensuring that informed consent is given by each patient or LAR. This includes obtaining the 
appropriate signatures and dates on the informed consent document prior to the performance of 
any protocol procedures including administration of study agent.  
10.3.1 Process of Obtaining Informed Consent 
Informed consent will be obtained from the patient or from a surrogate decision maker if the 
patient lacks decision-making capacity.   In some instances, bringing a paper consent form and 
pen to the bedside of a patient with known or suspected COVID-19 and then taking these out of 
the room would violate infection control principles and policies.  Given that many of the potential 


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ASTER patients will not yet have been ruled out for COVID-19 at the time of consent, the use of 
“no-touch” consent methods will be allowed.  Additionally, the electronic consent platform 
facilitates documentation of informed consent from legal authorized representatives who are not 
present in the hospital at the time that the LAR is approached for informed consent. 
Below, we outline three examples of no-touch consent procedures that may be used: (a) a paper-
based approach (method 1); (b) a second paper-based approach (method 2); and (c) an 
electronic/e-consent approach (method 3). 
 
 Method 1 Paper-based approach:  
1. The informed consent document is delivered to the patient or LAR. 
a. If the patient or LAR is on-site, the informed consent document many be delivered to 
the patient or LAR either by research staff or by clinical staff 
b. If the LAR is off-site, the informed consent document may be emailed, faxed, or 
otherwise electronically transferred to the LAR (method dictated by institutional policy) 
2. Research staff discuss the informed consent document with the patient or LAR either in-
person or by telephone or videophone.  This step confirms subject/LAR identity. 
3. If the patient or LAR decides to consent to participate, the patient or LAR signs and dates 
the paper copy of the informed consent document. 
4. A photograph is taken of the signature page of the informed consent document and 
uploaded into the electronic database (e.g. REDCap).  
a. If using the patient’s device (such as a patient’s personal cellular phone), a survey link 
can be sent to their device to allow direct upload of the image into the electronic 
database (e.g. REDCap). 
b. If using a staff device, it must be approved to store PHI by the local institution. In that 
case, research personnel can take a photograph of the signature page of the informed 
consent document either directly or through the window or glass door leading into the 
patient’s room.  The photograph can then be uploaded into the electronic database.  If 
a staff device is taken into the patient’s room to take a photograph it must be able to 
be disinfected according to local institutional practices.  
5. Research staff (and witness if applicable based on local requirements) provide signatures 
within the electronic database (e.g. REDCap) confirming their participation in the informed 
consent process. 
6. The patient or LAR retains the paper consent document. The image of the signature page 
may be printed and bundled with a copy of the blank informed consent document for 
research records. 
 
Method 2 Paper-based approach: 
A photograph of the signed ICF can be transmitted to trial staff. 
1. An unsigned ICF is provided to the patient by a person who has entered the room. 
2. The investigator/designee arranges a telephone call or videoconference call with the patient 
(and, if desired and feasible, additional individuals requested by the patient [e.g., next of kin]). 
3. To ensure the patients are approached in a consistent fashion, a standard process should be 
used that will accomplish the following:  
a) Identification of who is on the call. 


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b) Review of the ICF with the patient by the investigator/designee and response to any 
questions the patient may have. 
c) Verbal confirmation by the patient that their questions have been answered, that they 
would like to participate in the trial, and that they have signed and dated the ICF that is in 
their possession. 
4. The patient (or an individual in the room) takes a photograph of the signed and dated ICF and 
sends it to the investigator/designee. 
5. Research staff and LAR/patient provide signatures confirming their participation in the 
informed consent process. 
6. The patient or LAR retains the paper consent document. The image of the signature page 
may be printed and bundled with a copy of the blank informed consent documents for 
research records.   
 Method 3 Electronic/e-consent approach: 
1. The electronic informed consent document is opened on a research device or a link for the 
electronic informed consent document is sent to the patient’s or LAR’s device. 
2. Research staff discuss the informed consent document with the patient or LAR either in 
person or by telephone or videophone.  This step confirms subject/LAR identity. 
3. If the patient or LAR decides to consent to participate the patient or LAR signs the electronic 
informed consent document.  This signature may be either: 
a. an actual signature (often tracing a finger on the screen) OR 
b. a username and password specific to the individual signing 
4. Research staff and witness (if required by institutional standards) provide signatures within 
the electronic database (e.g., REDCap) confirming their participation in the informed consent 
process. 
 
If a hospital device is provided to facilitate electronic or paper-based consent, that device will be 
disinfected according to institutional protocols and removed by research staff or clinical staff 
during the next entry into the patient’s room. 
This approach complies with relevant regulations and sub-regulator guidance at 45 CFR 46.117, 
45 CFR 164.512, 21 CFR 11 Subpart C (11.100–11.300), https://www.hhs.gov/ohrp/regulations-
and-policy/guidance/use-electronic-informed-consent-questions-and-answers/index.html, 
https://www.fda.gov/regulatory-information/search-fda-guidance-documents/informed-consent 
The information for the informed consent discussion will be provided in a formal document (or 
electronic equivalent) that has been approved by the IRB and in a language comprehensible to 
the potential participant, using an interpreter if necessary. The information presented in the 
consent form and by the research staff will detail the nature of the trial and what is expected of 
participants, including any potential risks or benefits of taking part. It will be clearly stated that the 
participant is free to withdraw from the trial at any time for any reason without prejudice to future 
care, and with no obligation to give the reason for withdrawal. Where a patient does not speak 
English, a short-form consent and qualified interpreter will be employed, using similar “no-touch” 
principles.  Use of an interpreter and the interpreter’s identity will be documented on the 
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 10.4   Continuing Consent  
Subjects for whom consent was initially obtained from a LAR, but who subsequently regain 
decision-making capacity while in the hospital, will be approached for consent for continuing 
participation, including continuance of data acquisition. The consent form signed by the LAR 
should reflect that such consent will be obtained. 
10.5 Withdrawal of Consent  
Patients or the LAR may withdraw or be withdrawn (by the LAR) from the trial at any time 
without prejudice. Data recorded up to the point of withdrawal will be included in the trial 
analysis, unless consent to use their data has also been explicitly withdrawn. If a patient or LAR 
requests termination of study procedures during the treatment period, the procedures will be 
stopped but the patient will continue to be followed up as part of the trial. If a patient or LAR 
withdraws consent during trial treatment, the study procedures will be stopped but permission 
will be sought to access medical records for data related to the trial. If a patient or LAR wishes 
to withdraw from the trial after completion of trial treatment, permission to access medical 
records for trial data will be assumed unless explicitly withdrawn.    
 10.6    Identification of Legally Authorized Representatives  
Many of the patients approached for participation in this research protocol will have limitations of 
decision-making abilities due to their critical illness. Hence, some patients will not be able to 
provide informed consent.  Accordingly, informed consent will be sought from the potential 
subject’s legally authorized representative.    
Regarding proxy consent, the existing federal research regulations (‘the Common Rule’) states 
at 45 CFR 46.116 that “no investigator may involve a human being as a subject in 
research…unless the investigator has obtained the legally effective informed consent of the 
subject or the subject’s legally authorized representative”; and defines at 45 CFR 46 102 (c) a 
legally authorized representative as “an individual or judicial or other body authorized under 
applicable law to consent on behalf of a prospective subject to the subject’s participation in the 
procedures involved in the research.” The Office of Human Research Protections (OHRP) 
defined examples of “applicable law” as being state statutes, regulations, case law, or formal 
opinion of a State Attorney General that addresses the issue of surrogate consent to medical 
procedures. Such “applicable law” could then be considered as empowering the LAR to provide 
consent for subject participation in the research. Interpretation of “applicable law” may be state 
specific and will be addressed by the PETAL central IRB.  
According to a previous President’s Bioethics Committee (National Bioethics Advisory 
Committee (NBAC)), an investigator should accept a relative or friend of the potential subject 
who is recognized as an LAR for purposes of clinical decision making under the law of the state 
where the research takes place [44]. Finally, OHRP has stated in their determination letters that 
a surrogate could serve as a LAR for research decision making if such an individual is 
authorized under applicable state law to provide consent for the “procedures” involved in the 
research study [44].  


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10.7    Justification of Surrogate Consent  
According to the Belmont Report, respect for persons incorporates at least two ethical 
convictions; first, that individuals should be treated as autonomous agents, and second, that 
person with diminished autonomy are entitled to protection. One method that serves to protect 
subjects is restrictions on the participation of subjects in research that presents greater than 
minimal risks. Commentators and research ethics commissions have held the view that it is 
permissible to include incapable subjects in greater than minimal risk research as long as there 
is the potential for beneficial effects and that the research presents a balance of risks and 
expected direct benefits similar to that available in the clinical setting [45]. Several U.S. task 
forces have deemed it is permissible to include incapable subjects in research. For example, the 
American College of Physicians’ document allows surrogates to consent to research involving 
incapable subjects only  “if the net additional risks of participation are not substantially greater 
than the risks of standard treatment.” [46]. Finally, NBAC stated that an IRB may approve a 
protocol that presents greater than minimal risk but offers the prospect of direct medical benefits 
to the subject, provided that “the potential subject’s LAR gives permission…” [44]. Consistent 
with the above ethical sensibilities regarding the participation of decisionally incapable subjects 
in research and the previous assessment of risks and benefits in the previous section, the 
present trial presents a balance of risks and potential direct benefits that is similar to that 
available in the clinical setting, with the exception of the additional blood draws.      
 10.8     Additional Safeguards for Vulnerable Subjects  
The present research will involve subjects who might be vulnerable to coercion or undue 
influence. As required in 45CFR46.111 (b), we recommend that sites utilize additional 
safeguards to protect the rights and welfare of these subjects. Such safeguards might include 
but are not limited to: a) assessment of the potential subject’s capacity to provide informed 
consent, b) the availability of the LAR to monitor the subject’s subsequent participation and 
withdrawal from the study; c) augmented consent processes. The specific nature of the 
additional safeguards will be left to the discretion of the central IRB, in conjunction with the sites.    
 10.9     Confidentiality  
Federal regulations at 45 CFR 46 111 (a) (7) requires that when appropriate, there are 
adequate provisions to protect the privacy of subjects and to maintain the confidentiality of data. 
To maintain confidentiality, all laboratory specimens, evaluation forms, and reports will be 
identified only by a coded number.  The coded number will be generated by a computer, and 
only the study team will have access to the codes.  All records will be kept in a locked, 
password protected computer.  All computer entry and networking programs will be done with 
coded numbers only.  All paper case report forms will be maintained inside a locked office.  
Study information will not be released without the written permission of the patient, except as 
necessary for monitoring by the National Heart, Lung, and Blood Institute, and the PETAL 
Clinical Coordinating Center.  


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11. Adverse Events  
11.1 Safety Monitoring  
Assuring patient safety is an essential component of this protocol. Each participating 
investigator has primary responsibility for the safety of the individual participants under his 
or her care. The Investigators will determine daily if any adverse events occur during the 
period from enrollment through study day 10 (five days after completion of the study drug 
infusions) or hospital discharge, whichever occurs first and will determine if such adverse 
events are reportable.   
The following adverse events will be considered reportable and thus collected in the adverse 
event case report forms:  
  
 
Serious adverse events  
 
Non-serious adverse events that are considered by the investigator to be related to 
study procedures or of uncertain relationship (Appendix E)  
 
Study-specific clinical outcomes (Primary and Secondary Outcomes and 
Assessments During the Study), including serious adverse events such as organ 
failures and death, are systematically recorded in the case report forms and are 
exempt from adverse event reporting unless the investigator deems the event to 
be related to the administration of study drug or the conduct of study procedures 
(or of uncertain relationship) as outlined in Appendix E.   
After randomization, adverse events must be evaluated by the investigator. If the adverse event 
is judged to be reportable, as outlined above, then the investigator will report to the CCC their 
assessment of the potential relatedness of each adverse event to the study drug or protocol 
procedure via electronic data entry. Investigators will assess if there is a reasonable possibility 
that the study procedure caused the event, based on the criteria outlined in Appendix E. 
Investigators will also consider if the event is unexpected.  Unexpected adverse events are 
events not listed in the protocol and the investigator brochure for intravenous Acetaminophen. 
Investigators will also determine if adverse events are unanticipated given the patient’s clinical 
course, previous medical conditions, and concomitant medications.    
If a patient's treatment is discontinued as a result of an adverse event, study site personnel 
must report the circumstances and data leading to discontinuation of treatment in the adverse 
event case report forms.    
11.2 Serious Adverse Events  
Serious adverse event collection begins after randomization and study procedures have been 
initiated.  If a patient experiences a serious adverse event after consent, but prior to 
randomization or starting study procedures, the event will NOT be collected. Study site 
personnel must alert the CCC of any serious and study procedure related adverse event 
within 24 hours of investigator awareness of the event.  Alerts issued via telephone are to be 
immediately followed with official notification on the adverse event case report form. See 
Appendix E for reporting timelines for serious, unexpected, study related events (SAEs) and 
serious, unexpected suspected adverse reactions (SUSARs).  


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As per the FDA and NIH definitions, a serious adverse event is any adverse event that results in 
one of the following outcomes:  
 Death   
 A life-threatening experience (that is, immediate risk of dying)  
 Prolonged inpatient hospitalization or re-hospitalization  
As per http://www.fda.gov/Safety/MedWatch/HowToReport/ucm053087.htm: Report if 
admission to the hospital or prolongation of hospitalization was a result of the adverse 
event. Emergency room visits that do not result in admission to the hospital should be 
evaluated for one of the other serious outcomes (e.g., life-threatening; required 
intervention to prevent permanent impairment or damage; other serious medically 
important event).  
 Persistent or significant disability/incapacity  
As per http://www.fda.gov/Safety/MedWatch/HowToReport/ucm053087.htm: Report if the 
adverse event resulted in a substantial disruption of a person's ability to conduct normal 
life functions, i.e., the adverse event resulted in a significant, persistent or permanent 
change, impairment, damage or disruption in the patient's body function/structure, physical 
activities and/or quality of life.  
Reportable serious adverse events that may not result in death, be life-threatening, or require 
hospitalization may be considered serious adverse events when, based upon appropriate 
medical judgment, they may jeopardize the patient and may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition.  
Serious adverse events will be collected during the first 10 study days or until hospital  
discharge, whichever occurs first, regardless of the investigator's opinion of causation.     
 
 


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Appendices  
A. Schedule of Events   
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Creatinine: 
• 
Prehospital: Most recent outpatient creatinine from 24 hours to 365 days prior to admission if available  
• 
Baseline: Lowest in hospital creatinine prior to randomization 
• 
On study: Collect peak creatinine between randomization and study day 28 and last creatinine value prior to 
death, RRT, discharge from study hospital, or day 28 (whichever comes first) 
**Liver Function Tests;: These values are required on days 2-5, and 7 (day 7 can be collected ± 1 day).   
aDay-2 Plasma: Collect 30 minutes prior to study drug administration; subject must have received at least 5 doses 
of study drug before the day 2 samples are obtained).  One aliquot of Day 2 collection will be designated for 
Acetaminophen trough level. 
***First dose of study drug should be administered within 4 hours of randomization (Day 0). In the 120 minute interval 
following all study drug doses, instances of the administration of fluid boluses, new vasopressors, and increases in 
vasopressor doses will be recorded including maximum fluid volume, vasopressor dose, or greatest increase in 
vasopressor dose. 


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B. Sequential (Sepsis-related) Organ Failure Assessment (SOFA) Score   
SOFA Score  
0  
1  
2  
3  
4  
Pulmonary (PaO2/FiO2, 
mmHg)  
≥ 400  
< 400  
< 300  
< 200  
< 100  
Coagulation (Platelets x 
103/mm3)  
≥ 150  
< 150  
< 100  
< 50  
< 20  
Liver (Total Bilirubin, 
mg/dL)  
< 1.2  
1.2 – 1.9  
2.0 – 5.9  
6.0 – 11.9  
≥ 12.0  
Cardiovascular (Mean 
Arterial Pressure, 
mmHg)  
≥ 70  
< 70  
Dopamine ≤ 5  
or dobutamine   
(any dose)*  
Dopamine > 5 or 
epinephrine ≤ 0.1 
or norepinephrine 
≤ 0.1*  
Dopamine > 15 or 
epinephrine > 0.1 or 
norepinephrine > 
0.1*  
Renal (Serum 
Creatinine, mg/dL)  
< 1.2  
1.2 – 1.9  
2.0 – 3.4  
3.5 – 4.9  
≥ 5.0  
*Adrenergic agents administered for at least 1 hour, doses given as μg/kg/min  
  


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C. Ventilator Management for Patients with ARDS  
PART I: VENTILATOR SETUP AND ADJUSTMENT  
1. Calculate predicted body weight (PBW) in kilograms  
Males = 50 + 2.3 [height (inches) - 60]  
Females = 45.5 + 2.3 [height (inches) -60]  
2. Controlled ventilation mode required during NMB.  After NMB, select any ventilator mode 
capable of delivering the prescribed VT (6ml/kg PBW, +/- 2 ml/kg)   
  
 
OXYGENATION GOAL: PaO2 55-80 mmHg or SpO2 88-95%  
1. Use a minimum PEEP of 5 cm H2O.   
2. Adjust FiO2 or PEEP upward within 5 minutes of consistent measurements below the 
oxygenation target range  
3. Adjust FiO2 or PEEP downward within 30 minutes of consistent measurements above the 
oxygenation target range.    
4. The below PEEP strategy FiO2/PEEP table should be used in all patients.  
 
PEEP/FiO2  
FiO2  
.30  .40  .40  
.50  
.60  .70  
.80  .90  
1.0  
PEEP  
5  
5  
 8  
8-10  10  10-14  14  14-18  
18-24  
 
PLATEAU PRESSURE GOAL: ≤ 30 cm H2O  
Check Pplat (0.5 second inspiratory pause), at least q 4h and after each change in PEEP or VT.  
• 
If Pplat > 30 cm H2O: decrease VT by 1ml/kg steps (minimum = 4 ml/kg).  
• 
If Pplat < 25 cm H2O and VT< 6 ml/kg, increase VT by 1 ml/kg until Pplat > 25 cm H2O 
or VT = 6 ml/kg.  
• 
If “severe dyspnea” (more than 3 double breaths per minute or airway pressure 
remains at or below PEEP level during inspiration), then raise VT to 7 or 8 ml/kg PBW if 
Pplat remains below 30.  If Pplat exceeds 30 cm H20, then revert to lower VT and 
consider more sedation.  
pH GOAL: ≥ 7.30  
• 
If pH < 7.30: May give NaHCO3 (neither encouraged nor discouraged)  
• 
If pH < 7.15: VT may be raised and Pplat limit suspended (neither encouraged nor 
discouraged)  
• 
If pH < 7.30: incremental increase in RR allowed to max rate of 35.  


ASTER Protocol Version 4.4 
July 7, 2022 
 
 
 
 
 
 
 
 
 
 
Pg.  49     
  
  
I:E RATIO GOAL: Recommend that duration of inspiration be ≤ duration of expiration.  
PART II: WEANING  
Conduct a SPONTANEOUS BREATHING TRIAL daily when:  
• 
FiO2 ≤ 0.40 and PEEP ≤ 8 
• 
PEEP and FiO2 ≤ values of previous day 
• 
Systolic BP ≥ 90 mmHg and without vasopressor support 
• 
Spontaneous respirations  
SPONTANEOUS BREATHING TRIAL (SBT):  
If all above criteria are met initiate a trial of UP TO 120 minutes of spontaneous breathing with 
FiO2 < 0.5 using any of the following approaches:  
• 
Pressure support < 5cm H2O, PEEP < 5cm H2O  
• 
CPAP < 5 cm H2O   
• 
T-piece  
• 
Tracheostomy mask   
Assess for tolerance using the following:  
• 
SpO2 ≥ 90: and/or PaO2 ≥ 60 mmHg  
• 
Spontaneous VT ≥ 4 ml/kg PBW  
• 
RR ≤ 35/min  
• 
pH ≥ 7.3  
• 
No respiratory distress (distress = 2 or more of the following):  
o 
HR > 120% of baseline 
o 
Marked accessory muscle use  
o 
Abdominal paradox  
o 
Diaphoresis 
o 
Marked dyspnea  
If tolerated for at least 30 minutes, consider extubation.  
If not tolerated resume pre-weaning settings.  
  
  
 
 


ASTER Protocol Version 4.4 
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D. On-Study Fluid Management for Patients with ARDS  
This protocol should be initiated after the initial resuscitation.  
o Discontinue maintenance fluids.  
o Continue medications and nutrition.  
o Manage electrolytes and blood products per usual practice.  
o For shock, use any combination of fluid boluses# and vasopressor(s) to achieve MAP ≥ 
60 mmHg as fast as possible.  Wean vasopressors as quickly as tolerated beginning four 
hours after blood pressure has stabilized.  
o Withhold diuretic therapy in renal failure § and until 12 hours after last fluid bolus or 
vasopressor given.  
  
  
CVP  
(recommend)  
  
  
  
PAOP  
(optional)  
  
MAP > 60 mm Hg AND off vasopressors for > 12 hours  
Average urine output < 0.5 
ml/kg/hr  
Average urine output > 0.5 
ml/kg/hr  
>8  
> 12  
Furosemide* 
Reassess in 1 hour  
Furosemide* 
Reassess in 4 hours  
4-8  
8-12  
Give fluid bolus as fast as 
possible# Reassess in 1 hour  
  
< 4  
< 8  
No intervention 
Reassess in 4 hours  
  
  
§ Renal failure is defined as dialysis dependence, oliguria with serum creatinine > 3mg/dl, or oliguria with serum 
creatinine 0-3 with urinary indices indicative of acute renal failure as in the original FACTT trial.  
# Recommended fluid bolus= 15 mL / kg crystalloid (round to nearest 250 mL) or 1 Unit packed red cells or 25 
grams albumin  
*Recommended Furosemide dosing = begin with 20 mg bolus or 3 mg / hr infusion or last known effective dose.  
Double each subsequent dose until goal achieved (oliguria reversal or intravascular pressure target) or maximum 
infusion rate of 24 mg / hr or 160 mg bolus reached.  Do not exceed 620 mg / day.  Also, if patient has heart failure, 
consider treatment with dobutamine.  
  
NOTE: For patients without a central venous catheter, no fluid gain over the first 7 study days is 
recommended once patients’ blood pressure has stabilized.  Stable blood pressure is defined as 
no requirement for either vasopressors or a fluid bolus to support blood pressure for 12 or more 
hours.   
  
 
 


ASTER Protocol Version 4.4 
July 7, 2022 
 
 
 
 
 
 
 
 
 
 
Pg.  51     
E. Adverse Event Reporting and Unanticipated Events  
As noted in section 11, investigators will report all adverse events that are serious and study 
drug or study procedure related (or of uncertain relatedness) to the CCC within 24 hours. The 
CCC will then notify the NHLBI and Central Institutional Review Board (cIRB).   
The Medical Monitor at the CCC will work collaboratively with the reporting investigator to 
determine if a serious adverse event has a reasonable possibility of having been caused by the 
study drug or study procedure, as outlined in 21 CFR 312.3 (1), and below. The Medical Monitor 
will be unblinded and will also determine if the event is unexpected for Acetaminophen. An 
adverse event is considered “unexpected” if it is not listed in the investigator brochure or the 
study protocol (21 CFR 312.32(a)). If a determination is made that a serious adverse event has a 
reasonable possibility of having been caused by a study procedure or the study drug, it will be 
classified as a suspected adverse reaction. If the suspected adverse reaction is unexpected, it 
will be classified as a serious unexpected suspected adverse reaction (SUSAR).   
The CCC will report all unexpected deaths, serious and treatment related adverse events, and 
SUSARs to the DSMB, NHLBI, and cIRB within 7 days after receipt of the report from a clinical 
site. A written report will be sent to the NHLBI, DSMB and the cIRB within 15 calendar days. The 
DSMB will also review all adverse events and clinical outcomes during scheduled interim 
analyses. The CCC will distribute the written summary of the DSMB’s periodic review of adverse 
events to the cIRB in accordance with NIH guidelines 
(http://grants.nih.gov/grants/guide/noticefiles/not99-107.html).   
E.1. Unanticipated Problems (UP) 
Investigators must also report Unanticipated Problems, regardless of severity, associated with 
study procedures within 24 hours. An unanticipated problem is defined as follows: any incident, 
experience, or outcome that meets all of the following criteria:   
• 
Unexpected, in terms of nature, severity, or frequency, given the research procedures that 
are described in the protocol-related documents, such as the IRB-approved research 
protocol and informed consent document; and the characteristics of the subject population 
being studied;   
• 
Related or possibly related to participation in the research. Possibly related means there is a 
reasonable possibility that the incident, experience, or outcome may have been caused by 
the procedures involved in the research;   
• 
Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized.   
E.2. Determining Relationship of Adverse Events to Procedures  
Investigators will be asked to grade the strength of the relationship of an adverse event to study 
procedures as follows:   
• 
Definitely Related: The event follows: a) A reasonable, temporal sequence from a study 
procedure; and b) Cannot be explained by the known characteristics of the patient’s 
clinical state or other therapies; and c) Evaluation of the patient’s clinical state indicates 
to the investigator that the experience is definitely related to study procedures.   


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• 
Probably or Possibly Related: The event should be assessed following the same criteria 
for “Definitely Related”. If in the investigator’s opinion at least one or more of the criteria 
are not present, then “probably” or “possibly” associated should be selected.   
• 
Probably Not Related: The event occurred while the patient was on the study but can 
reasonably be explained by the known characteristics of the patient’s clinical state or 
other therapies.   
• 
Definitely Not Related: The event is definitely produced by the patient’s clinical state or by 
other modes of therapy administered to the patient.   
• 
Uncertain Relationship: The event does not meet any of the criteria previously outlined.   
E.3. Clinical Outcomes that may be Exempt from Adverse Event Reporting  
Study-specific clinical outcomes of sepsis, as outlined in Section 8.1 and 8.3 (Primary and 
Secondary Outcomes) and Section 7.2 (Assessments During the Study) are exempt from 
adverse event reporting unless the investigator deems the event to be related to the study 
procedures (or of uncertain relationship) or if the event leads to discontinuation of study 
procedures. The following are examples of events that will be considered study specific clinical 
outcomes:   
• 
Death not related to the study procedures   
• 
Cardiovascular events: need for vasoactive drugs or fluids for hypotension or 
hypotension not temporally related to study drug infusion  
• 
Respiratory events: decreased PaO2/FiO2, hypoxia, worsening acute respiratory 
distress syndrome, or respiratory failure.   
• 
Hepatic events: hepatic injury or liver dysfunction that leads to an increase from baseline 
in the serum level of bilirubin.   
• 
Renal events: renal failure, renal insufficiency, or renal injury that leads to an increase 
from baseline in serum creatinine.   
 
 


ASTER Protocol Version 4.4 
July 7, 2022 
 
 
 
 
 
 
 
 
 
 
Pg.  53     
F. Alcohol Use Screen Audit C  
1. How often do you have a drink containing alcohol?  
 Never (0 points)  
 Monthly or less (1 point)  
 2-4 times a month (2 points)  
 2-3 times a week (3 points)  
 4 or more times a week (4 points)  
2. How many standard drinks containing alcohol do you have on a typical day?  
  1 or 2 (0 points)  
  3 or 4 (1 point)  
  5 or 6 (2 points)  
  7 or 9 (3 points)  
  10 or more (4 points)  
3. How often do you have six or more drinks on one occasion?  
  Never (0 points)  
  Less than monthly (1 point)  
  Monthly (2 points)  
  Weekly (3 points)  
  Daily or almost daily (4 points)  
TOTAL POINTS:  ______  
If total > 4, patient is excluded from the study.  
   
  
  


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July 7, 2022 
 
 
 
 
 
 
 
 
 
 
Pg.  54     
G. Fever Management Guidelines   
Use of nonsteroidal anti-inflammatory drugs (NSAIDs) or Cyclooxygenase-2 inhibitors to treat 
fever will be strongly discouraged during the 5 day study drug administration and the use of 
these medications for other indications will also be discouraged. No restriction will be placed on 
the use of aspirin in doses of less than or equal to 325 mg per day. The use of open-label 
acetaminophen will not be permitted while receiving study medication. Once the course of study 
drug administration has been completed, open-label acetaminophen can be administered at the 
discretion of the treating clinician. Physical cooling will be used at the discretion of the clinical 
team per local practice preferences (modified from Reference 29).   
   
  
 


ASTER Protocol Version 4.4 
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Pg.  55     
References   
1. 
Lamontagne, F., et al., Intravenous Vitamin C in Adults with Sepsis in the Intensive Care 
Unit. N Engl J Med, 2022. 
2. 
Angus, D.C., et al., Epidemiology of severe sepsis in the United States:  analysis of 
incidence, outcome, and associated costs of care. Crit Care Med, 2001. 29: p. 1303-1310.  
3. 
Martin, G.S., et al., The epidemiology of sepsis in the United States from 1979 through 
2000. N Engl J Med, 2003. 348: p. 1546-1554.  
4. 
Bellani, G., et al., Epidemiology, Patterns of Care, and Mortality for Patients With Acute 
Respiratory Distress Syndrome in Intensive Care Units in 50 Countries. JAMA, 2016. 315(8): p. 
788-800.  
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Stapleton, R.D., et al., Causes and timing of death in patients with ARDS. Chest, 2005. 
128(2): p. 525-32.  
6. 
Linder, A., et al., Small acute increases in serum creatinine are associated with 
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1075-81.  
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Chertow, G.M., et al., Acute kidney injury, mortality, length of stay, and costs in 
hospitalized patients. J Am Soc Nephrol, 2005. 16(11): p. 3365-70.  
8. 
Rewa, O. and S.M. Bagshaw, Acute kidney injury-epidemiology, outcomes and 
economics. Nat Rev Nephrol, 2014. 10(4): p. 193-207.  
9. 
Liu, K.D., et al., Predictive and pathogenetic value of plasma biomarkers for acute kidney 
injury in patients with acute lung injury. Crit Care Med, 2007. 35(12): p. 2755-61.  
10. 
Gotts, J.E. and M.A. Matthay, Sepsis: pathophysiology and clinical management. BMJ, 
2016. 353: p. i1585.  
11. 
Piagnerelli, M., et al., Red blood cell rheology in sepsis. Intensive Care Med, 2003. 29(7): 
p. 1052-61.  
12. 
Machiedo, G.W., et al., The incidence of decreased red blood cell deformability in sepsis 
and the association with oxygen free radical damage and multiple-system organ failure. Arch 
Surg, 1989. 124(12): p. 1386-9.  
13. 
Baskurt, O.K., D. Gelmont, and H.J. Meiselman, Red blood cell deformability in sepsis. 
Am J Respir Crit Care Med, 1998. 157(2): p. 421-7.  
14. 
Larsen, R., et al., A central role for free heme in the pathogenesis of severe sepsis. Sci 
Transl Med, 2010. 2(51): p. 51ra71.  
15. 
Janz, D.R., et al., Association Between Cell-Free Hemoglobin, Acetaminophen, and 
Mortality in Patients With Sepsis: An Observational Study. Crit Care Med, 2013.  
16. 
Adamzik, M., et al., Free hemoglobin concentration in severe sepsis: methods of 
measurement and prediction of outcome. Crit Care, 2012. 16(4): p. R125.  
17. 
Reiter, C.D., et al., Cell-free hemoglobin limits nitric oxide bioavailability in sicklecell 
disease. Nat Med, 2002. 8(12): p. 1383-9.  
18. 
Meyer, C., et al., Hemodialysis-induced release of hemoglobin limits nitric oxide 
bioavailability and impairs vascular function. J Am Coll Cardiol, 2010. 55(5): p. 454-9.  


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Pg.  56     
19. 
Gladwin, M.T. and D.B. Kim-Shapiro, Storage lesion in banked blood due to hemolysis-
dependent disruption of nitric oxide homeostasis. Curr Opin Hematol, 2009. 16(6): p. 515-23.  
20. 
Natanson, C., et al., Cell-free hemoglobin-based blood substitutes and risk of myocardial 
infarction and death: a meta-analysis. JAMA, 2008. 299(19): p. 230412.  
21. 
Lin, T., et al., Synergistic inflammation is induced by blood degradation products with 
microbial Toll-like receptor agonists and is blocked by hemopexin. J Infect Dis, 2010. 202(4): p. 
624-32.  
22. 
Dutra, F.F., et al., Hemolysis-induced lethality involves inflammasome activation by 
heme. Proc Natl Acad Sci U S A, 2014. 111(39): p. E4110-8.  
23. 
Dutra, F.F. and M.T. Bozza, Heme on innate immunity and inflammation. Front 
Pharmacol, 2014. 5: p. 115.  
24. 
Billings, F.T.t., et al., Postoperative acute kidney injury is associated with 
hemoglobinemia and an enhanced oxidative stress response. Free Radic Biol Med, 2011. 
50(11): p. 1480-7.  
25. 
Reeder, B.J., et al., The radical and redox chemistry of myoglobin and hemoglobin: from 
in vitro studies to human pathology. Antioxid Redox Signal, 2004. 6(6): p. 954-66.  
26. 
Ware, L.B., et al., Plasma biomarkers of oxidant stress and development of organ failure 
in severe sepsis. Shock, 2011. 36(1): p. 12-7.  
27. 
Boutaud, O., et al., Acetaminophen inhibits hemoprotein-catalyzed lipid peroxidation and 
attenuates rhabdomyolysis-induced renal failure. Proc Natl Acad Sci U S A, 2010. 107(6): p. 
2699-704.  
28. 
Ouellet, M. and M.D. Percival, Mechanism of acetaminophen inhibition of cyclooxygenase 
isoforms. Arch Biochem Biophys, 2001. 387(2): p. 273-80.  
29. 
Anderson, B.J., Paracetamol (Acetaminophen): mechanisms of action. Paediatr Anaesth, 
2008. 18(10): p. 915-21.  
30. 
Janz, D.R., et al., Randomized, placebo-controlled trial of acetaminophen for the 
reduction of oxidative injury in severe sepsis: the Acetaminophen for the Reduction of Oxidative 
Injury in Severe Sepsis trial. Crit Care Med, 2015. 43(3): p. 534-41.  
31. 
Plewes, K.A., et al., Paracetamol as adjunctive treatment in severe and moderately 
severe falciparum malaria: An open label, randomised controlled trial. 2015: Presented at 
American Society of Tropical Medicine and Hygiene, October p. Abstract LB-5377.  
32. 
Young, P., et al., Acetaminophen for Fever in Critically Ill Patients with Suspected 
Infection. N Engl J Med, 2015. 373(23): p. 2215-24.  
33. 
Watkins, P.B., et al., Aminotransferase elevations in healthy adults receiving 4 grams of 
acetaminophen daily: a randomized controlled trial. JAMA, 2006. 296(1): p. 87-93.  
34. 
Heard, K., et al., A randomized, placebo-controlled trial to determine the course of 
aminotransferase elevation during prolonged acetaminophen administration. BMC Pharmacol 
Toxicol, 2014. 15: p. 39.  
35. 
Heard, S.O., et al., The influence of lipsome-encapsulated prostaglandin E1 on hydrogen 
peroxide concentrations in the exhaled breath of patients with the acute respiratory distress 
syndrome. Anesth Analg, 1999. 89: p. 353-357.  


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Pg.  57     
36. 
McNicol, E.D., et al., Single-dose intravenous paracetamol or propacetamol for 
prevention or treatment of postoperative pain: a systematic review and metaanalysis. Br J 
Anaesth, 2011. 106(6): p. 764-75.  
37. 
Jahr, J.S. and V.K. Lee, Intravenous acetaminophen. Anesthesiol Clin, 2010. 28(4): p. 
619-45.  
38. 
Rice, T.W., et al., Randomized trial of initial trophic versus full-energy enteral nutrition in 
mechanically ventilated patients with acute respiratory failure. Crit Care Med, 2011. 39(5): p. 
967-74.  
39. 
Russell, J.A., et al., Days alive and free as an alternative to a mortality outcome in pivotal 
vasopressor and septic shock trials. J Crit Care, 2018. 47: p. 333-337.  
40. 
Geven, C., et al., Preclinical safety evaluation of the adrenomedullin-binding antibody 
Adrecizumab in rodents, dogs and non-human primates. Toxicol Appl Pharmacol, 2019. 369: p. 
1-16.  
41. 
Kahn, S.A. and C.W. Lentz, Fictitious hyperglycemia: point-of-care glucose measurement 
is inaccurate during high-dose vitamin C infusion for burn shock resuscitation. J Burn Care Res, 
2015. 36(2): p. e67-71.  
42. 
Haybittle, J.L., Repeated assessment of results in clinical trials of cancer treatment. Br J 
Radiol, 1971. 44(526): p. 793-7.  
43. 
Peto, R., et al., Design and analysis of randomized clinical trials requiring prolonged 
observation of each patient. I. Introduction and design. Br J Cancer, 1976. 34(6): p. 585-612.  
44. 
National Bioethics Advisory Commitee (NBAC), Research Involving Persons with Mental 
Disorders That May Affect Decisionmaking Capacity. 1998, Rockville: U.S. Government Printing 
Office.  
45. 
Dresser, R., Research Involving Persons with Mental Disabilities: A Review of Policy 
Issues and Proposals., in National Bioethics Advisory Commission. 1999, U.S. Government 
Printing Office: Rockville. p. 5-28.  
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American College of Physicians, Cognitively Impaired Subjects. Ann Int Med, 1989. 111: 
p. 843-8.  
 

ICF_003.pdf: 
PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  06/21/2021 
  
 
1 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
Part 1 of 2: MASTER CONSENT 
 
Name of participant: __________________________________________________________ Age: ___________ 
 
You are being invited to take part in a research study. This form will tell you about this study. Please read this 
form carefully and ask any questions you may have. This study is a multi-site study, meaning it will take place 
at several different places. Because this is a multi-site study this consent form includes two parts. Part 1 of this 
consent form is the Master Consent and includes information that applies to all study sites. Part 2 of the 
consent form is the Study Site Information and includes information specific to the study site where you are 
being asked to participate. Both parts together are the legal consent form and must be provided to you. 
 
Key information about this study: 
You are being asked to take part in this study because you have a severe infection (sepsis). In this study we are 
trying to understand whether acetaminophen (Tylenol) and/or vitamin C (ascorbate) helps patients recover from 
sepsis compared to patients treated with a placebo (sugar water).  Sepsis causes damage to red blood cells 
which can release a protein into the blood called hemoglobin.  Hemoglobin can cause injury to the blood vessels 
and organs of patients with sepsis. In laboratory studies and in small studies of patients with sepsis, 
acetaminophen or vitamin C can stop some of the effects of hemoglobin. This may then limit injury to blood 
vessels and organs, and help people recover from sepsis.  
Both drugs have been used many times in humans with good safety. Possible risks of acetaminophen include 
liver injury or lower blood pressure. Possible risks of vitamin C are kidney stones and change of blood sugar 
tests.  You will be carefully monitored for these if you choose to participate.  
If you agree to be in this study you could receive Tylenol, vitamin C, or a placebo through an IV for up to 5 days 
while you are in the ICU. We will collect information from your medical record while you are in the hospital.  
Acetaminophen or vitamin C are not given to all sepsis patients outside of research, and if you do not join, your 
doctor will choose all therapies. 
You may choose not to be in this study. If you choose not to be in the study, you will receive the medical 
treatment that you are receiving now and anything else that your doctors think might be helpful.  You can stop 
being in this study at any time.  
If we learn something while doing this study that could change the risks or benefits of you being in it, we will tell 
you so that you can decide if you want to stay in or join this study.  
Key Information:  
The first section of this document contains some key points that the research team thought you 
would find important. The study is described in more detail after this section. If you do not 
understand something, please ask someone.  
Date of IRB Approval: 07/14/2021
Date of Expiration: 07/13/2022


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  06/21/2021 
  
 
2 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
The word “you” in this form refers to the person who will be in the study. Your legal representative will be asked 
to read and sign this consent form to give permission for you to participate if you are unable to do so yourself.    
 
 
What is the purpose of this study?  
We are studying if vitamin C or acetaminophen helps people with sepsis. The study is paid for by the National 
Institutes of Health.  
Sepsis can lower your blood pressure and worsen the function of your internal organs, including the lungs and 
kidneys. These effects of sepsis can be reversible (can be fixed). We are studying whether vitamin C or 
acetaminophen improves your blood pressure and/or the function of your lungs and kidneys.  
If you agree to join the study, you will be assigned by chance (like a coin toss) to one of four groups. One group 
gets vitamin C intravenously (through a small tube in a vein) for 5 days. One group gets acetaminophen 
intravenously for 5 days. The final two groups get a placebo (simple sugar water that does not contain vitamin C 
or acetaminophen) for 5 days. All groups will receive standard treatment for sepsis, including antibiotics, as 
directed by your treating physicians.  
What will happen and how long will you be in the study? 
 Overview: We seek 900 people with sepsis to join this study in about 50 hospitals. Your hospital is one of 
these hospitals and expects to enroll 30 or more people.  If you choose to participate in this study, you will 
be in the study for three months (90 days).   
 Review of your medical information: First, we reviewed your medical information to ensure you are eligible 
to join the research study.  
 Pregnancy Check: If you agree to join this study and  are a woman who can have children, we will do a 
pregnancy test before you join the study, if your doctor has not already checked this as part of your normal 
care. If you are pregnant, you may not join the study. 
 Laboratory Tests: We will check your liver function levels in your blood before you join the study, if your 
doctor has not already checked this as part of your normal care. If you already have very high liver tests you 
will not be allowed to join the study.  
 
 
Detailed Information: 
The rest of this document includes detailed information about this study (in addition to the 
information listed above). 
Date of IRB Approval: 07/14/2021
Date of Expiration: 07/13/2022


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  06/21/2021 
  
 
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CIRB USE ONLY 
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If you are eligible to be in the study, and you agree to participate:  
 
 
Side effects and risks that you can expect if you take part in this study:  
Vitamin C: No serious or harmful side effects of intravenous vitamin C have been seen in previous studies. 
Potential risks include: dry mouth, nausea, vomiting, dizziness, and headache. Very rarely and usually without 
warning, an allergic reaction is possible; of these, most are mild and very few are life threatening. If you develop 
an allergic reaction to vitamin C, your doctors will determine if treatment is needed. There is also a rare chance 
that you may develop a kidney stone from vitamin C therapy. 
At high doses, administration of vitamin C makes blood glucose (blood sugar) tests that are done by a fingerstick 
at the bedside not reliable. If you are in the Vitamin C-ACTIVE or Vitamin C-Placebo group, when your doctors 
want blood glucose tests, your blood will be sent to the clinical laboratory at your hospital, which has a different 
machine to measure blood glucose. Since most blood glucose testing in the ICU is usually done using an in-room 
testing machine, using the clinical laboratory at your hospital could be different than standard practice. It may 
take up to 60 minutes to get results, which may delay the treatment of high or low blood glucose.  
Acetaminophen: At high doses, acetaminophen can injure the liver, especially in patients who already have 
liver problems. For your safety, if you are in the Acetaminophen-ACTIVE or Acetaminophen-Placebo group, we 
will monitor your liver tests and will stop the study medicine if your liver tests suggest that your liver function is 
worsening. Stopping acetaminophen usually results in liver tests returning to normal without permanent 
damage.  
Date of IRB Approval: 07/14/2021
Date of Expiration: 07/13/2022


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  06/21/2021 
  
 
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CIRB USE ONLY 
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Less commonly, acetaminophen can lower blood pressure. If you are in the Acetaminophen-ACTIVE or 
Acetaminophen-Placebo group, we will watch your blood pressure after each dose of the study medicine. If your 
blood pressure goes down too much, your doctors may treat the low blood pressure as part of regular clinical 
care.  
An allergic reaction is possible but rare; most are mild, with very few allergic reactions being life threatening. If 
you develop an allergic reaction to acetaminophen, your doctors will treat it.   
Drawing Blood: There are no major risks associated with drawing blood. We will usually take blood from tubes 
that are already in your vein or artery. If you do not have a tube in a vein or artery, then we will get the blood 
with a needle. You may experience minor discomfort, bruising or soreness from the needle. Drawing blood 
through a needle very rarely causes infection. Blood draws will be done by a trained professional.  
Health Information Privacy: To ensure your privacy, your health information will be handled securely by the 
participating hospitals, the Clinical Data Center at Massachusetts General Hospital and the National Institutes of 
Health. 
Risks that are not known: 
We do not know if your blood pressure will be higher or lower or if the function of your lungs and kidneys will be 
better or worse if you choose to be in this study. We also do not know whether your risk of dying will be higher 
or lower if you choose to be in this study.  If you are in the group that receives vitamin C or the group that 
receives acetaminophen, there may be unknown side effects. If you are in the group that receives placebo, you 
will not get the possible benefit of vitamin C or acetaminophen that this study is testing. If we learn something 
new that affects the risks or benefits of being in this study, we will change the study plan as needed. We will also 
make sure you are told about this new information while you are in the study. 
Also, because acetaminophen is being used as one of the treatments in this trial, fever cannot be treated with 
this medication. If you develop a fever, other medicines and cooling methods will be used if your doctors feel 
your fever should be treated, which may be different from standard practice. 
Good effects that might result from this study  
Early evidence suggests that both vitamin C and acetaminophen may improve recovery from sepsis, although we 
are not sure of this. We cannot know whether the vitamin C and acetaminophen will make your recovery better, 
worse, or the same. Your participation in this study helps us learn more about treating patients like you with 
sepsis and may help other patients in the future. The tests we do on your blood will not help you directly.  
During the study: 
 We will talk with your doctors. The research team will tell your doctors about the study, and your doctors 
will decide on all of your additional treatments based on your needs.  
 We will review your medical record. We will look at your medical records or check on you to see how you 
are doing.   We will collect information like your blood pressure, medical history, and test results. We may 
ask questions to you or your doctors if something is not clear. If you go to another health care facility, we 
may contact you or the health care facility to find out how you are doing. 
Date of IRB Approval: 07/14/2021
Date of Expiration: 07/13/2022


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  06/21/2021 
  
 
5 
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 We will collect some of your blood. We will 
collect up to 2 tablespoons of blood at the 
start of the study. We will also collect up to 1 
tablespoon of blood after 2 days, and then up 
to 2 tablespoons of blood after 3 days in the 
study. Some of the blood samples will be 
stored for future studies of serious illness and 
other conditions. Samples will not have your 
name on them, and instead use a coded study number to protect your privacy. We will not include your 
name or other identifying information with the samples ever. Only your hospital study team and the 
research study Coordinating Center at Massachusetts General Hospital will know your coded study number. 
These numbers will be kept private.  
 We will collect some of your urine. We will collect some of your urine (pee) three times during the study: at 
the start of the study, and on days 2 and 3. These samples will be labeled with a coded study number and 
kept private; the same as the blood samples.   
 We will monitor your liver function tests. If you are in the Acetaminophen-ACTIVE or Acetaminophen-
Placebo group, we will measure levels of two liver function tests at the start of the study, and then after 2, 3, 
4, 5, and 7 days in the study. We do not anticipate that this will require an extra blood draw because this 
testing can usually be done in the clinical laboratory at your hospital on blood samples that your regular 
doctors are already sending. Rarely, this may require an extra blood draw. We are doing this because 
acetaminophen can affect the function of your liver. 
 We will check to see how you are doing in one month and in three months. An important part of this study 
is to find out how you are doing after you leave the hospital. To make sure we can reach you, we will record 
your information, such as contact phone numbers and social security number. This information will be kept 
private. 
o If you are still in the hospital, we will check on you in person or by telephone. 
o If you are out of the hospital, we will contact you using the contact information you provide. 
o If we cannot reach you, we will check hospital and national health records using the information you 
provide. 
Other treatments you could get if you decide not to be in this study 
Taking part in this study is voluntary. You will receive the care you need whether or not you join the study or if 
you withdraw at any time. Acetaminophen or vitamin C are not given to all sepsis patients outside of research, 
and if you do not join, your doctor will choose all therapies.  
Reasons why the study doctor may take you out of this study 
The study doctor may take you out of this study if his/her judgment suggests that is better for you. This decision 
may be based on new information about your condition or the study risks and benefits. 
 
 
STUDY BLOOD COLLECTION PLAN: 
Date of IRB Approval: 07/14/2021
Date of Expiration: 07/13/2022


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  06/21/2021 
  
 
6 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
What will happen if you decide to stop being in this study? 
You can stop being in this study at any time. If you decide to stop, tell your study doctor or a member of the 
study team. Deciding to not be part of the study is completely up to you. This will not affect the care you 
receive. 
If you stop being in the study, we will store your blood samples unless you request they be destroyed. We will 
continue to collect information from your medical record unless you ask us not to.  
Clinical Trials Registry  
A description of this clinical trial is available on www.clinicaltrials.gov, which will not include information that 
can identify you. Eventually, the Web site will include a summary of the study results but no personal 
information. You can search this Web site at any time. 
Privacy: 
Any samples and information about you may be made available to others to use for research.  To protect your 
privacy, we will not release your name.  You will not receive any benefit as a result of the tests done on your 
samples.  These tests may help us or other researchers learn more about the causes, risks, treatments, or how to 
prevent this and other health problems. 
Study Results:  
We will share the study results with you by letter within 3 months of the publication of the manuscripts 
reporting the results of the trial.   
 
Date of IRB Approval: 07/14/2021
Date of Expiration: 07/13/2022

Prot_000.pdf: 
 
 
 
Acetaminophen and Ascorbate in 
Sepsis: Targeted Therapy to 
Enhance Recovery (ASTER) 
 
 
 
ASTER: A PETAL Network Multi-Center Phase 2b Randomized Double-Blinded Placebo-
Controlled Trial of Two Different Pharmacologic Therapies (Intravenous Vitamin C or 
Intravenous Acetaminophen)  
VERSION: 4.2  June 8, 2021  
PROTOCOL CO-CHAIRS  
  
Lorraine Ware, MD and Michael Matthay, MD (Co-Chairs Acetaminophen)  
Berry Fowler, MD and D. Clark Files, MD (Co-Chairs Vit C)  
  
PROTOCOL COMMITTEE  
  
 
               
 ADVISORS TO PROTOCOL  
Michael Aboodi, MD  
  
Rebecca Baron, MD   
                                            Lora Reineck, MD  
Samuel Brown, MD    
  
Steven Chang MD, PhD  
Ivor Douglas, MD  
  
                             Roy G Brower, MD  
Abhijit Duggal, MD 
Scott Fields, PharmD  
Adit A. Ginde, MD  
Andrew Goodwin, MD 
Duncan Hite, MD  
Estelle Harris, MD  
Greg Hendey, MD  
Terri Hough, MD   
Akram Khan, MD  
Lori Kozikowski, RN  
Kathleen Liu, MD, PhD  
Chadwick Miller, MD  
Bryce RH Robinson, MD  
Wesley H. Self, MD  
B. Taylor Thompson, MD 
Mark Tidswell, MD  
Donald M. Yealy MD  
STUDY DRUGS  
       Acetaminophen and Vitamin C   
 
 
 
 
 
 
 


  
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Contents 
1. Abbreviations and Definitions .................................................................................................................. 6 
1.1 Abbreviations.......................................................................................................................................... 6 
1.2 Definitions ............................................................................................................................................... 8 
2. Trial Summary ......................................................................................................................................... 10 
2.1 Title ....................................................................................................................................................... 10 
2.2 Objective ............................................................................................................................................... 10 
2.3 Hypotheses ........................................................................................................................................... 10 
2.4 Study Design ......................................................................................................................................... 10 
2.4.1 Treatment Arms .................................................................................................................... 11 
2.4.2 Sample Size/Statistical Considerations: ................................................................................ 12 
2.5 Inclusion Criteria ................................................................................................................................... 12 
2.6 Exclusion Criteria .................................................................................................................................. 12 
2.7 Primary Efficacy Variable ...................................................................................................................... 13 
2.8 Secondary Clinical Efficacy Variables .................................................................................................... 14 
2.9 Safety Variables .................................................................................................................................... 14 
2.10 Biological Endpoints ............................................................................................................................ 15 
3. Trial Description ...................................................................................................................................... 15 
3.1 Background ........................................................................................................................................... 15 
3.1.1 Background-Sepsis and sepsis-related organ dysfunction ................................................... 15 
3.1.2       Cell-free hemoglobin (CFH) and organ dysfunction in sepsis ......................................... 16 
3.2      Vitamin C—Preclinical and Clinical Data .......................................................................................... 16 
3.3     Acetaminophen—Preclinical and Clinical Data ................................................................................. 19 
3.4 Summary ............................................................................................................................................... 21 
4. Trial Design ............................................................................................................................................. 22 
4.1 Study Description ................................................................................................................................. 22 
4.2 Randomization ...................................................................................................................................... 23 
4.3 Primary Efficacy Variable ...................................................................................................................... 24 
4.4 Secondary Clinical Efficacy Variables .................................................................................................... 24 


  
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4.5 Safety Variables .................................................................................................................................... 25 
4.6 Biological Endpoints .............................................................................................................................. 25 
5. Treatments Arms and Study Drug Administration ................................................................................. 26 
5.1 Acetaminophen-Active Arm ................................................................................................................. 26 
5.2 Acetaminophen-Placebo Arm ............................................................................................................... 27 
5.3 Vitamin C Arm-Active ............................................................................................................................ 27 
5.4 Vitamin C-Placebo Arm ......................................................................................................................... 28 
5.5 Drug Interruptions ................................................................................................................................ 28 
6. Study Population and Enrollment ........................................................................................................... 28 
6.1 Participant Enrollment .......................................................................................................................... 28 
6.2 Inclusion Criteria ................................................................................................................................... 29 
6.3 Exclusion Criteria .................................................................................................................................. 29 
6.4     Reasons for each of the exclusion criteria ........................................................................................ 30 
6.5 Co-enrollment ....................................................................................................................................... 31 
6.6 Informed Consent ................................................................................................................................. 31 
6.7 Excluded Medications ........................................................................................................................... 32 
6.8 On-study Fever Management Recommendations................................................................................ 32 
6.9 Concomitant Medications .................................................................................................................... 32 
6.10 Blood Glucose Strategy for the Vitamin C-Active/Vitamin C-Placebo Arm ........................................ 33 
6.11 Common Strategies for All Treatment Arms ...................................................................................... 33 
6.11.1 Ventilator Procedures ......................................................................................................... 33 
6.11.2   On Study Fluid Management ............................................................................................ 33 
6.12 Criteria for Discontinuation or Withdrawal ........................................................................................ 34 
7.0 Study Plan ............................................................................................................................................. 34 
7.1 Baseline Assessments ........................................................................................................................... 34 
7.2. Assessments During Protocol Phase (through study day 7) ................................................................ 35 
7.3. Other Assessments and Data Collection .............................................................................................. 35 
7.4. Assessments after Hospitalization ....................................................................................................... 36 
7.5. 90-day All-Cause Mortality .................................................................................................................. 36 
7.6. Specimen Collection ............................................................................................................................ 36 
8. Statistical Methods ................................................................................................................................. 37 
8.1 Primary Endpoint .................................................................................................................................. 37 


  
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8.2   Sample Size ......................................................................................................................................... 37 
8.3 Secondary endpoints ............................................................................................................................ 38 
8.4 Safety Endpoints ................................................................................................................................... 40 
8.5 Predictive Biomarker  Analysis ............................................................................................................. 40 
8.6 Statistical Analysis Plan ......................................................................................................................... 40 
8.6.1 Sub-Group by Treatment Interaction ................................................................................... 40 
8.7 Interim Analysis .................................................................................................................................... 40 
9.0 Risk Assessment .................................................................................................................................... 41 
9.1 Potential Risks to Subjects .................................................................................................................... 41 
9.2  Risk of Death ........................................................................................................................................ 42 
9.3  Minimization of Risk ............................................................................................................................ 43 
9.4 Potential Benefits ................................................................................................................................. 43 
9.5 Risks in Relation to Anticipated Benefits .............................................................................................. 43 
10. Human Subjects .................................................................................................................................... 43 
10.1 Selection of Subjects ........................................................................................................................... 43 
10.2   Justification for Including Vulnerable Subjects ................................................................................. 43 
10.3 Informed Consent ............................................................................................................................... 44 
10.3.1 Process of Obtaining Informed Consent ............................................................................. 44 
10.4   Continuing Consent ........................................................................................................................... 46 
10.5 Withdrawal of Consent ....................................................................................................................... 46 
10.6    Identification of Legally Authorized Representatives ...................................................................... 46 
10.7    Justification of Surrogate Consent ................................................................................................... 47 
10.8     Additional Safeguards for Vulnerable Subjects ............................................................................... 47 
10.9     Confidentiality ................................................................................................................................. 48 
11. Adverse Events ..................................................................................................................................... 48 
11.1 Safety Monitoring ............................................................................................................................... 48 
11.2 Serious Adverse Events ....................................................................................................................... 49 
Appendices ................................................................................................................................................. 50 
A. Schedule of Events ................................................................................................................................. 50 
B. Sequential (Sepsis-related) Organ Failure Assessment (SOFA) Score .................................................... 51 
C. Ventilator Management for Patients with ARDS .................................................................................... 52 
D. On-Study Fluid Management for Patients with ARDS ............................................................................ 54 


  
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E. Adverse Event Reporting and Unanticipated Events .............................................................................. 55 
E.1. Unanticipated Problems (UP) ................................................................................................. 55 
E.2. Determining Relationship of Adverse Events to Procedures .................................................. 55 
E.3. Clinical Outcomes that may be Exempt from Adverse Event Reporting ................................ 56 
F. Alcohol Use Screen Audit C ..................................................................................................................... 57 
G. Fever Management Guidelines .............................................................................................................. 58 
References .................................................................................................................................................. 59 
 
  
  


  
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1. Abbreviations and Definitions   
1.1 Abbreviations   
ABG = Arterial Blood Gas  
APAP = Acetaminophen  
AKI = Acute Kidney Injury  
ALI = Acute Lung Injury  
ALT = Alanine Aminotransferase  
ARDS = Acute Respiratory Distress Syndrome  
AST = Aspartate Aminotransferase  
BIPAP = Bi-level Positive Airway Pressure  
BMI = Body mass index  
BP = Blood Pressure  
CCC = Clinical Coordinating Center  
CFH = Cell-free hemoglobin  
CPAP = Continuous Positive Airway Pressure  
CRF = Case Report Form  
Day 0 =  Day of Randomization  
DHA = Dehydroascorbic Acid  
DSMB = Data and Safety Monitoring Board  
ELISA = Enzyme-linked Immunosorbent Assay  
  
  
 
 
 
 
 
 
 
ED = Emergency Department  
F2-IsoP = F2-Isoprostanes  
FDA = Food and Drug Administration  
FiO2 = Fraction of Inspired Oxygen  
GCS = Glasgow Coma Scale  
HFNO = High Flow Nasal Oxygen  
HUVEC = Human Umbilical Vein  
Endothelial Cells  
ICF = Informed Consent Form  
ICU = Intensive Care Unit  
IMV = Intermittent Mechanical Ventilation  
IL-6 = Interleukin-6  
IL-8 = Interleukin-8  
IRB = Institutional Review Board  
IV = Intravenous  
LAR = Legally Authorized Representative  
LPS = Lipopolysaccharide  
MAP = Mean arterial blood pressure  
MBW = Measured Body Weight  
mL = Milliliter  
mmHg = Millimeter of Mercury  
 
MAKE28 = Major Adverse Kidney Events at 
28 days  


  
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NHLBI = National Heart Lung and Blood  
Institute  
NIV = Non-invasive ventilation   
NSAID = Nonsteroidal anti-inflammatory drug  
PaO2 = Partial pressure of arterial oxygen  
PBW = Predicted Body Weight  
PEEP = Positive End-Expiratory Pressure   
Pplat = Plateau pressure  
PSV = Pressure Support Ventilation  
POC = Point of Care  
RALE = Radiographic Assessment of Lung Edema  
RCT = Randomized Controlled Trial  
RRT = Renal Replacement Therapy  
SAE = Serious Adverse Event  
SBP = Systolic Blood Pressure  
SBT = Spontaneous Breathing Trial  
SOFA = Sequential (Sepsis-related) Organ Failure 
Assessment   
SpO2 = Oxygen Saturation by Pulse  
Oximetry  
SUSAR = Suspected and Unexpected Serious 
Adverse Reaction  
UAB = Unassisted Breathing  
VFD = Ventilator-free Days  
Vit C = Ascorbic acid (Vitamin C)  
  
  
  
  
  


  
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1.2 Definitions  
Adverse Event: Any untoward medical occurrence associated with the use of a drug or a study 
procedure, whether or not considered drug related.  
Adverse Reaction: Any adverse event caused by a study intervention. An adverse reaction is a 
subset of all suspected adverse reactions where there is a reason to conclude that the study 
intervention caused the event.   
Assisted ventilation: Any level of invasive or noninvasive ventilator support including 
continuous positive airway pressure of >5 cmH20, bilevel positive pressure ventilation, or 
invasive mechanical ventilation.  Noninvasive ventilation solely for sleep disordered breathing 
and nasal high flow oxygen therapy are not considered “assisted ventilation” for this study.  
Baseline: Prior to randomization  
Chronic Dialysis: Receipt of hemodialysis or peritoneal dialysis for renal failure prior to 
hospitalization.  This does not include new renal replacement therapy or hemofiltration initiated 
within 36 hours of randomization. 
Date of first Unassisted Breathing (UAB): Defined as the first day that the subject is on UAB 
from midnight to midnight. Example: if subject meets UAB at 1900 on 6/1/14, then the date of 
first UAB would be 6/2/14, as long as subject does not return to AB on 6/2/14.  
Day zero: Defined as the day of randomization (until 23:59 of the study day).  
Days Alive and Free of Organ Support: Days alive and free of assisted ventilation, 
vasopressors and new renal replacement therapy over 28 days  
Extubation: Removal of an orotracheal or nasotracheal tube, or unassisted breathing with a 
tracheostomy  
Fluid Bolus: 500 ml or more of isotonic crystalloid  
High Flow Nasal Oxygen: Oxygen delivered by nasal cannula at a rate of at least 30 liters/min, 
using a system capable of delivering up to 100% humidified and heated oxygen at flow rates up 
to 60 liters per minute.  
Home: Level of residence or health care facility where the patient was residing prior to hospital 
admission  
Invasive Mechanical Ventilation: Mechanical ventilation delivered through an endotracheal or 
tracheostomy tube.  
Legally Authorized Representative: Any individual, judicial, or other body authorized under 
applicable law to consent on behalf of a prospective patient to the patient’s participation in the 
clinical study  
Serious Adverse Event: Adverse events that are serious and have a reasonable possibility that 
the event was due to a study drug or procedure.  


  
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Pg.  9     
Study hospital: Defined as the hospital where the patient was randomized and enrolled.  
Study withdrawal: Defined as permanent withdrawal from study the before completion of study 
activities. This does not include those subjects who have completed the protocol procedures. If 
a patient or surrogate requests withdrawal from the study the investigator should seek explicit 
permission to continue data collection.  
Suspected Adverse Reaction: Any adverse event for which there is a reasonable possibility 
that the study procedures caused the adverse event. Reasonable possibility means there is 
evidence to suggest a causal relationship between the study procedures and the adverse event. 
A suspected adverse reaction implies a lesser degree of certainty about causality than adverse 
reaction.   
Suspected Unexpected Serious Adverse Reaction (SUSAR): An adverse reaction that is 
both unexpected (not consistent with risks outlined in the protocol or investigator brochure), 
serious, and meets the definition of a suspected adverse reaction.  
UAB (Unassisted Breathing):  Spontaneously breathing with face mask, nasal prong oxygen,  
or room air, T-tube breathing, tracheostomy collar (mask) breathing, or CPAP ≤ 5 without PSV 
or IMV assistance, nasal high flow oxygen, or the use of noninvasive ventilation solely for sleep-
disordered breathing.  Assisted breathing is any level of ventilatory support at pressures higher 
than the unassisted breathing thresholds.  
 
 


  
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2. Trial Summary  
2.1 Title  
Acetaminophen and Ascorbate in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
ASTER is a platform on which we will conduct two separate trials.  
2.2 Objective  
Objective: To carry out two multi-center phase 2b randomized double-blinded placebo-
controlled trials of two different pharmacologic therapies. One trial will assess the efficacy of 
Acetaminophen (1 gram intravenously every 6 hours) in comparison to placebo for 120 hours in 
patients with sepsis who have evidence of either hemodynamic or respiratory organ failure. A 
second trial will assess the efficacy of Vitamin C (50 mg/kg every 6 hours) compared to placebo 
infused intravenously for 120 hours in patients with sepsis who have evidence of either 
hemodynamic or respiratory organ failure. The two placebo infusions (5% dextrose in water, 
D5W) will match the volume and storage temperatures of the acetaminophen (room 
temperature, dose adjusted for weight below 50 kg) and Vitamin C (room temperature, volume 
adjusted for weight above and below 80 kg with maximum dose of 9 grams every 6 hours).  For 
efficiency, both the Acetaminophen and the Vitamin C placebo groups will be pooled for the 
efficacy comparisons. All arms will have the same inclusion and exclusion criteria.   
2.3 Hypotheses  
Hypothesis 1A: Acetaminophen (APAP) or Vitamin C infusion will increase the days alive and 
free of organ support to day 28.  
Hypothesis 1B: APAP or Vitamin C will have a favorable effect on other secondary outcomes 
including pulmonary and non-pulmonary organ dysfunction and biomarkers of inflammation and 
endothelial injury  
2.4 Study Design  
Prospective multi-center phase 2b randomized placebo-controlled double-blinded interventional 
trials of two different pharmacologic therapies (intravenous Vitamin C or intravenous 
Acetaminophen) for patients with sepsis-induced hypotension or respiratory failure.   
1. Enrollment Period: Approximately 15 months  
2. Patient Population: Critically ill adults admitted or planned admission to the ICU with 
sepsis and either hemodynamic or respiratory organ failure.  A maximum of 900 patients 
will be enrolled  
3. Locations: Emergency Department and ICUs at hospitals participating in the NHLBI 
PETAL Network  


  
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2.4.1 Treatment Arms 
Patients with sepsis and either hemodynamic or respiratory organ failure on enrollment will be 
randomized to one of four treatment arms in a 2:1:2:1 fashion.  The four arms will be 
Acetaminophen (APAP)-Active, Acetaminophen (APAP)-Placebo, Vitamin C-Active, and Vitamin 
C-Placebo, randomized in a 2:1:2:1 randomized ratio in order to maximize efficiency and 
maintain blinding for the different study drug storage conditions.     
 
At the time of randomization, the research team will be notified that the patient is randomized into 
the APAP-Active/APAP-Placebo trial or the Vitamin C-Active/Vitamin C-Placebo trial.  The 
patients, nurses, research staff, and physicians will be blinded to the treatment assignment within 
each trial.  The time of randomization will represent time zero.  Study drug will be started within 4 
hours of randomization.  
 
 
 
 
 
 
 
 
 
 
Treatment will 
continue for 120 
hours (20 doses), or 
discharge from the 
intensive care unit, 
study withdrawal, or 
death, whichever 
comes first. 
First study drug dose  
(APAP, Vit C, or placebo) 
will be considered “Dose 1” 
and will be administered 
within 4 hours of 
randomization.   
Study drug will be infused 
over 30 minutes.  
 
Subsequent doses will be infused 
every six hours (+/- 1 hour) 
through 120 hours or 20 doses 
(whichever comes first).  If a dose 
cannot be administered within 3 
hours of the scheduled time, the 
dose should be skipped.   
Any dose administered outside of 
the +/- 1 hour window or any dose 
that is skipped will trigger a protocol 
deviation 
Duration 
Subsequent Dosing 
Timing of Dosing 
3. Vitamin C-Active given intravenously at the dose of 50 mg/kg every six hours (to a 
maximum dose of 9 grams every six hours) for 5 days (20 doses) OR 
1. APAP-Active given intravenously at the dose of 1 gram (or 15 mg/kg if patient weighs 
< 50 kg) every six hours for 5 days (20 doses) OR 
2. APAP-Placebo (identical appearing 5% dextrose solution) infused every six hours for 5  
days (20 doses) OR 
4. Vitamin C-Placebo  (identical appearing 5% dextrose solution) infused every six hours 
for 5 days (20 doses)   
NOTE: The volume of APAP-Placebo or Vitamin C-Placebo infusions will be lower in 
patients <50kg (APAP) or <80 kg (Vitamin C) and will be matched in their respective 
placebos (see pharmacy manual). 


  
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Completion of study drug administration: Study drug administration will occur for a total of 
120 hours, unless one of the following occurs:  
1. Discharged from the study hospital   
2. Discharge from the ICU  
3. Withdrawal from the study   
4. Death  
5. New AST or ALT elevation ≥ 10 times the upper limit of normal (for the APAP-
Active/APAP-Placebo group only)  
2.4.2 Sample Size/Statistical Considerations:  
With randomization of 900 total patients (300 Acetaminophen-Active, 300 Vitamin C-Active, 150  
Acetaminophen-Placebo, 150 Vitamin C-Placebo ) with pooled placebo groups, we will have 
85% power to detect a difference between groups of 2.5 days in the primary composite outcome 
of days alive and free of organ support (dialysis, assisted ventilation, and vasopressors) to day 
28. Patients will be analyzed on an intention-to-treat basis based on randomization assignment.  
 
2.5 Inclusion Criteria  
1. Age ≥ 18 years  
2. Sepsis defined as:  
a. Clinical evidence of a known or suspected infection and orders written to administer 
antibiotics  
AND  
b. Hypotension as defined by the need for any vasopressor (and at least 1 liter of fluid 
already administered intravenously for resuscitation) OR respiratory failure defined 
by mechanical ventilation,  BIPAP or CPAP at any level, or greater than or equal to 6 
liters/minute of supplemental oxygen (criterion b must be present at the time of 
randomization)  
3. Admitted to a study site ICU (or intent for the patient to be admitted to a study site ICU) 
within 36 hours of presentation to the ED or admitted to the study site ICU within 36 hours 
of presentation to any acute care hospital  
2.6 Exclusion Criteria  
1. No consent/inability to obtain consent from the participant or a legally authorized 
representative  
2. Patient unable to be randomized within 36 hours of presentation to the ED or within 36 hours 
of presentation to any acute care hospital  
3. Diagnosis of cirrhosis by medical chart review  
4. Liver transplant recipient   
5. AST or ALT greater than five times the upper limit of normal   


  
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6. Diagnosis of ongoing chronic alcohol use disorder/abuse by chart review; if medical record 
unclear, use Appendix F  
7. Clinical diagnosis of diabetic ketoacidosis or other condition such as profound hypoglycemia 
that requires hourly blood glucose monitoring  
8. Hypersensitivity to Acetaminophen or Vitamin C  
9. Patient, surrogate or physician not committed to full support (Exception: a patient will not be 
excluded if he/she would receive all supportive care except for attempts at resuscitation from 
cardiac arrest)  
10. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used 
only for sleep-disordered breathing  
11. Chronic dialysis 
12. Current active kidney stone   
13. Multiple (>1) episodes of prior kidney stones, known history of oxalate kidney stones, or 
history of oxalate nephropathy  
14. Kidney transplant recipient   
15. Use of home oxygen for chronic cardiopulmonary disease  
16. Moribund patient not expected to survive 24 hours   
17. Underlying malignancy or other condition with estimated life expectancy of less than 1 month  
18. Pregnant woman, woman of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or woman who is breast feeding  
19. Prisoner  
20. Treating team unwilling to enroll because of intended use of Acetaminophen or Vitamin C 
21. Treating team unwilling to use plasma (as opposed to point of care testing) for glucose 
monitoring 
2.7 Primary Efficacy Variable  
Primary efficacy variables will be evaluated comparing Acetaminophen vs. pooled placebo and 
Vitamin C vs. pooled placebo in separate analyses. The primary efficacy variable is days alive 
and free of organ support (dialysis, assisted ventilation, and vasopressors) to day 28.  
Participants will need to be free of all three components (assisted ventilation, vasopressors, new 
renal replacement therapy) to qualify for a day alive and free from organ failures. A patient could 
be free of all organ failures from day 10 through day 22 and then die on day 22 and would 
receive 12 days of credit for this primary outcome.  The components of this outcome (days free 
of dialysis, assisted ventilation, and vasopressors in the overall cohort and in survivors, and 28-
day hospital mortality) will also be reported as secondary outcomes (see below in 2.8).  


  
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2.8 Secondary Clinical Efficacy Variables  
Secondary efficacy variables will be evaluated comparing Acetaminophen vs. pooled placebo 
and Vitamin C vs. pooled placebo in separate analyses.  
1. 28-day ventilator-free days   
2. 28-day vasopressor-free days   
3. 28-day new renal replacement-free days  
4. 28-day hospital mortality   
5. 28-day ICU free-days  
6. 28-day hospital-free days to discharge home  
7. 28-day duration of ICU stay in survivors and non-survivors   
8. Initiation of assisted ventilation to day 28  
9. Initiation of renal replacement therapy to day 28  
10. Change in Sequential Organ Failure Assessment (SOFA) scores between enrollment 
and study day 7  
11. 90-day hospital mortality (includes death at any healthcare facility that is not the 
equivalent of home for a patient)  
12. Development of ARDS within 7 days of randomization   
13. Change in serum creatinine from enrollment to discharge, death, initiation of dialysis or 
28 days, whichever occurs first   
14. Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline, need for new renal replacement therapy, or death  
15. Change in the Radiographic Assessment of Lung Edema (RALE) score from enrollment 
to  study day 3 in patients who are receiving assisted ventilation or high flow nasal 
oxygen at baseline.  
16. 90-day all-cause mortality   
17. Renal calculi diagnosed between randomization and study day 90 in the Vitamin C-
Active/Vitamin C-Placebo arm  
2.9 Safety Variables  
The safety monitoring and analyses will be performed separately on the APAP-Active/APAP-
Placebo arm and the Vitamin C-Active/Vitamin C-Placebo arms (2 active to 1 placebo 
randomization ratio). The safety variables will include the following:  
1. Hypersensitivity or rash  
2. AST and ALT measured on study days 0, 2-5, and 7 (day 7 measurement can be 1 day) 
in the APAP-Active/APAP-Placebo arm only.  


  
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3. The plasma levels of Acetaminophen will be obtained as a trough level just prior to 
administration of a study dose on study day 2 (subject must have received at least 5 doses 
of study drug before the day 2 samples are obtained) in the APAP-Active/APAP-Placebo 
arm only. After the first 100 patients have been enrolled who have been randomized to the 
Acetaminophen arm, the trough levels will be reported to the DSMB to determine if they 
exceed the level of 20 micrograms/mL, the potential level for hepatotoxicity.  
4. Administration of fluid bolus, new use of vasopressor, or increased dose of vasopressor 
within 120 minutes of study drug infusion in the APAP-Active/APAP-Placebo arm only.  
5. Incidence of reported adverse events  
2.10 Biological Endpoints  
Change in plasma IL-6, angiopoietin-2, cell-free DNA, syndecan-1, and cell-free hemoglobin 
between enrollment (baseline) and Days 2 and 3.  
3. Trial Description  
3.1 Background  
3.1.1 Background-Sepsis and sepsis-related organ dysfunction  
Sepsis is a common condition with over 750,000 cases per year in the U.S. and an associated 
mortality of at least 18-38% [1, 2].  The primary cause of death in sepsis is organ dysfunction, 
including acute respiratory distress syndrome (ARDS), acute kidney injury (AKI) and 
cardiovascular dysfunction.  ARDS is a common complication of sepsis; in a large multicenter 
international study of ARDS, sepsis (pulmonary or extrapulmonary) was the most common 
cause of ARDS [3].  Mortality from sepsis-induced ARDS is higher than mortality from ARDS 
due to other causes [4]. AKI is also common in sepsis, occurring in greater than 50% of patients 
[5-7]; the range of morbidity associated with AKI is broad, including prolonged ICU length of 
stay, need for renal replacement therapy (RRT), and death. Furthermore, ARDS and AKI 
commonly occur together, suggesting overlapping pathophysiologic mechanisms.  The 
development of AKI in patients with ARDS increases mortality [8]. However, despite the 
magnitude of this clinical problem, there are still no specific therapies for sepsis other than 
antimicrobials, and no specific therapies that reduce the severity of sepsis-associated ARDS, 
AKI or cardiovascular dysfunction [9].  
 


  
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 3.1.2       Cell-free hemoglobin (CFH) and organ dysfunction in sepsis  
Sepsis causes alterations in the red blood cell membrane that lead to release of CFH into the 
circulation due to membrane damage [10-12].  In an animal model of cecal-ligation and puncture 
induced sepsis, CFH can be measured in the plasma and levels of CFH are associated with 
mortality and worse renal function, an effect that is potentiated by deficiency of haptoglobin, 
hemopexin, or heme oxygenase-1[13]. In critically ill adults with sepsis, CFH is present in 80%-
90% of patients, is associated with oxidative injury as measured by plasma F2-isoprostanes (F2-
IsoP), and is independently associated with an 
increased risk of in-hospital mortality [14, 15] 
(Figure 1).    
Mechanisms of toxicity of CFH in sepsis include 
the ability of CFH to scavenge nitric oxide, 
leading to vasoconstriction in various vascular 
beds [16-19], proinflammatory effects of CFH 
and its heme subunit [20-22] and the ability of 
CFH to undergo redox cycling, leading to 
oxidation of lipid membranes and release of F2-
IsoP [23, 24]; plasma levels of F2-IsoP, are 
increased in patients with sepsis and are 
associated with renal, hepatic, and coagulation 
failure, and mortality [25]. This trial will test two 
agents, Vitamin C and Acetaminophen, that can 
specifically target the injurious effects of CFH, 
present in the vast majority of patients with 
sepsis, to reduce organ dysfunction and affect 
clinical outcomes. To determine whether pre-
enrollment point-of-care measurement of plasma CFH can be used for predictive enrichment in 
a future phase 3 trial of either of these agents, samples will be collected at enrollment in the 
current trial for post hoc measurement of plasma CFH to determine what level of pre-enrollment 
plasma CFH is optimal for predictive enrichment.    
3.2      Vitamin C—Preclinical and Clinical Data  
To study the effect of Vitamin C infusion on systemic inflammatory responses and acute organ 
injury associated with bacterial sepsis, a durable model of septic shock and acute lung injury in 
C57BL6 mice was created to determine the extent to which Vitamin C could be employed as an 
interventional (rather than preventive) therapy. Sepsis was induced by intraperitoneal (IP) 
administration of E coli lipopolysaccharide (LPS, 0111:B4) at a concentration of 10 mcg/gram of 
body weight or feces-induced peritonitis.  In both models, Vitamin C infusion had beneficial 
effects on multiple endpoints including attenuation of the proinflammatory response, 
enhancement of epithelial barrier function, increasing alveolar fluid clearance, and prevention of 
sepsis-associated coagulation abnormalities [26, 27].  In the E Coli LPS model, Vitamin C or its 
DHA derivative had a favorable impact on survival (Figure 2). These studies show that Vitamin 
C has a favorable impact on both acute lung injury and survival in mouse models of sepsis. 
Figure 1.  Plasma  cell - free hemoglobin levels are  
significantly associated with in-hospital mortality in 391  
critically ill patients with sepsis (14)  


  
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Interestingly, one mechanism may be through the 
capacity of Vitamin C to block CFH-mediated endothelial 
injury.  In cultured human umbilical vein endothelial cells 
(HUVECs), CFH increased endothelial monolayer 
permeability in a dose- and time-dependent manner and 
caused a dramatic reduction in intracellular ascorbate. 
Pretreatment of HUVECs with Vitamin C markedly 
attenuated CFH-induced permeability [28].   
Vitamin C has been intravenously infused in humans in 
high dosages in several studies. Nathens et al 
administered 1.5 g of Vitamin C IV every 8 hours (total 
dose of 4.5 grams per day) in surgical trauma ICU 
patients daily for 28 days with no adverse events [29]. 
Muhlhofer et al infused high dose ascorbic acid (7.5 g) intravenously into human volunteers 
daily for 7 days with no adverse events [30]. Finally, Tanaka et al infused high-dose ascorbic 
acid (66 mg/kg/hour, average 110 grams for 70 kg human) for 24 hours into patients with over 
50% total body surface area burns [31]. No abnormalities in hematologic, hepatic, or renal 
function were seen at day 7 following ascorbate administration.  In 2010, the Fowler group 
completed a phase I, randomized, double blind, placebo-controlled trial testing the safety of 
parenteral Vitamin C in patients with severe sepsis [32]. Patients were randomized to placebo 
(5% dextrose/water, D5W), lower dose Vitamin C (50 mg/kg/24hr), or higher dose Vitamin C (50 
mg/kg every 6 hours). The calculated 24-hour Vitamin C dose was divided into four equal doses 
and administered IV (in 50 ml 5% dextrose in water) over 30 minutes every 6 hours for 96 
hours.  During the infusion period, no patients were withdrawn due to adverse effects 
(hypotension, tachycardia, nausea/vomiting, or hypernatremia). Infusion was halted in one 
patient (Hi Vitamin C) for a ventricular arrhythmia later determined to be artifact. Figure 3 shows 
that patients treated with high dose of Vitamin C exhibited significantly lower SOFA scores when 
compared to placebo over the 96 hour period. There were also trends for fewer ICU days and 
more VFD in the Low Vitamin C patients, but the numbers were small and statistically 
insignificant. There were no significant differences in mortality.  In this trial, high dose 
intravenous Vitamin C in patients with sepsis-associated 
ARDS significantly reduced cell-free DNA and 
Syndecan-1 plasma levels at 48 hours.  
Given the results of the phase I safety trial, a multi-
center phase 2 randomized double-blind placebo 
controlled NHLBI funded clinical trial (CITRIS-ALI) was 
conducted from September 2014 to January 2018 to 
determine if IV infusion of Vitamin C at 50 mg/kg every 6 
hours for 96 hours (same as higher dose in the phase 1 
trial, i.e. a total of 200 mg/kg/24 hours) would affect the 
primary outcomes of SOFA score at 96 hours and 
plasma biomarkers of systemic inflammation (C-reactive 
protein) and vascular injury (thrombomodulin). The study 
enrolled 167 patients with sepsis-associated ARDS; 83 
patients were randomized to placebo, and 84 to IV Vitamin C.   
Figure 3.    SOFA scores in sepsis  
patients were significantly lower over  
time in the high dose Vitamin C  
treatment group compared to placebo   
Time (h) 
Figure 2.    Vitamin C (AA) or its DHA  
derivative significantly improved survival in  
a mouse model of lethal peritonitis induced  
by intraperitoneal injection of LPS   


  
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There was no difference in adverse events or serious adverse events between groups.  Vitamin 
C significantly lowered organ failure scores (SOFA) at 96 hours but not plasma C-reactive 
protein or thrombomodulin compared to placebo in the first 96 hours of ICU care. Vitamin C was 
also associated with beneficial effects on several secondary outcomes.  Vitamin C produced a 
significant increase in ICU-free days and a trend towards more ventilator-free days (Figure 4).   
By Kaplan Meier analysis, IV Vitamin C infused for a total of 96 hours significantly improved 
survival at both day 28 and day 60 (Figure 4).  Vitamin C also significantly reduced plasma 
syndecan-1 and cell-free DNA levels [33].  The dose of 50 mg/kg every 6 hours of Vitamin C 
has been selected for this phase 2b ASTER trial because it was not associated with safety 
issues in the CITRIS-ALI or Fowler’s phase I trials and it was associated with efficacy compared 
to placebo in several secondary endpoints. 
 
Other trials of high-dose Vitamin C are being conducted by the international community. The 
Lessening Organ Dysfunction with Vitamin C (LOVIT, NCT03680274) Trial is a randomized placebo 
controlled trial conducted by the Canadian Critical Care Trials Group [34]. LOVIT enrolls patients with 
septic shock to receive either Vitamin C (50 mg/kg) IV Q 6 hours for 4 days vs placebo. This trial 
began enrollment prior to the COVID-19 pandemic. However, the trial was opened to COVID-19 
positive patients following the onset of the pandemic.  As of February 2021, this trial had enrolled 729 
of 800 planned participants. Two trials of high dose Vitamin C are ongoing that focus exclusively on 
COVID-19 positive acute respiratory failure. The LOVIT COVID trial (NCT04401150) and REMAP-
CAP Vitamin C domain are both evaluating high dose Vitamin C in COVID-19 acute respiratory failure.  
 
Collectively, these data show strong preclinical and early human data to test high dose Vitamin C in 
patients with sepsis and lung injury.  The ongoing trials in both COVID-19 negative and positive 
patients highlight the broad interest in the critical care community for evaluating this therapy.  
Figure 17 
Figure 18 
Figure 19 
A   
  
  
          
B   
  
  
  
  
C   
Figure 4 .  Results of the CITRIS- ALI trial.  Vitamin C had favorable effects on  
ventilator-free days (Panel A), ICU-free days (Panel B) and survival (Panel C) in  
patients with ARDS.  


  
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3.3     Acetaminophen—Preclinical and Clinical Data  
The systemic inflammatory response to infection in sepsis creates an oxidative environment that 
has the potential of oxidizing the iron in CFH from the Ferrous (Fe 2+) to the ferryl (Fe 4+) state, 
creating an iron radical capable of organ injury 
[14, 24, 35]. Acetaminophen, at normal 
therapeutic concentrations, is a potent and 
specific inhibitor of hemoprotein-mediated lipid 
peroxidation owing to its ability to reduce the 
ferryl-protoporphyrin radical generated with 
release of hemoproteins into the circulation 
(Figure 5) [35, 36].  The hemoprotein reductant 
activity of Acetaminophen is independent of its 
other pharmacologic mechanisms, including 
prostaglandin H2 synthetase and 
cyclooxygenase inhibition [36, 37]. In an 
experimental study in rats, Acetaminophen was  
highly effective in preventing oxidative damage 
and renal failure in the setting of myoglobinemia due to rhabdomyolysis  [35].  In an 
observational study of patients with sepsis, receipt of Acetaminophen was associated with 
decreased F2-IsoP levels and independently associated with decreased in-hospital mortality 
[14].  
In the isolated perfused human lung, addition of CFH to the perfusate causes acute lung injury 
manifested by an increase in lung vascular permeability and pulmonary edema formation as 
measured by lung weight gain and increased alveolar capillary barrier permeability to protein as 
measured by accumulation of Evans Blue labeled albumin in the airspaces of the lung.  In this 
human lung preparation, addition of Acetaminophen to the perfusate blocked the effects of CFH 
on vascular permeability and pulmonary edema formation (Figure 6).  
 
Figure  6 .  Addition of  Acetaminophen   ( APAP) to the perfusate blocks the effects of CFH on pulmonary edema formation  
( panel A) and alveolar capillary barrier permeability (panel B) in paired isolated perfused human donor lungs.  
Figure 5 . Release of cell free hemoglobin into an oxidizing  
environment as in sepsis, leads to formation of the highly  
reactive Fe4+ ferryl radical 


  
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The Ware group conducted a phase 2a, randomized, double blind, placebo controlled pilot 
clinical trial of enteral Acetaminophen in 40 critically ill adults with severe sepsis [38] entitled the 
ACROSS I Trial (Acetaminophen for the Reduction of Oxidative injury in Severe Sepsis).  
Patients were enrolled within 24 hours of admission to the ICU.  After enrollment, blood was 
drawn for measurement of plasma CFH to identify patients who may benefit from  
Acetaminophen therapy.  Patients with undetectable CFH (<10 mg/dL) were not randomized,  
while patients with any amount of CFH (≥ 10 mg/dL) were randomized to enteral  
Acetaminophen at the dose of 1 gram enterally every 6 hours or placebo for 3 days.  Notably, 
91% of patients who met inclusion criteria for this trial had detectable CFH and thus were 
eligible for randomization.  The primary endpoint was plasma F2-IsoP on study day 3, with 
secondary endpoints of F2-IsoP on study day 2 and serum creatinine on study day 3.  
On study day 3, F2-Isoprostanes were not significantly lower in the Acetaminophen group (30 
pg/mL, IQR 24-41) compared with placebo (36 pg/mL, IQR 25-80, p = 0.35) although power was 
reduced at that timepoint due to patient dropout.  However, F2-Isoprostanes were significantly 
reduced on study day 2 in the Acetaminophen group (24 pg/mL, IQR 19 – 36) compared with 
placebo (36 pg/mL, IQR 23-55, p = 0.047).  Creatinine on study day 3 was significantly lower in 
the Acetaminophen group (1.0 mg/dL, IQR 0.6–1.4) compared with placebo (1.3 mg/dL, IQR 
0.83 – 2.0, p = 0.039).  This difference persisted beyond study completion (Figure 7A). There 
was no statistically significant difference in hospital mortality (Acetaminophen 5.6% vs. placebo 
18.2%, p = 0.355) although there was a trend for benefit (Figure 7B).  There were no significant 
adverse events (AST or ALT >400, Acetaminophen 9.5% vs. placebo 4.3%, p = 0.599).   
  
Although the reduction in biomarkers of oxidative injury in the study was not consistent across 
all analyses, the biologic plausibility of benefit with Acetaminophen in patients with sepsis and 
detectable CFH, ease and safety of administration, consistent although non-significant 
improvement across all clinical outcomes, and the finding of improved renal function in sepsis 
warrants a larger Phase 2b trial in sepsis patients.  A recent clinical trial of oral Acetaminophen 
in severe falciparum malaria also showed a reduction in creatinine in the treated arm with no 
adverse safety signal, further supporting the need for larger clinical trials [39].    A phase 3 trial 
of acetaminophen for treatment of severe falciparum malaria in the Democratic Republic of 
Congo (NCT04251351) is fully funded with plans to start enrollment in 2021 as soon as COVID-
19 pandemic-related restrictions are lifted. 
  
  
Panel A: 
  
Figure 7, Panel A: Creatinine measured at enrollment, each study day, and after study completion in patients never requiring 
renal replacement therapy. Panel B: Hospital survival by treatment group. 
Panel B: 


  
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Another trial, the ANZICS HEAT trial [40], has some superficial similarities to the 
Acetaminophen arm of the ASTER trial but did not target sepsis patients with organ dysfunction 
and used relatively low cumulative doses of Acetaminophen. In the HEAT trial, critically ill 
patients (n = 690) with fever were randomized to receive IV Acetaminophen or placebo 1g every 
6h for up to 28 days, but only while febrile, leading to a median total dose of only 8g. This dose 
is significantly lower than our proposed total dose of 20g, which was chosen to provide 
continuous dosing over the early phase of sepsis during the time period when high levels of 
plasma CFH are present. Importantly, there was no signal for harm from Acetaminophen in the 
HEAT trial in terms of liver or other organ dysfunction or mortality. In summary, the HEAT trial 
does not provide any information about the efficacy of extended dosing of intravenous 
Acetaminophen in the target population that is of interest in ASTER and is relevant primarily 
with regards to safety of Acetaminophen in critical illness.  
We will use the FDA-approved dose and frequency 
of intravenous Acetaminophen (1g every 6h or 
4g/day, total dose 20 g; if weight < 50kg then dose 
reduced to 15 mg/kg every 6h). There are extensive 
safety data for Acetaminophen in healthy subjects 
receiving this dose and frequency enterally [41, 42]. 
Specifically, at this dose/frequency: (1) markers of 
liver injury are rarely elevated; (2) liver injury has 
not been observed until after 7 days of 
Acetaminophen use; and (3) any elevation of liver 
injury markers resolved after stopping the drug [41, 
43]. Compared to enteral Acetaminophen, IV 
administration reduces initial hepatic exposure by 
approximately 2-fold through lack of first-pass 
metabolism [44, 45]. This dose of 1g IV every 6 
hours was safe in 346 critically ill patients in the 
Acetaminophen arm in the HEAT trial with some patients receiving Acetaminophen at this dose 
for as long as 3 weeks [40]. In ASTER, the proposed duration of treatment is 5 days, rather 
than the 3 days studied in ACROSS. Treatment duration has been extended because our 
preliminary studies in critically ill patients enrolled in an ICU feeding trial [46], showed that high 
levels of plasma CFH persisted at 6 days (Figure 8). Acetaminophen should continue to be 
protective as long as the oxidative stimulus (CFH) is present; however, we elected to limit 
Acetaminophen treatment to 5 days to minimize any risk of hepatic injury.   
3.4 Summary  
Sepsis is common, associated with organ failure, high mortality, and currently without effective 
therapeutic interventions directed at the underlying pathophysiology. Acute lung injury (ARDS), 
acute kidney injury and cardiovascular dysfunction are common organ injuries in sepsis, are 
associated with increased mortality, and may be mediated, in part, by the elevated levels of 
CFH and pro-oxidant pathways that are characteristic of sepsis. Acetaminophen, a potent 
inhibitor of CFH-mediated oxidative injury, improves lung and renal function in pre-clinical 
models and seems to be potentially beneficial in humans with hemoprotein-mediated diseases, 
including in critically ill adults with sepsis. Vitamin C also has hemoprotein reductant and 
Figure 8 .  Comparison of plasma CFH  
levels in the ACROSS study (days 0,1,2,3)  
and the Vanderbilt phase 2 EDEN study  
( days 0, 6), a trial of different caloric intake  
goals in critically ill patients  


  
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antioxidant effects [28], in addition to cytoprotective and anti-inflammatory effects.  In a phase 
2b clinical trial, Vitamin C improved survival in patients with ARDS, but further study in patients 
with sepsis is needed.   
ASTER is an efficient platform for conducting two phase 2b trials with a shared placebo group in 
which we will assess the potential impact of Acetaminophen and Vitamin C on reducing organ 
injury in a population of patients with sepsis who have been prognostically enriched with 
evidence of either cardiovascular dysfunction or respiratory insufficiency (or both). The trial will 
provide information on both the primary and secondary endpoints, including the biological 
endpoints, that will guide decisions in the future to proceed to larger phase 3 trials that would be 
focused on 90-day mortality. In addition, the results of this trial will establish whether a point-of-
care measurement of the presence of cell-free hemoglobin should be used as a predictive 
enrichment strategy for selecting patients for future clinical trials of Acetaminophen and Vitamin 
C. Also, it is important to note that both intravenous Acetaminophen and Vitamin C are readily 
available and inexpensive, increasing the potential impact of this trial.    
4. Trial Design  
4.1 Study Description  
Prospective multi-center phase 2b randomized placebo-controlled double-blinded interventional 
trials of two different pharmacologic therapies (intravenous Vitamin C or intravenous 
Acetaminophen) for patients with sepsis-induced hypotension or respiratory failure. Patients 
greater than or equal to 18 years old with known or suspected infection with associated 
hemodynamic or acute respiratory dysfunction will be eligible for enrollment.    
Only subjects who are admitted or planned to be admitted to an intensive care unit (ICU) will be 
enrolled. Study drug administration will occur for a total of 120 hours, unless one of the following 
occurs:   
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
5. New AST or ALT elevation greater than 10 times the upper limit of normal level (in the 
APAP-Active/APAP-Placebo group only).  
A total of 900 participants who meet all of the inclusion criteria and none of the exclusion criteria 
will be randomized in a 2:1:2:1 fashion (APAP-Active:APAP-Placebo:Vitamin C-Active:Vitamin 
C-Placebo) into the four treatment arms of the study.  The four treatment arms are: APAP-
Active (300 patients), APAP-Placebo (150 patients), Vitamin C-Active (300 patients), Vitamin C-
Placebo (150 patients). Enrollment will occur over approximately 15 months.  


  
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The study interventions are:  
  
 
 
 
 
 
 
The safety and efficacy will be evaluated through repeated laboratory assessment, clinical 
assessments and biomarker measurements during the entire study. Adverse events and clinical 
outcomes, including need for assisted ventilation, vasopressors, renal replacement therapy, ICU 
length of stay, and hospital length of stay, will be assessed through 28 days or hospital 
discharge, whichever occurs first. Hospital mortality and all cause mortality at 90 days will also 
be assessed.    
4.2 Randomization  
After informed consent and completion of screening, we will randomize each eligible participant 
using a web-based randomization system. Designated research team members will have 
permission to randomize participants. At randomization, each subject will be assigned a unique 
randomization ID number that the site pharmacy can use to determine study arm assignment to 
Acetaminophen-Active, Acetaminophen-Placebo, Vitamin C-Active or Vitamin C-Placebo  
3. Vitamin C-Active given intravenously at the dose of 50 mg/kg every six hours (to 
a maximum dose of 9 grams every six hours) for 5 days (20 doses) OR 
1. APAP-Active given intravenously at the dose of 1 gram (or 15 mg/kg if patient 
weighs < 50 kg) every six hours for 5 days (20 doses) OR 
2. APAP-Placebo (identical appearing 5% dextrose solution) infused every six hours 
for 5  days (20 doses) OR 
4. Vitamin C-Placebo  (identical appearing 5% dextrose solution) infused every six 
hours for 5 days (20 doses)   


  
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(research staff, clinical staff, patient and LAR will be notified if they were randomized to the 
APAP-Active/APAP-Placebo or Vitamin C-Active/Vitamin C-Placebo arm of the study). The CCC 
will send a confirming email to the randomizer containing the randomization ID number. We will 
stratify randomization by enrolling hospital.  
4.3 Primary Efficacy Variable 
Primary efficacy variables will be evaluated comparing Acetaminophen vs. pooled placebo and 
Vitamin C vs. pooled placebo in separate analyses:  
▪ 
Days alive and free of assisted ventilation, vasopressors and new renal replacement 
therapy between either treatment arm and the pooled placebo groups over the first 28 
days (need to be free of all three components to qualify for a day alive and free from 
organ failures).  A patient could be free of all organ failures from day 10 through day 22 
and then die on day 22 and would receive 12 days of credit for this primary outcome. 
The components of this outcome (days free of dialysis, assisted ventilation, and 
vasopressors in the overall cohort and in survivors, and 28-day hospital mortality) will 
also be reported.    
▪ 
This four-component primary outcome was chosen as preliminary data suggest the 
study interventions have the potential to reduce mortality and favorably impact at least 
two of the three organ failure support outcomes for each intervention, thus increasing the 
sensitivity of this outcome for benefit in a Phase 2b trial designed primarily to assess 
safety and proof of concept [47, 48].  
4.4 Secondary Clinical Efficacy Variables 
The ventilator, vasopressor, new renal replacement, and hospital “free days” endpoints below 
are traditional Sepsis and ARDS Phase 2 and Phase 3 composite outcomes that assign a value 
of zero free days for participants dying on or before study day 28 to control for the competing 
risk of death as described in Section 8.3 of the protocol.  These traditional outcomes are 
included in this Phase 2b trial to search for additional evidence of efficacy that could support a 
decision to progress to Phase 3 and will allow for comparison to other sepsis and ARDS trials 
using these endpoints, including all other PETAL studies.   
1. 28-day ventilator-free days  
2. 28-day vasopressor-free days  
3. 28-day new renal replacement-free days  
4. 28-day hospital mortality   
5. 28-day ICU free-days  
6. 28-day hospital-free days to discharge home  
7. 28-day duration of ICU stay in survivors and non-survivors  
8. Initiation of assisted ventilation to day 28  
9. Initiation of renal replacement therapy to day 28  


  
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10. Change in organ-specific Sepsis-related Organ Failure Assessment (SOFA) scores 
between enrollment and study day 7 (Appendix B)  
11. 90-day hospital mortality (includes death at any healthcare facility that is not the 
equivalent of home for a patient)  
12. Development of ARDS within 7 days of randomization  
13. Change in serum creatinine from enrollment to discharge, death, initiation of dialysis 
or 28 days, whichever occurs first   
14. Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline, need for new renal replacement therapy, or death  
15. Change in the Radiographic Assessment of Lung Edema (RALE) score from 
enrollment to study day 3 in patients who are receiving assisted ventilation or high 
flow nasal oxygen at baseline.  
16. 90-day all-cause mortality   
17. Renal stones diagnosed between randomization and study day 90 in the Vitamin C-
Active/Vitamin C-Placebo arm  
NOTE: Please see section 8.3 for description of how these variables are defined and calculated.  
4.5 Safety Variables 
Safety monitoring and analysis will be performed separately on the APAP-Active/APAP-
Placebo arm and the Vitamin C-Active/Vitamin C-Placebo arms (2 active to 1 placebo 
randomization ratio). The safety variables will include the following:  
1. Allergic or allergic-like reactions such as urticarial drug rash, hyperemia with 
hypotension, and anaphylaxis   
2. AST and ALT measured on study days 0, 2-5, and 7 (day 7 measurement can be  1 
day)  in the APAP-Active/APAP-Placebo arm  
3. The trough plasma levels measured just prior to a study dose of Acetaminophen on 
Day 2 (subject must have received at least 5 doses of study drug before the day 2 
samples are obtained) in the first 100 patients randomized to the APAP-
Active/APAP-Placebo arm will be reported to the DSMB to determine if they exceed 
the level of 20 micrograms/mL, the potential level for hepatotoxicity.  
4. Administration of fluid bolus, new use of vasopressor, or increased dose of 
vasopressor within 120 minutes of study drug infusion in the APAP-Active/APAP-
Placebo arm.  
5. Incidence of reported adverse events  
4.6 Biological Endpoints 
Change in plasma IL-6, angiopoietin-2, cell-free DNA, Syndecan-1, and cell-free hemoglobin 
between enrollment (baseline) and Days 2 and 3.  


  
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5. Treatments Arms and Study Drug Administration  
All study drug will be administered by the bedside nurse to the patient through an indwelling 
peripheral or central venous access line.  Study drug will be held if the patient loses all 
intravenous access and will be restarted as soon as intravenous access is restored.  Missed 
doses will be recorded but will not be added on to the end of the study.  If the patient is 
discharged from the ICU prior to day 5 or 20 doses, study drug will be stopped. A list of drug 
incompatibilities for Vitamin C and Acetaminophen will be provided to each clinical site.   
5.1 Acetaminophen-Active Arm  
 
 Patients randomized to Acetaminophen will receive Acetaminophen (hospital pharmacy stock) 
intravenously at the dose of 1 gram (or 15 mg/kg if patient < 50kg) every 6 hours for a maximum 
of 5 days (20 doses). Intravenous Acetaminophen was chosen over enteral Acetaminophen to 
insure adequate drug delivery in critically ill patients. In the ACROSS trial of enteral 
Acetaminophen (1 g every 6 hours), the observed serum Acetaminophen levels were at the 
lower limit of the therapeutic window (Figure 9). Intravenous Acetaminophen is expected to 
provide higher peak levels of Acetaminophen 
while avoiding first pass metabolism by the liver. 
Based on the observation that cell-free 
hemoglobin levels remain elevated at 6 days in 
critically ill patients (see Figure 8, section 3.3), 
treatment will be continued for 5 days (for a 
maximum of 20 doses) or until any of the following 
occur:  
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
5. New AST or ALT elevation greater than 10 
times the upper limit of normal level.  
For blinding purposes, Acetaminophen will be 
transferred to a sterile INTRAVIA container 
(Baxter) or its equivalent prior to dispensing by the 
pharmacy. 
   
Figure 9.    Serum  Acetaminophen   levels on  
enrollment and each study day of the ACROSS  
trial of enteral Acetaminophen versus placebo in  
sepsis   
Treatment: IV Acetaminophen-Active  
Dose: 1 gm (or 15 mg/kg if patient is less than 50 kg)  
Frequency: Every 6 hours  
Duration: 5 days (20 doses) or ICU discharge, whichever comes first  


  
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5.2 Acetaminophen-Placebo Arm  
 
Patients randomized to APAP-Placebo arm will receive an identical appearing room temperature 
5% dextrose in water placebo intravenously every 6 hours for a maximum of 5 days (for a 
maximum of 20 doses). A similar placebo has been used in other randomized clinical trials of IV 
Acetaminophen. The APAP-Placebo will be 100 ml of 5% dextrose in water. Patients weighing 
<50 kg will receive a similar weight-adjusted volume to patients in the active treatment arm. 
Treatment will stop at 5 days (20 doses) or until any of the following occur:  
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
5. New AST or ALT elevation greater than 10 times the upper limit of normal level.   
5.3 Vitamin C Arm-Active  
 
Patients randomized to Vitamin C-Active will receive Vitamin C (Ascor® McGuff Pharmaceuticals) 
intravenously at the dose of 50 mg/kg (to a maximum dose of 9 grams) every 6 hours for a 
maximum of 5 days (for a maximum of 20 doses). The volume of Vitamin C or Placebo arms will 
be lower in patients <80 kg and will be matched in their respective placebos (see pharmacy 
manual). Treatment will stop at 5 days (20 doses) or until any of the following occur:  
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
Study drug will be purchased by the PETAL Network from McGuff Pharmaceuticals and will be 
handled and stored according to manufacturer specifications.  
Treatment: IV Acetaminophen-Placebo 
Dose: 5% dextrose in water  
Frequency: Every 6 hours  
Duration: 5 days (20 doses) or ICU discharge, whichever comes first  
Treatment: IV Vitamin C-Active 
Dose: 50 mg/kg to a max dose of 9 grams 
Frequency: Every 6 hours  
Duration: 5 days (20 doses) or ICU discharge, whichever comes first  


  
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5.4 Vitamin C-Placebo Arm  
 
Patients randomized to Vitamin C-Placebo arm will receive an identical appearing 5% dextrose in 
water placebo intravenously every 6 hours for a maximum of 5 days (for a maximum of 20 
doses). A similar placebo has been used in other randomized clinical trials of IV Vitamin C. The 
Vitamin C-Placebo will be administered in an infusion of 50 or 100 ml of 5% dextrose in water 
dependent upon patient weight. Treatment will stop at 5 days (20 doses) or until any of the 
following occur:  
1. Discharged from the study hospital;   
2. Discharged from the ICU;   
3. Withdrawal from study;   
4. Death;   
 
5.5 Drug Interruptions  
Interruptions of study drug should be avoided. If the study drug must be interrupted due to loss 
of intravenous access, it should be restarted as soon as the treatment team re-establishes 
intravenous access.  In cases in which the study drug was interrupted, the missed doses will not 
be added on to the end of the study. If a dose cannot be administered within 3 hours of the 
scheduled time, the dose should be skipped.  Any dose administered outside of the +/- 1-hour 
window or any dose that is skipped will trigger a protocol deviation.  
Any study drug interruptions will be recorded, including the reason for the interruption.    
In the case of a new measurement of an AST or ALT greater than or equal to 10 times the 
upper limit of normal in the acetaminophen/placebo arm or rash or other hypersensitivity 
attributed to the study drug during the study period, the study drug will be stopped and not 
restarted. Elevation of AST or ALT is known to reverse with discontinuation of Acetaminophen 
[40]; in almost all cases simply discontinuing the study drug is appropriate. If study drug is 
discontinued for AST and/or ALT elevation we will repeat AST and ALT at 24 and 48 hours after 
the study drug is discontinued and continue to measure daily until the levels have declined from 
their peak level.    
6. Study Population and Enrollment  
 6.1 Participant Enrollment  
The study is expected to accrue over approximately 15 months. Participants will be recruited  
Treatment: IV Vitamin C-Placebo 
Dose: 5% dextrose in water 
Frequency: Every 6 hours  
Duration: 5 days (20 doses) or ICU discharge, whichever comes first  


  
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from the Emergency Department or Intensive Care Unit at all PETAL Network centers. 
Participants who withdraw or prematurely discontinue study drug will be analyzed by an 
intention to treat analysis. Safety data will continue to be collected up to hospital discharge for 
any participants who are withdrawn from the study.  
6.2 Inclusion Criteria  
1. Age ≥ 18 years  
2. Sepsis defined as:  
a. Clinical evidence of a known or suspected infection and orders written to administer 
antibiotics  
AND  
b. Hypotension as defined by the need for any vasopressor (and 1 liter of fluid already 
administered intravenously for resuscitation) OR respiratory failure defined by 
mechanical ventilation,  BIPAP or CPAP at any level, or greater than or equal to 6 
liters/minute of supplemental oxygen (criterion b must be present at the time of 
randomization)  
3. Admitted to a study site ICU (or intent for the patient to be admitted to a study site ICU) 
within 36 hours of presentation to the study site ED or within 36 hours of presentation to 
any acute care hospital  
6.3 Exclusion Criteria  
1. No consent/inability to obtain consent from the participant or a legally authorized 
representative  
2. Patient unable to be randomized within 36 hours of presentation to the ED or within 36 hours 
of presentation to any acute care hospital  
3. Diagnosis of cirrhosis by medical chart review  
4. Liver transplant recipient  
5. AST or ALT greater than five times the upper limit of normal   
6. Diagnosis of ongoing chronic alcohol use disorder/abuse by chart review; if medical record 
unclear, use Appendix F  
7. Clinical diagnosis of diabetic ketoacidosis or other condition such as profound hypoglycemia 
that requires hourly blood glucose monitoring  
8. Hypersensitivity to Acetaminophen or Vitamin C  
9. Patient, surrogate or physician not committed to full support (Exception: a patient will not be 
excluded if he/she would receive all supportive care except for attempts at resuscitation from 
cardiac arrest)  
10. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used only 
for sleep-disordered breathing   
11. Chronic dialysis 


  
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12. Current active kidney stone  
13. Multiple (>1) episodes of prior kidney stones, known history of oxalate kidney stones or 
oxalate nephropathy  
14. Kidney transplant recipient  
15. Use of home oxygen for chronic cardiopulmonary disease  
16. Moribund patient not expected to survive 24 hours   
17. Underlying malignancy or other condition with estimated life expectancy of less than 1 month  
18. Pregnant woman, woman of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or woman who is breast feeding  
19. Prisoner  
20. Treating team unwilling to enroll because of intended use of Acetaminophen or Vitamin C  
21. Treating team unwilling to use plasma (as opposed to point of care testing) for glucose 
monitoring 
6.4     Reasons for each of the exclusion criteria  
1. No consent/inability to obtain consent – no justification for waiver of informed consent.   
2. Patient unable to be randomized within 36 hours of presentation: limits enrollment to acute 
conditions and presentations.  
3. Diagnosis of cirrhosis – not safe to administer 4 grams of IV Acetaminophen daily in this 
clinical setting  
4. Liver transplant recipients are excluded as the doses of acetaminophen used in this study 
exceed recommended doses for liver transplant patients   
5. AST or ALT greater than five times the upper limit of normal – this is a standard precaution 
to exclude patients with acute or chronic liver disease [38].   
6. Diagnosis of ongoing chronic alcohol use disorder/abuse – these patients may have an 
increased risk of hepatic toxicity from Acetaminophen.   
7. Presence of diabetic ketoacidosis or other condition such as profound hypoglycemia that 
requires hourly blood glucose monitoring – because glucose measurements cannot be 
reliably done with point of care devices at the bedside in the ICU in the presence of high 
Vitamin C levels, the time lag (30-60 min) in obtaining glucose measurements by other 
methods in this trial makes it impractical to include these patients.  
8. Hypersensitivity to Acetaminophen or Vitamin C – self-evident.  
9. ICU staff, legally authorized next of kin or study subject not committed for full support – 
required to assess the potential beneficial effects of Vitamin C or Acetaminophen on the 
primary and secondary outcomes.  
10. Home assisted ventilation (via tracheotomy or noninvasive) except for CPAP/BIPAP used 
only for sleep-disordered breathing – these patients would have ventilator dependent 
respiratory failure.  


  
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11. Chronic dialysis is excluded because one component of the primary end-point is days free of 
dialysis over the first 28 days in the trial. 
12. Current Active kidney stone – oxalate is a known byproduct of ascorbate metabolism and 
might exacerbate current active kidney stone   
13. Multiple (>1) episodes of prior kidney stones, known history of oxalate stones or oxalate 
nephropathy – oxalate is a known byproduct of ascorbate metabolism and might exacerbate 
existing oxalate stones or oxalate nephropathy in the Vitamin C-Active/Vitamin C-placebo 
arm  
14. Kidney transplant patients - would have a higher risk of oxalate precipitation with Vitamin C  
15. Use of home oxygen for chronic cardiopulmonary disease- these patients will already have 
chronic respiratory failure and therefore would not be a good candidate for interventions that  
are designed to reduce the acute effects of sepsis on respiratory failure.   
16. Moribund patient not expected to survive 24 hours – these patients would not fit the goals of 
testing interventions designed to reduce organ failure from sepsis.  
17. Underlying malignancy or other condition with estimated life expectancy of less than 1 month 
- these patients would not match with the secondary outcome of testing the beneficial effects 
of these therapies on 28 day mortality.   
18. Pregnant female, female of childbearing potential without a documented negative urine or 
serum pregnancy test during the current hospitalization, or female who is breast feeding – 
safety not established for using the interventions to be tested in this trial.  
19. Prisoner – not correct to include these patients in clinical trials by ethical standards.  
20. Treating team unwilling to enroll because of intended use of Acetaminophen or Vitamin C – 
open label Acetaminophen and Vitamin C use are prohibited in this trial because the study 
drug dose is the maximum FDA approved dose and would also confound interpretation of 
the results of the trial.  
21. Use of plasma for glucose testing is required for the vitamin C arm. 
6.5 Co-enrollment 
Co-enrollment in other clinical trials will only be allowed after a trial has been approved for co-
enrollment by ASTER study leadership and the PETAL DSMB. Interventions that are 
contraindicated in combination with Vitamin C or acetaminophen are not permitted.  Study 
procedures of the co-enrolling trial must not impose an undue burden on research participants or 
research staff when viewed within the context of ASTER study procedures, including the total 
volume of blood drawn for research and follow-up activities. For blinded trials, the co-enrolling 
trial must agree to confidentially provide unblinded treatment assignments to the respective 
DSMBs and the unblinded Medical Monitors to allow for accurate assessment of serious adverse 
events and unanticipated problems. 
6.6 Informed Consent  
Screening will occur in the Emergency Department and intensive care units daily to identify 
potential candidates for enrollment. Once a potential participant is identified, the patient’s 
primary medical team will be contacted to secure permission to approach the patient or their 
legally authorized representative regarding possible participation in the study.  If the primary 


  
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team agrees, the study coordinator or investigator will meet with the patient and/or legally 
authorized representative and will describe the proposed study protocol in lay terminology.  
Prior to performing any study procedures, an informed consent form (ICF) approved by the 
central IRB will be signed and dated by each study participant or legally authorized 
representative and the study investigator or person informing the subject and obtaining the 
consent. Prior to taking part in the study, the participant or his or her legal representative will 
receive a copy of the signed and dated ICF.    
A special vulnerable population, cognitively impaired patients, will be eligible for enrollment in 
this proposed study. Many septic patients have cognitive impairment as part of their acute 
disease process and others require sedation during their treatment course. In cases where the 
potential participant is cognitively impaired, surrogate consent will be obtained from the 
participant’s legally authorized representative according to local state law. In these cases and if 
the cognitive impairment has resolved prior to hospital discharge, the participant will be 
reconsented at the earliest opportunity and given the option of continuing or stopping their 
participation.    
6.7 Excluded Medications  
Clinicians will be prohibited from administration of Acetaminophen or high-dose Vitamin C 
(either intravenously or enterally) to study participants during the study.  Additionally, 
administration of open label Acetaminophen or Vitamin C at any dose will be recorded in the 
case report forms.  Multivitamins, supplemental electrolytes/micronutrients or enteral or 
parenteral nutrition that contain Vitamin C and are administered as part of routine clinical care 
will be allowed because these contain a relatively small amount of Vitamin C.   
6.8 On-study Fever Management Recommendations  
Use of nonsteroidal anti-inflammatory drugs (NSAID)s or Cyclooxygenase-2 inhibitors to treat 
fever and pain will be discouraged during the 5-day study drug administration and the use of 
these medications for other indications will also be discouraged. No restriction will be placed on 
the use of aspirin in doses of less than or equal to 325 mg per day.   
The use of open-label acetaminophen will not be permitted while receiving study medication. 
Once the course of study drug administration has been completed, open-label acetaminophen 
can be administered at the discretion of the treating clinician. Physical cooling will be used at 
the discretion of the clinical team per local practice preferences. 
This plan will apply to both the Acetaminophen and placebo arms and also the Vitamin C and 
placebo arms.  Fever Management Recommendations are also outlined in the Appendix.  
6.9 Concomitant Medications  
We will record use of non-steroidal anti-inflammatory agents, iodinated contrast, glucocorticoids 
(all preparations including hydrocortisone, methylprednisolone, prednisone, dexamethasone), 
thiamine, remdesivir, baricitinib, tocilizumab. 


  
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6.10 Blood Glucose Strategy for the Vitamin C-Active/Vitamin C-Placebo Arm  
In patients randomized to the Vitamin C-Active/Vitamin C-Placebo arm, blood glucose 
measurements that are ordered by the treating team for monitoring of blood glucose for clinical 
reasons should be made using the central core laboratory; ABG/stat lab devices or point-of-care 
hand held glucometers should NOT be used.  Hand held (Point-Of-Care, POC) glucometers and 
some ABG/stat laboratory devices currently in clinical use can give falsely elevated glucose 
readings in the setting of high plasma ascorbate levels due to ascorbate oxidation at the surface 
of electrochemical strips contained in these devices [49-51]. The spectrophotometric or 
calorimetric based methods used in clinical core laboratories do not have ascorbate interference 
problems. In the ASTER trial, maximal expected plasma ascorbate concentrations are expected 
to range up to 6 millimoles/L, based on pharmacokinetic data previously obtained in patients 
receiving I.V. ascorbate dosed at 50 mg/kilogram body weight every 6 hours [32]. To our 
knowledge, there are no known point of care glucometers or ABG/stat lab devices that have 
been validated to tolerate this concentration of ascorbate.  24 hours following completion of the 
final dose of Vitamin C-Active/Vitamin C-Placebo, blood glucose measurement can resume per 
unit standards.  
6.11 Common Strategies for All Treatment Arms  
6.11.1 Ventilator Procedures  
Low tidal volume ventilation and weaning from mechanical ventilation will follow standard clinical 
protocol for participants meeting criteria for ARDS (Appendix C).  
Low Tidal Volume Ventilation: We will use a simplified version of the ARDS network 6 ml/kg 
PBW lung protective ventilation protocol. If not already being used, a low tidal volume protocol 
for mechanical ventilation must be initiated within two hours of randomization in all patients who 
meet criteria for ARDS.   
Weaning: Since the time at which a patient achieves unassisted ventilation affects some 
secondary endpoints, and because recent evidence-based consensus recommendations have 
identified a best practice for weaning, a weaning strategy will also be controlled by protocol 
rules in accordance with these evidence-based recommendations. This will assure similar 
weaning methods and provide potential benefit to both study groups. This weaning strategy is a 
simplified version of the protocolized weaning strategy used in prior PETAL and ARDS Network 
studies.  
6.11.2   On Study Fluid Management  
Fluid management during shock will be unrestricted. Balanced crystalloid solution (such as 
lactated Ringers, Hartman’s solution, Plasmalyte or Normosol) is preferred, for resuscitation. 
Given the lack of clear evidence identifying a superior approach, the type of fluid administered 
will not be mandated.   
However, in patients not in shock or after shock resolution, a conservative fluid approach will be 
recommended for patients with ARDS. This conservative fluid management approach is a 
simplification of the algorithm utilized in the ARDS Network FACTT study (Appendix D).   


  
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6.12 Criteria for Discontinuation or Withdrawal  
The primary reason for treatment discontinuation will be recorded using the following categories:  
a. Adverse Event: The participant has experienced an adverse event that the investigator 
believes requires early termination because continued participation imposes an 
unnecessary risk to the participant’s health.  
b. Major Protocol Deviation: The participant failed to meet protocol entry criteria or did not 
adhere to protocol requirements, excluding prolonged interruption of study drug.  
c. Voluntary Withdrawal: The participant or legally authorized representative wishes to 
withdraw from the study. The reason for withdrawal, if provided, will also be documented in 
the CRF.  
d. Other: This category includes participants withdrawn by the treating team for a new 
indication for Acetaminophen or Vitamin C use.  
7.0 Study Plan  
7.1 Baseline Assessments  
The following will be collected from information available prior to randomization. If more than 
one value is available in the 24 hours prior to randomization, the value closest to the time of 
randomization will be recorded.    
1. Demographic and Admission Data (including age, sex, race)  
2. Pertinent Medical History and Physical Examination (including components of the 
Charlson co-morbidity score)  
3. Height; gender; measured Body Weight (mBW); calculated predicted body weight 
(PBW); body mass index (BMI)  
4. Location when inclusion criteria met: ED, ICU  
5. Vital signs:  heart rate (beats / min), systemic systolic and diastolic BP (mmHg), body 
temperature (°C), and oxygen saturation with FiO2  
6. Use of vasopressors, corticosteroids, acetaminophen, thiamine, supplemental O2, 
oxygen saturation, invasive or non-invasive/invasive mechanical ventilation and renal 
replacement therapy prior to randomization   
7. Clinical laboratory testing results ( AST, ALT, total bilirubin)  
8. Serum creatinine: 
a Prehospitalization: Most recent outpatient creatinine from 24 hours to 365 days 
prior to admission if available 
b Baseline: Lowest in hospital creatinine prior to randomization  
9. Chest radiograph   
10. ARDS assessment if receiving assisted ventilation  


  
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11. Presumed site of primary infection  
12. SOFA Score: as described in Appendix B  
13. Glasgow Coma Score 
14. Cell free plasma hemoglobin (to be collected and measured in batch at the end of the 
study)  
15. COVID-19 status and date of first positive test if known within 3 weeks of enrollment 
 7.2. Assessments During Protocol Phase (through study day 7)  
1. Daily fluid administration for the first 7 days (truncated for death or ICU discharge)  
2. Daily fluid outputs for the first 7 days (truncated for death or ICU discharge)  
3. Chest radiograph on day 3 (± 1 day) for all patients who were receiving assisted 
ventilation or high flow nasal oxygen at the time of randomization if they are still in the 
study hospital  
4. ARDS assessment through study day 7 (± 1 day). We will determine the presence and 
severity of ARDS for each day of assisted ventilation to day 7 using the following 
approach.  For participants on assisted ventilation with P/F <300 or S/F <315, FiO2 
≥40%, and PEEP ≥5 cm H2O, determine if hypoxemia is valid, acute, and not fully 
explained by CHF or fluid overload. If yes, local investigators will review the first CXR 
(or CT) performed on each ventilated day with valid P/F or S/F <315 (to day 7). If no 
chest imaging studies are present that day, site investigators may review available 
imaging one day before or after to determine if ARDS imaging criteria met. ARDS 
imaging criteria are met if the images are consistent with ARDS (bilateral opacities not 
fully explained by effusions, lobar/lung collapse, or nodules). If equivocal, the reviewing 
investigator will adjudicate with additional investigators.   
5. AST and ALT, and bilirubin on days 1-7 if available.   AST/ALT REQUIRED on days 2-
5, and 7 (day 7 measurement can be 1 day)  in the APAP-Active/APAP-Placebo 
arm  
6. Daily SOFA score through day 7 or hospital discharge   
7. Recording of fluid bolus (500 mL or more), new vasopressor administration, or increase 
of vasopressor dose within 120 minutes of receiving study drug in the APAP-
Active/APAP-Placebo arm.  
8. Administration of the following concomitant medications: non-steroidal anti-inflammatory 
agents, iodinated contrast, glucocorticoids, thiamine, remdesivir, baricitinib, and 
tocilizumab   
7.3. Other Assessments and Data Collection  
1. Vasopressor use over 28 days  
2. Study drug administration 
3. Use of invasive and non-invasive ventilation (except noninvasive ventilation solely for 
sleep disordered breathing) over 28 days  


  
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4. New renal replacement therapy over 28 days  
5. Last creatinine prior to initiation of RRT, death, study hospital discharge, or day 28 
(whichever comes first)  
6. Highest creatinine between randomization and day 28.  
7. Results of microbiologic testing from samples taken from up to 24 hours prior to 
randomization until 72 hours after randomization 
8.  Source of infection  
7.4. Assessments after Hospitalization  
We will perform an assessment using all testing and records available during the hospitalization 
where a site investigator will make an assessment as to whether the hypotension at enrollment 
was likely due to an infection or not.  
7.5. 90-day All-Cause Mortality  
We will contact patients at day 90 to ascertain their survival status.  In the Vitamin C-
Active/Vitamin C-Placebo arm, we will also ascertain the development of renal calculi (kidney 
stone). This will be done by telephone contact with the patient or family members as well as a 
review of medical records.  We will use publicly available data sources and the national death 
index as a final check for patients with whom we are unable to confirm their vital status through 
other means.  
7.6. Specimen Collection  
Plasma: Blood will be collected within 2 hours of randomization (prior to first study drug 
administration) and on days 2 and 3 (approximately 36 mls total).  To minimize hemolysis, blood 
will preferentially be collected through central venous access devices if present. If there is no 
central venous access, then either an arterial or peripheral venous stick can be used, with blood 
drawn slowly to minimize hemolysis. The blood samples will be centrifuged, and the plasma will 
be frozen and stored at a biorepository for future research for the following purposes:  
• 
Biological Endpoint Assays: To fulfill the secondary biological endpoints, the plasma 
samples will be measured for the concentration of plasma IL-6, angiopoietin-2, 
Syndecan-1, and cell-free DNA on enrollment and on days 2 and 3 and analyzed for 
significant changes from baseline in the pooled placebo, Vitamin C and Acetaminophen 
groups.  
• 
Additional Assays: In addition, the plasma samples at baseline and on days 2 and 3 will 
be used to measure cell-free hemoglobin.  
• 
Acetaminophen Levels: Plasma collected as a trough level just prior to a study dose on 
Day 2 (subject must have received at least 5 doses of study drug before the day 2 
samples are obtained) will be used to measure Acetaminophen levels in the first 100 
patients randomized to the Acetaminophen arm.  Measuring Acetaminophen levels in 
critically ill patients with sepsis will address a knowledge gap in the literature and 
contribute to interpretation of the results of the trial and also provide data that the DSMB 
can review after 100 patients are enrolled in the Acetaminophen arm. For the first 100 
patients in the acetaminophen arm, the blood drawn on day 2 (between 8 am and 2 pm) 


  
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will be timed to occur 30 minutes before the next study drug administration (for a trough 
level of acetaminophen).   
• 
Samples for Future Research: Plasma that has not been used for these planned 
assays will be stored in a biorepository for future research funded by ancillary studies.  
Whole Blood for Genetics: When consent for genetic testing is specifically obtained, additional 
whole blood for future RNA and DNA studies will be collected at baseline and on day 3. Total 
blood volume for the genetic samples is approximately 8.5 ml/day, or 17mls total. Total overall 
blood volume collected (plasma and DNA/RNA) is approximately 53 ml.   
Urine: Urine will be collected within 2 hours of randomization and on days 2 and 3 (if patient still 
in the ICU), frozen and stored along with plasma. Urine samples will be saved for ancillary 
studies.  
Storage and Shipment: All samples collected in this trial will be identified by a coded number 
during shipment and storage at the central biorepository. 
8. Statistical Methods  
8.1 Primary Endpoint  
The primary endpoint of this trial is to evaluate the effects of intravenous Acetaminophen 
compared to pooled placebo and intravenous Vitamin C compared to pooled placebo on the 
number of days alive and free of assisted ventilation, vasopressors and new renal replacement 
therapy between either treatment arm and the pooled placebo group over the first 28 days.   
Note that the primary outcome components to be reported will be:  
1. 28-day mortality prior to discharge home   
2. Days free of assisted ventilation to day 28 in the overall cohort and survivors  
3. Days free of new RRT to day 28 in the overall cohort and survivors  
4. Days free of vasopressors to day 28 in in the overall cohort and survivors  
The primary safety objective is to determine the effect of intravenous Acetaminophen on markers 
of liver injury (AST and ALT) measured on days 2-5, and 7 (day 7 measurement can be 1 day).  
8.2   Sample Size  
The primary study outcome is 28-day organ support free days (alive and without assisted 
ventilation, new renal replacement and vasopressors). Using clinical trial data from the PETAL 
Network, the estimated standard deviation of 28-day organ support free days is 10 days. With a 
sample size of 300 per active treatment group, and 150 per placebo treatment group, a two 
sample t-test will have 85% power at a two-sided alpha level of 0.05 to detect a significant 
difference between active treatment and pooled placebo if the true mean difference is 2.5 
support free days. We make no multiple comparison adjustment to account for simultaneously 
testing two active treatments since each treatment addresses a distinct scientific question [52].  


  
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8.3 Secondary endpoints  
We will compare the efficacy of intravenous Acetaminophen to pooled placebo and intravenous 
Vitamin C to pooled placebo for efficacy endpoints.  We will compare the safety of APAP to 
APAP-Placebo and Vitamin C to Vitamin C-Placebo for safety endpoints:  
• 
28-day ventilator-free days: VFDs depend on both duration of ventilation and mortality 
through study day 28. In participants who survive 28 days, VFD is defined as 28 minus 
days of invasive or noninvasive ventilation to day 28.  Duration of ventilation is counted 
from the first study day of assisted breathing through the last day of assisted breathing 
provided the last day is prior to day 28. For participants discharged with assisted 
ventilation (e.g., to LTAC facility) prior to day 28, a phone call will be required to assess 
ventilator and vital status at day 28. Participants discharged prior to day 28 on 
unassisted breathing will be assumed to remain on unassisted breathing through day 28. 
Isolated periods of ventilation briefer than 24 hours for surgical procedures and 
ventilation solely for sleep disordered breathing do not count towards duration of 
ventilation. In participants who never require assisted breathing, duration of ventilation is 
zero. Participants who do not survive 28 days will be assigned zero VFD.  
• 
28-day vasopressor-free days: Vasopressor free days to day 28 are defined as the 
number of calendar days between randomization and 28 days later that the patient is 
alive and without the use of vasopressor therapy.  Patients who die prior to day 28 and 
those who receive vasopressor therapy for the entire first 28 days are assigned zero 
vasopressor free days.    
• 
28-day new renal replacement-free days: Renal replacement free days to day 28 are 
defined as the number of calendar days between randomization and 28 days later that 
the patient is alive and without renal replacement therapy. We also follow the “last off” 
method. Patients who died prior to day 28 and those who receive renal replacement 
therapy for the entire first 28 days are assigned zero renal replacement free days.  
• 
28-day hospital mortality  
• 
28-day ICU free days:  ICU free days to day 28 are defined as the number of days 
spent alive and out of the ICU to day 28.   
• 
28-day hospital free days to discharge home: Hospital free days to day 28 are 
defined as 28 days minus the number of days from randomization to discharge home. If 
a patient has not been discharged home prior to study day 28 or dies prior to day 28, 
hospital free days will be zero. Patients transferred to another hospital or other health 
care facility will be followed to day 28 to assess this endpoint.  
• 
Duration of ICU stay in survivors and non-survivors: The total number of days spent 
in the ICU until hospital discharge or death during the first 28 days. If a patient is 
discharged alive from the study hospital we assume they are no longer in the ICU.  
• 
Initiation of assisted ventilation to day 28:  Any patient who received assisted 
ventilation in the first 28 days meets this endpoint. If a patient leaves the hospital without 
initiation of assisted ventilation, we assume they never started.  
• 
Initiation of new renal replacement therapy to day 28.  
o Patients who receive (new) renal replacement therapy through day 28 will meet 
this endpoint.  


  
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• 
Change in organ-specific Sepsis-related Organ Failure Assessment (SOFA) scores 
between enrollment and day 7: We will calculate the SOFA score upon enrollment and 
at day 7 using clinically available data. If a value is not available at baseline, it will be 
assumed to be normal. At the day 7 assessment, if a value is missing then we will carry 
forward the closest previously known value. The GCS component of the SOFA score will 
be omitted.  (See Appendix B)  
• 
90-day hospital mortality (any hospital)  
• 
Development of ARDS within 7 days: We will determine the presence and severity of 
ARDS for each day of assisted ventilation to day 7. For participants on assisted  
ventilation with P/F <300 or imputed P/F <300, FiO2 ≥40%, and PEEP ≥5 cm H2O, 
determine if hypoxemia is valid, acute, and not fully explained by CHF or fluid overload. 
If yes, local investigators will review the first CXR (or CT) performed on each ventilated 
day with valid P/F or imputed P/F <300 (to day 7). If no chest imaging studies are 
present that day, site investigators may review available imaging one day before or after 
to determine if ARDS imaging criteria are met. ARDS imaging criteria are met if the 
images are consistent with ARDS (bilateral opacities not fully explained by effusions, 
lobar/lung collapse, or nodules). If equivocal, the reviewing investigator will adjudicate 
with additional investigators.  
• 
Change in serum creatinine from enrollment to discharge, death, initiation of dialysis, or 
day 28, whichever occurs first.  
• 
Major Adverse Kidney Events at 28 days (MAKE28): persistent increase in serum 
creatinine by 200% from baseline (defined as most recent outpatient creatinine from 24 
hours to 365 days prior to admission if available: if not available, lowest in hospital 
creatinine prior to randomization), need for renal replacement therapy, or death  
• 
Change in Radiographic Assessment of Lung Edema (RALE) score from enrollment to 
study day 3 in patients who are receiving assisted ventilation or high flow nasal oxygen 
at the time of study randomization  
• 
The safety of intravenous Acetaminophen compared to APAP-Placebo and intravenous 
Vitamin C compared to Vitamin C-Placebo as measured by daily serum AST, ALT, blood 
pressure following study drug administration (for the APAP-Active/APAP-Placebo arm), 
and incidence of reported adverse events (both arms)  
• 
Change in plasma IL-6, plasma IL-8, plasma angiopoietin-2, plasma cell-free 
hemoglobin, syndecan-1, and plasma cell-free DNA from enrollment to days 2 and 3  
• 
90 day all-cause mortality: We will contact patients at day 90 to ascertain their survival 
status. This will be done by telephone contact with the patient or family members as well 
as a review of medical records and publicly available data sources. We will use the 
national death index as a final check for patients with whom we are unable to confirm 
their vital status through other means.  
• 
Renal calculi diagnosed between randomization and study day 90 in patients in the 
Vitamin C-Active/Vitamin C-Placebo group.  This will be ascertained by medical record 
review or during the phone contact with the patient or family members at day 90.  


  
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8.4 Safety Endpoints  
AST and ALT will be measured on days 2, 3, 4,  5, and 7 after study enrollment in the APAP-
Active-APAP-Placebo arm. We will also collect data daily on potential adverse reactions to 
Acetaminophen and Vitamin C, such as rash and hypersensitivity.  If either are present and 
thought to be due to the study drug, the study drug will be stopped.  
8.5 Predictive Biomarker  Analysis  
We will  evaluate the predictive value of baseline plasma CFH for identifying patients treated 
with either Acetaminophen or Vitamin C who have an improvement in the primary endpoint, the 
three components of the primary endpoint, 28-day mortality or improvement in renal function as 
assessed by MAKE28. 
8.6 Statistical Analysis Plan  
The principal analyses will be intention-to-treat (based upon randomization assignment). A 
comprehensive statistical analysis plan will be written prior to the un-blinding of the primary and 
secondary efficacy results.  
8.6.1 Sub-Group by Treatment Interaction 
We will do a sub-group by treatment interaction analysis comparing outcomes in acute COVID-
19, defined as a positive diagnostic test for SARS CoV-2 in the prior three weeks and 
pneumonia as primary site of infection, and COVID-19 negative patients for the primary and 
secondary efficacy endpoints. The analysis will include a tabulation of the treatment effect in 
COVID-19 positive patients, the treatment effect in COVID-19 negative patients, and the 
difference between the treatment effect in these two sub-groups. 
8.7 Interim Analysis  
The DSMB will have the ability to analyze all safety data, including AST/ALT, at any time during 
the study and stop the study indefinitely for safety reasons. The safety report will include a 
tabulation of all baseline demographics and physiologic variables as well as on-study variables, 
including SOFA scores, and all adverse events by randomized groups. For monitoring safety, 
data on 28-day mortality will be included with 95% confidence intervals for any differences 
between pooled placebo and the two treatment groups. In addition, a formal safety analysis will 
be done after 450 patients are accrued. This report will include an analysis of acetaminophen 
levels.  Trough levels greater than 20 mcg/mL will be evaluated for any relationship to AST/ALT 
levels.  Investigators will recommend protocol modification as needed to the DSMB as part of 
the safety review.   
Based on an interim analysis of the primary efficacy variable, this trial may stop for efficacy or 
futility of APAP relative to pooled placebo or Vitamin C infusion relative to pooled placebo. The 
stopping rules are based on two stage Haybittle-Peto stopping boundaries [53, 54] with Z=3.4 
for early efficacy stopping. We plan one interim look at approximately one half of total 
enrollment and a final look at full enrollment.   
Table 1 below presents the stopping boundaries as a function of the mean difference in the 
primary efficacy variable favoring active treatment and the one-sided p-value favoring active 


  
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treatment. Tables 2 and 3 present the probability of stopping at stage one under the null and 
alternative hypotheses respectively. As can be seen in Table 2 the probability of stopping early 
for futility under the null is 10%.  Table 3 shows that the probability of stopping early for efficacy 
under the alternative is also 10%.  
Table 1. Stopping Boundaries  
n 
Futility Bound  
Mean Difference 
Efficacy Bound  
Mean Difference 
Futility Bound   
P-value 
Efficacy Bound  
 P-value 
~450 
-1.48066 
3.93066 
0.89986 
0.0003369 
900 
1.60312 
1.60312 
0.02493 
0.02493 
 
Table 2. Stopping probabilities given the null hypothesis 
Stage  
Futility  
Efficacy  
1  
0.10014  
0.00034  
 
Table 3. Stopping probabilities given the alternative hypothesis 
Stage  
Futility  
Efficacy  
1  
0.00034  
0.10014  
  
If one arm of the trial is stopped by the DSMB at the study midpoint (n=150 in each active arm; 
75 in each matching placebo arms), subsequent patients would be randomized one to one to 
the remaining active and matching placebo arms (additional 150 in each arm).  At study close, 
the efficacy analysis dataset for the completed study will consist of all patients randomized to 
active drug (n=300) compared to all placebo patients (n=300, includes 225 in the matching 
placebo arm and 75 in the unmatched arm from the first period enrolled before that arm 
stopped).  The efficacy comparison for the arm that stopped early will be all patients randomized 
to active drug (n=150) and all placebo patients randomized prior to study midpoint (n=150).  
Safety analyses for both studies would compare the active arms versus the matching placebo 
arms, that is, 300 versus 225 and 150 versus 75 for the study that continued to completion and 
stopped early, respectively.  
9.0 Risk Assessment  
This study involves randomization to one of four groups: APAP-Active, APAP-Placebo, Vitamin 
C-Active, or Vitamin C-Placebo within 36 hours of meeting inclusion. Patients will all be closely 
monitored in an ED or ICU setting. This will allow for prompt treatment of any suspected 
adverse events, including liver injury. Adverse events resulting in discontinuation of study drug 
will be reported to the DSMB as noted in section 11.1 and Appendix E.    
9.1 Potential Risks to Subjects  
Risk of Vitamin C administration:  High dose Vitamin C therapy is a powerful anti-oxidant and 
micronutrient and a safe therapy in normal subjects and in critically ill patients. Muhlhofer and 
colleagues [30] examined for the presence of adverse effects following intravenous infusion of 
7500 mg Vitamin C daily for 6 days in normal subjects (n=6). No abnormalities in laboratory 
analyses were found. Nathens et al infused high dose Vitamin C (1500 mg three times daily) 


  
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into critically ill surgical patients for 28 days and found no serious adverse events [29]. Hoffer et 
al infused dosages as high as 1.5 grams/kg body weight three times weekly into patients with 
advanced cancer [55]. Adverse events were infrequent consisting of nausea, vomiting, 
dizziness, and headache.  A systematic review conducted by Biesalski [56]  on the safety of the 
long term low dose parenteral administration of Vitamin C in patients on hemodialysis revealed 
it to be safe with frequent monitoring of oxalate following dialysis. In a safety study conducted at 
VCU, testing the safety of infusing high doses of Vitamin C in patients with sepsis, in which 
approximately 8 of 24 subjects were receiving dialysis, no adverse events occurred that were 
related to the ascorbic acid. Finally, in the recently completed CITRIS-ALI study of high dose 
Vitamin C versus placebo in patients with ARDS, no adverse events related to Vitamin C were 
identified. We will exclude patients with active kidney stone, recurrent kidney stones, a history of 
oxalate stones or oxalate nephropathy or patients who have undergone a renal transplantation.   
Risk of Acetaminophen Administration: Serum AST and ALT will be monitored as part of the 
study on study days 0 and days 2-5.  New measured values of AST or ALT greater than or 
equal to 10 times the upper limit of normal on any measurement in the APAP-Active/APAP-
Placebo group will prompt discontinuation of the study drug, which will not be restarted. 
Elevations in AST and ALT are common as a result of severe sepsis regardless of an 
interventional trial, are rarely associated with this dose and schedule of Acetaminophen, and do 
not require any changes in therapy other than stopping of the study drug. Therefore, the risk of 
AST or ALT elevation will be included in the informed consent and AST or ALT elevations 
greater than 10 times the upper limit of normal will not be considered serious adverse events.  
Reporting of all adverse events to the DSMB will be done in a blinded fashion. In addition to 
reporting these events to the DSMB, all events will also be included in the annual review by the 
IRB.    
Hypotension is a rare but described occurrence with intravenous Acetaminophen administration 
(frequency of 1% per manufacturer’s package insert) that appears to occur predominantly in 
patients with fever. We will monitor the need for a fluid bolus (500 ml or more), the need for a 
new vasopressor, or an increase in the vasopressor dose in the 120 minutes following 
administration of the study drug. Risk of hypotension will be included in the informed consent.   
Risk of Placebo Administration: Minimal risk because the volume of administration of 5% 
Dextrose in water is only 100 ml every 6 hours for 120 hours.  
Risks of Blood Draws: All patients will have blood drawn for research purposes. Many patients 
will have invasive lines placed for clinical purposes, where the risk of blood draws is extremely 
low, as blood can usually be easily obtained from these lines. In cases where an invasive line is 
not present, the risks of drawing blood are uncommon and include bleeding and bruising. 
Commonly, drawing blood is painful, and rarely, drawing blood can lead to infections at the site 
of the blood draw.  
9.2  Risk of Death  
It is possible that one treatment arm may lead to more deaths in comparison to placebo; 
mortality will be reported to the DSMB at the interim and final analyses.  


  
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9.3  Minimization of Risk  
Federal regulations at 45 CFR 46.111(a)(1) require that risks to subjects are minimized by using 
procedures which are consistent with sound research design. There are several elements of 
study design inherent in the present protocol that meets this human subject protection 
requirement. The DSMB will be reviewing data as outlined above and will examine not only 
efficacy but safety (inclusive of mortality) and will reserve the right to halt the study at any time.   
9.4 Potential Benefits  
Study subjects may or may not receive any direct benefits from their participation in this study. It 
is unclear whether Vitamin C or Acetaminophen will provide benefit.  Thus, the optimal 
treatment with Vitamin C or Acetaminophen is undefined at this point.   
9.5 Risks in Relation to Anticipated Benefits  
Federal regulations at 45 CFR 46.111 (a)(2) require that “the risks to subjects are reasonable in 
relation to anticipated benefits, if any, to subjects, and the importance of the knowledge that 
may reasonably be expected to result.”  Based on the preceding assessment of risks and 
potential benefits, the risks to subjects are reasonable in relation to anticipated benefits.    
10. Human Subjects  
Each study participant or a LAR must sign and date an informed consent form. Institutional 
review board approval will be required before any subject is entered into the study. PETAL will 
use a central IRB.  
 10.1 Selection of Subjects  
Federal regulations at 45 CFR 46(a)(3) require the equitable selection of subjects. The EDs and 
the ICUs will be screened to determine if any patient meets inclusion and exclusion criteria. 
Data that have been collected as part of the routine management of the subject will be reviewed 
to determine eligibility. No protocol-specific tests or procedures will be performed as part of the 
screening process. If any subjects meet criteria for study enrollment, then the attending 
physician will be asked for permission to approach the patient or his/her LAR for informed 
consent. Study exclusion criteria neither unjustly exclude classes of individuals from 
participation in the research nor unjustly include classes of individuals from participation in the 
research. Hence, the recruitment of subjects conforms to the principle of distributive justice.   
 10.2   Justification for Including Vulnerable Subjects  
The present research aims to investigate the safety and efficacy of a type of treatment for 
patients with sepsis induced hypotension or acute respiratory failure. Due to the nature of sepsis 
induced hypotension and respiratory failure, most of these patients will have impaired decision-
making capabilities. This study cannot be conducted if limited to enrolling only those subjects 
with retained decision-making capacity. Hence, subjects recruited for this trial are not being 
unfairly burdened with involvement in this research simply because they are easily available.  


  
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10.3 Informed Consent  
Federal regulations 45 CFR 46.111(a)(5) require that informed consent will be sought from each 
prospective subject or the subject’s LAR. The one obtaining consent is responsible for ensuring 
that the patient and/or LAR understands the risks and benefits of participating in the study, and 
answering any questions the LAR may have throughout the study and sharing any new 
information in a timely manner that may be relevant to the LAR’s willingness to permit the 
subject’s continued participation in the trial. The consenter will make every effort to minimize 
coercion. All study participants or their LARs will be informed of the objectives of the study and 
the potential risks. The informed consent document will be used to explain the risks and benefits 
of study participation to the LAR in simple terms before the patient is entered into the study, and 
to document that the LAR is satisfied with his or her understanding of the risks and benefits of 
participating in the study and desires to participate in the study.  The investigator is responsible 
for ensuring that informed consent is given by each patient or LAR. This includes obtaining the 
appropriate signatures and dates on the informed consent document prior to the performance of 
any protocol procedures including administration of study agent.  
10.3.1 Process of Obtaining Informed Consent 
Informed consent will be obtained from the patient or from a surrogate decision maker if the 
patient lacks decision-making capacity.   In some instances, bringing a paper consent form and 
pen to the bedside of a patient with known or suspected COVID-19 and then taking these out of 
the room would violate infection control principles and policies.  Given that many of the potential 
ASTER patients will not yet have been ruled out for COVID-19 at the time of consent, the use of 
“no-touch” consent methods will be allowed.  Additionally, the electronic consent platform 
facilitates documentation of informed consent from legal authorized representatives who are not 
present in the hospital at the time that the LAR is approached for informed consent. 
Below, we outline three examples of no-touch consent procedures that may be used: (a) a paper-
based approach (method 1); (b) a second paper-based approach (method 2); and (c) an 
electronic/e-consent approach (method 3). 
 
 Method 1 Paper-based approach:  
1. The informed consent document is delivered to the patient or LAR. 
a. If the patient or LAR is on-site, the informed consent document many be delivered to 
the patient or LAR either by research staff or by clinical staff 
b. If the LAR is off-site, the informed consent document may be emailed, faxed, or 
otherwise electronically transferred to the LAR (method dictated by institutional policy) 
2. Research staff discuss the informed consent document with the patient or LAR either in-
person or by telephone or videophone.  This step confirms subject/LAR identity. 
3. If the patient or LAR decides to consent to participate, the patient or LAR signs and dates 
the paper copy of the informed consent document. 
4. A photograph is taken of the signature page of the informed consent document and 
uploaded into the electronic database (e.g. REDCap).  
a. If using the patient’s device (such as a patient’s personal cellular phone), a survey link 
can be sent to their device to allow direct upload of the image into the electronic 
database (e.g. REDCap). 


  
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b. If using a staff device, it must be approved to store PHI by the local institution. In that 
case, research personnel can take a photograph of the signature page of the informed 
consent document either directly or through the window or glass door leading into the 
patient’s room.  The photograph can then be uploaded into the electronic database.  If 
a staff device is taken into the patient’s room to take a photograph it must be able to 
be disinfected according to local institutional practices.  
5. Research staff (and witness if applicable based on local requirements) provide signatures 
within the electronic database (e.g. REDCap) confirming their participation in the informed 
consent process. 
6. The patient or LAR retains the paper consent document. The image of the signature page 
may be printed and bundled with a copy of the blank informed consent document for 
research records. 
 
Method 2 Paper-based approach: 
A photograph of the signed ICF can be transmitted to trial staff. 
1. An unsigned ICF is provided to the patient by a person who has entered the room. 
2. The investigator/designee arranges a telephone call or videoconference call with the patient 
(and, if desired and feasible, additional individuals requested by the patient [e.g., next of kin]). 
3. To ensure the patients are approached in a consistent fashion, a standard process should be 
used that will accomplish the following:  
a) Identification of who is on the call. 
b) Review of the ICF with the patient by the investigator/designee and response to any 
questions the patient may have. 
c) Verbal confirmation by the patient that their questions have been answered, that they 
would like to participate in the trial, and that they have signed and dated the ICF that is in 
their possession. 
4. The patient (or an individual in the room) takes a photograph of the signed and dated ICF and 
sends it to the investigator/designee. 
5. Research staff and LAR/patient provide signatures confirming their participation in the 
informed consent process. 
6. The patient or LAR retains the paper consent document. The image of the signature page 
may be printed and bundled with a copy of the blank informed consent documents for 
research records.   
 Method 3 Electronic/e-consent approach: 
1. The electronic informed consent document is opened on a research device or a link for the 
electronic informed consent document is sent to the patient’s or LAR’s device. 
2. Research staff discuss the informed consent document with the patient or LAR either in 
person or by telephone or videophone.  This step confirms subject/LAR identity. 
3. If the patient or LAR decides to consent to participate the patient or LAR signs the electronic 
informed consent document.  This signature may be either: 
a. an actual signature (often tracing a finger on the screen) OR 
b. a username and password specific to the individual signing 
4. Research staff and witness (if required by institutional standards) provide signatures within 
the electronic database (e.g., REDCap) confirming their participation in the informed consent 
process. 


  
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If a hospital device is provided to facilitate electronic or paper-based consent, that device will be 
disinfected according to institutional protocols and removed by research staff or clinical staff 
during the next entry into the patient’s room. 
This approach complies with relevant regulations and sub-regulator guidance at 45 CFR 46.117, 
45 CFR 164.512, 21 CFR 11 Subpart C (11.100–11.300), https://www.hhs.gov/ohrp/regulations-
and-policy/guidance/use-electronic-informed-consent-questions-and-answers/index.html, 
https://www.fda.gov/regulatory-information/search-fda-guidance-documents/informed-consent 
The information for the informed consent discussion will be provided in a formal document (or 
electronic equivalent) that has been approved by the IRB and in a language comprehensible to 
the potential participant, using an interpreter if necessary. The information presented in the 
consent form and by the research staff will detail the nature of the trial and what is expected of 
participants, including any potential risks or benefits of taking part. It will be clearly stated that the 
participant is free to withdraw from the trial at any time for any reason without prejudice to future 
care, and with no obligation to give the reason for withdrawal. Where a patient does not speak 
English, a short-form consent and qualified interpreter will be employed, using similar “no-touch” 
principles.  Use of an interpreter and the interpreter’s identity will be documented on the 
electronic consent. 
 10.4   Continuing Consent  
Subjects for whom consent was initially obtained from a LAR, but who subsequently regain 
decision-making capacity while in the hospital, will be approached for consent for continuing 
participation, including continuance of data acquisition. The consent form signed by the LAR 
should reflect that such consent will be obtained. 
10.5 Withdrawal of Consent  
Patients or the LAR may withdraw or be withdrawn (by the LAR) from the trial at any time 
without prejudice. Data recorded up to the point of withdrawal will be included in the trial 
analysis, unless consent to use their data has also been explicitly withdrawn. If a patient or LAR 
requests termination of study procedures during the treatment period, the procedures will be 
stopped but the patient will continue to be followed up as part of the trial. If a patient or LAR 
withdraws consent during trial treatment, the study procedures will be stopped but permission 
will be sought to access medical records for data related to the trial. If a patient or LAR wishes 
to withdraw from the trial after completion of trial treatment, permission to access medical 
records for trial data will be assumed unless explicitly withdrawn.    
 10.6    Identification of Legally Authorized Representatives  
Many of the patients approached for participation in this research protocol will have limitations of 
decision-making abilities due to their critical illness. Hence, some patients will not be able to 
provide informed consent.  Accordingly, informed consent will be sought from the potential 
subject’s legally authorized representative (LAR).    
Regarding proxy consent, the existing federal research regulations (‘the Common Rule’) states 
at 45 CFR 46.116 that “no investigator may involve a human being as a subject in 
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subject or the subject’s legally authorized representative”; and defines at 45 CFR 46 102 (c) a 
legally authorized representative (LAR) as “an individual or judicial or other body authorized 
under applicable law to consent on behalf of a prospective subject to the subject’s participation 
in the procedures(s) involved in the research.” The Office of Human Research Protections 
(OHRP) defined examples of “applicable law” as being state statutes, regulations, case law, or 
formal opinion of a State Attorney General that addresses the issue of surrogate consent to 
medical procedures. Such “applicable law” could then be considered as empowering the LAR to 
provide consent for subject participation in the research. Interpretation of “applicable law” may 
be state specific and will be addressed by the PETAL central IRB.  
According to a previous President’s Bioethics Committee (National Bioethics Advisory 
Committee (NBAC)), an investigator should accept a relative or friend of the potential subject 
who is recognized as an LAR for purposes of clinical decision making under the law of the state 
where the research takes place [56]. Finally, OHRP has stated in their determination letters that 
a surrogate could serve as a LAR for research decision making if such an individual is 
authorized under applicable state law to provide consent for the “procedures” involved in the 
research study [57].  
10.7    Justification of Surrogate Consent  
According to the Belmont Report, respect for persons incorporates at least two ethical 
convictions; first, that individuals should be treated as autonomous agents, and second, that 
person with diminished autonomy are entitled to protection. One method that serves to protect 
subjects is restrictions on the participation of subjects in research that presents greater than 
minimal risks. Commentators and research ethics commissions have held the view that it is 
permissible to include incapable subjects in greater than minimal risk research as long as there 
is the potential for beneficial effects and that the research presents a balance of risks and 
expected direct benefits similar to that available in the clinical setting [58]. Several U.S. task 
forces have deemed it is permissible to include incapable subjects in research. For example, the 
American College of Physicians’ document allows surrogates to consent to research involving 
incapable subjects only  “if the net additional risks of participation are not substantially greater 
than the risks of standard treatment.” [59]. Finally, NBAC stated that an IRB may approve a 
protocol that presents greater than minimal risk but offers the prospect of direct medical benefits 
to the subject, provided that “the potential subject’s LAR gives permission…” [57]. Consistent 
with the above ethical sensibilities regarding the participation of decisionally incapable subjects 
in research and the previous assessment of risks and benefits in the previous section, the 
present trial presents a balance of risks and potential direct benefits that is similar to that 
available in the clinical setting, with the exception of the additional blood draws.      
 10.8     Additional Safeguards for Vulnerable Subjects  
The present research will involve subjects who might be vulnerable to coercion or undue 
influence. As required in 45CFR46.111 (b), we recommend that sites utilize additional 
safeguards to protect the rights and welfare of these subjects. Such safeguards might include 
but are not limited to: a) assessment of the potential subject’s capacity to provide informed 
consent, b) the availability of the LAR to monitor the subject’s subsequent participation and 
withdrawal from the study; c) augmented consent processes. The specific nature of the 
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 10.9     Confidentiality  
Federal regulations at 45 CFR 46 111 (a) (7) requires that when appropriate, there are 
adequate provisions to protect the privacy of subjects and to maintain the confidentiality of data. 
To maintain confidentiality, all laboratory specimens, evaluation forms, and reports will be 
identified only by a coded number.  The coded number will be generated by a computer, and 
only the study team will have access to the codes.  All records will be kept in a locked, 
password protected computer.  All computer entry and networking programs will be done with 
coded numbers only.  All paper case report forms will be maintained inside a locked office.  
Study information will not be released without the written permission of the patient, except as 
necessary for monitoring by the National Heart, Lung, and Blood Institute, and the PETAL 
Clinical Coordinating Center.  
11. Adverse Events  
11.1 Safety Monitoring  
Assuring patient safety is an essential component of this protocol. Each participating 
investigator has primary responsibility for the safety of the individual participants under his 
or her care. The Investigators will determine daily if any adverse events occur during the 
period from enrollment through study day 10 (five days after completion of the study drug 
infusions) or hospital discharge, whichever occurs first and will determine if such adverse 
events are reportable.   
The following adverse events will be considered reportable and thus collected in the adverse 
event case report forms:  
  
▪ 
Serious adverse events  
▪ 
Non-serious adverse events that are considered by the investigator to be related to 
study procedures or of uncertain relationship (Appendix E)  
▪ 
Study-specific clinical outcomes (Primary and Secondary Outcomes and 
Assessments During the Study), including serious adverse events such as organ 
failures and death, are systematically recorded in the case report forms and are 
exempt from adverse event reporting unless the investigator deems the event to 
be related to the administration of study drug or the conduct of study procedures 
(or of uncertain relationship) as outlined in Appendix E.   
After randomization, adverse events must be evaluated by the investigator. If the adverse event 
is judged to be reportable, as outlined above, then the investigator will report to the CCC their 
assessment of the potential relatedness of each adverse event to the study drug or protocol 
procedure via electronic data entry. Investigators will assess if there is a reasonable possibility 
that the study procedure caused the event, based on the criteria outlined in Appendix E. 
Investigators will also consider if the event is unexpected.  Unexpected adverse events are 
events not listed in the protocol and the investigator brochure for intravenous Vitamin C or 
Acetaminophen. Investigators will also determine if adverse events are unanticipated given the 
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If a patient's treatment is discontinued as a result of an adverse event, study site personnel 
must report the circumstances and data leading to discontinuation of treatment in the adverse 
event case report forms.    
11.2 Serious Adverse Events  
Serious adverse event collection begins after randomization and study procedures have been 
initiated.  If a patient experiences a serious adverse event after consent, but prior to 
randomization or starting study procedures, the event will NOT be collected. Study site 
personnel must alert the CCC of any serious and study procedure related adverse event 
within 24 hours of investigator awareness of the event.  Alerts issued via telephone are to be 
immediately followed with official notification on the adverse event case report form. See 
Appendix E for reporting timelines for serious, unexpected, study related events (SAEs) and 
serious, unexpected suspected adverse reactions (SUSARs).  
As per the FDA and NIH definitions, a serious adverse event is any adverse event that results in 
one of the following outcomes:  
▪ Death   
▪ A life-threatening experience (that is, immediate risk of dying)  
▪ Prolonged inpatient hospitalization or re-hospitalization  
As per http://www.fda.gov/Safety/MedWatch/HowToReport/ucm053087.htm: Report if 
admission to the hospital or prolongation of hospitalization was a result of the adverse 
event. Emergency room visits that do not result in admission to the hospital should be 
evaluated for one of the other serious outcomes (e.g., life-threatening; required 
intervention to prevent permanent impairment or damage; other serious medically 
important event).  
▪ Persistent or significant disability/incapacity  
As per http://www.fda.gov/Safety/MedWatch/HowToReport/ucm053087.htm: Report if the 
adverse event resulted in a substantial disruption of a person's ability to conduct normal 
life functions, i.e., the adverse event resulted in a significant, persistent or permanent 
change, impairment, damage or disruption in the patient's body function/structure, physical 
activities and/or quality of life.  
Reportable serious adverse events that may not result in death, be life-threatening, or require 
hospitalization may be considered serious adverse events when, based upon appropriate 
medical judgment, they may jeopardize the patient and may require medical or surgical 
intervention to prevent one of the outcomes listed in this definition.  
Serious adverse events will be collected during the first 10 study days or until hospital  
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Appendices  
A. Schedule of Events   
 
  
  


  
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B. Sequential (Sepsis-related) Organ Failure Assessment (SOFA) Score   
SOFA Score  
0  
1  
2  
3  
4  
Pulmonary (PaO2/FiO2, 
mmHg)  
≥ 400  
< 400  
< 300  
< 200  
< 100  
Coagulation (Platelets x 
103/mm3)  
≥ 150  
< 150  
< 100  
< 50  
< 20  
Liver (Total Bilirubin, 
mg/dL)  
< 1.2  
1.2 – 1.9  
2.0 – 5.9  
6.0 – 11.9  
≥ 12.0  
Cardiovascular (Mean 
Arterial Pressure, 
mmHg)  
≥ 70  
< 70  
Dopamine ≤ 5  
or dobutamine   
(any dose)*  
Dopamine > 5 or 
epinephrine ≤ 0.1 
or norepinephrine 
≤ 0.1*  
Dopamine > 15 or 
epinephrine > 0.1 or 
norepinephrine > 
0.1*  
Renal (Serum 
Creatinine, mg/dL)  
< 1.2  
1.2 – 1.9  
2.0 – 3.4  
3.5 – 4.9  
≥ 5.0  
*Adrenergic agents administered for at least 1 hour, doses given as μg/kg/min  
  


  
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C. Ventilator Management for Patients with ARDS  
PART I: VENTILATOR SETUP AND ADJUSTMENT  
1. Calculate predicted body weight (PBW) in kilograms  
Males = 50 + 2.3 [height (inches) - 60]  
Females = 45.5 + 2.3 [height (inches) -60]  
2. Controlled ventilation mode required during NMB.  After NMB, select any ventilator mode 
capable of delivering the prescribed VT (6ml/kg PBW, +/- 2 ml/kg)   
  
 
OXYGENATION GOAL: PaO2 55-80 mmHg or SpO2 88-95%  
1. Use a minimum PEEP of 5 cm H2O.   
2. Adjust FiO2 or PEEP upward within 5 minutes of consistent measurements below the 
oxygenation target range  
3. Adjust FiO2 or PEEP downward within 30 minutes of consistent measurements above the 
oxygenation target range.    
4. The below PEEP strategy FiO2/PEEP table should be used in all patients.  
 
PEEP/FiO2  
FiO2  
.30  .40  .40  
.50  
.60  .70  
.80  .90  
1.0  
PEEP  
5  
5  
 8  
8-10  10  10-14  14  14-18  
18-24  
 
PLATEAU PRESSURE GOAL: ≤ 30 cm H2O  
Check Pplat (0.5 second inspiratory pause), at least q 4h and after each change in PEEP or VT.  
• 
If Pplat > 30 cm H2O: decrease VT by 1ml/kg steps (minimum = 4 ml/kg).  
• 
If Pplat < 25 cm H2O and VT< 6 ml/kg, increase VT by 1 ml/kg until Pplat > 25 cm H2O 
or VT = 6 ml/kg.  
• 
If “severe dyspnea” (more than 3 double breaths per minute or airway pressure 
remains at or below PEEP level during inspiration), then raise VT to 7 or 8 ml/kg PBW if 
Pplat remains below 30.  If Pplat exceeds 30 cm H20, then revert to lower VT and 
consider more sedation.  
pH GOAL: ≥ 7.30  
• 
If pH < 7.30: May give NaHCO3 (neither encouraged nor discouraged)  
• 
If pH < 7.15: VT may be raised and Pplat limit suspended (neither encouraged nor 
discouraged)  
• 
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I:E RATIO GOAL: Recommend that duration of inspiration be  duration of expiration.  
PART II: WEANING  
Conduct a SPONTANEOUS BREATHING TRIAL daily when:  
• 
FiO2 ≤ 0.40 and PEEP ≤ 8 
• 
PEEP and FiO2 ≤ values of previous day 
• 
Systolic BP ≥ 90 mmHg and without vasopressor support 
• 
Spontaneous respirations  
SPONTANEOUS BREATHING TRIAL (SBT):  
If all above criteria are met initiate a trial of UP TO 120 minutes of spontaneous breathing with 
FiO2 < 0.5 using any of the following approaches:  
• 
Pressure support < 5cm H2O, PEEP < 5cm H2O  
• 
CPAP < 5 cm H2O   
• 
T-piece  
• 
Tracheostomy mask   
Assess for tolerance using the following:  
• 
SpO2 ≥ 90: and/or PaO2 ≥ 60 mmHg  
• 
Spontaneous VT ≥ 4 ml/kg PBW  
• 
RR ≤ 35/min  
• 
pH ≥ 7.3  
• 
No respiratory distress (distress = 2 or more of the following):  
o 
HR > 120% of baseline 
o 
Marked accessory muscle use  
o 
Abdominal paradox  
o 
Diaphoresis 
o 
Marked dyspnea  
If tolerated for at least 30 minutes, consider extubation.  
If not tolerated resume pre-weaning settings.  
  
  
 
 


  
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D. On-Study Fluid Management for Patients with ARDS  
This protocol should be initiated after the initial resuscitation.  
o Discontinue maintenance fluids.  
o Continue medications and nutrition.  
o Manage electrolytes and blood products per usual practice.  
o For shock, use any combination of fluid boluses# and vasopressor(s) to achieve MAP ≥ 
60 mmHg as fast as possible.  Wean vasopressors as quickly as tolerated beginning four 
hours after blood pressure has stabilized.  
o Withhold diuretic therapy in renal failure § and until 12 hours after last fluid bolus or 
vasopressor given.  
  
  
CVP  
(recommend)  
  
  
  
PAOP  
(optional)  
  
MAP > 60 mm Hg AND off vasopressors for > 12 hours  
Average urine output < 0.5 
ml/kg/hr  
Average urine output > 0.5 
ml/kg/hr  
>8  
> 12  
Furosemide* 
Reassess in 1 hour  
Furosemide* 
Reassess in 4 hours  
4-8  
8-12  
Give fluid bolus as fast as 
possible# Reassess in 1 hour  
  
< 4  
< 8  
No intervention 
Reassess in 4 hours  
  
  
§ Renal failure is defined as dialysis dependence, oliguria with serum creatinine > 3mg/dl, or oliguria with serum 
creatinine 0-3 with urinary indices indicative of acute renal failure as in the original FACTT trial.  
# Recommended fluid bolus= 15 mL / kg crystalloid (round to nearest 250 mL) or 1 Unit packed red cells or 25 
grams albumin  
*Recommended Furosemide dosing = begin with 20 mg bolus or 3 mg / hr infusion or last known effective dose.  
Double each subsequent dose until goal achieved (oliguria reversal or intravascular pressure target) or maximum 
infusion rate of 24 mg / hr or 160 mg bolus reached.  Do not exceed 620 mg / day.  Also, if patient has heart failure, 
consider treatment with dobutamine.  
  
NOTE: For patients without a central venous catheter, no fluid gain over the first 7 study days is 
recommended once patients’ blood pressure has stabilized.  Stable blood pressure is defined as 
no requirement for either vasopressors or a fluid bolus to support blood pressure for 12 or more 
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E. Adverse Event Reporting and Unanticipated Events  
As noted in section 11, investigators will report all adverse events that are serious and study 
drug or study procedure related (or of uncertain relatedness) to the CCC within 24 hours. The 
CCC will then notify the NHLBI and Central Institutional Review Board (cIRB).   
The Medical Monitor at the CCC will work collaboratively with the reporting investigator to 
determine if a serious adverse event has a reasonable possibility of having been caused by the 
study drug or study procedure, as outlined in 21 CFR 312.32(a)(1), and below. The Medical 
Monitor will be unblinded and will also determine if the event is unexpected for Acetaminophen or 
Vitamin C, depending on treatment assignment. An adverse event is considered “unexpected” if 
it is not listed in the investigator brochure or the study protocol (21 CFR 312.32(a)). If a 
determination is made that a serious adverse event has a reasonable possibility of having been 
caused by a study procedure or the study drug, it will be classified as a suspected adverse 
reaction. If the suspected adverse reaction is unexpected, it will be classified as a serious 
unexpected suspected adverse reaction (SUSAR).   
The CCC will report all unexpected deaths, serious and treatment related adverse events, and 
SUSARs to the DSMB, NHLBI, and cIRB within 7 days after receipt of the report from a clinical 
site. A written report will be sent to the NHLBI, DSMB and the cIRB within 15 calendar days. The 
DSMB will also review all adverse events and clinical outcomes during scheduled interim 
analyses. The CCC will distribute the written summary of the DSMB’s periodic review of adverse 
events to the cIRB in accordance with NIH guidelines 
(http://grants.nih.gov/grants/guide/noticefiles/not99-107.html).   
E.1. Unanticipated Problems (UP)  
Investigators must also report Unanticipated Problems, regardless of severity, associated with 
study procedures within 24 hours. An unanticipated problem is defined as follows: any incident, 
experience, or outcome that meets all of the following criteria:   
• 
Unexpected, in terms of nature, severity, or frequency, given the research procedures that 
are described in the protocol-related documents, such as the IRB-approved research 
protocol and informed consent document; and the characteristics of the subject population 
being studied;   
• 
Related or possibly related to participation in the research. Possibly related means there is a 
reasonable possibility that the incident, experience, or outcome may have been caused by 
the procedures involved in the research;   
• 
Suggests that the research places subjects or others at a greater risk of harm (including 
physical, psychological, economic, or social harm) than was previously known or 
recognized.   
E.2. Determining Relationship of Adverse Events to Procedures  
Investigators will be asked to grade the strength of the relationship of an adverse event to study 
procedures as follows:   
• 
Definitely Related: The event follows: a) A reasonable, temporal sequence from a study 
procedure; and b) Cannot be explained by the known characteristics of the patient’s 


  
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clinical state or other therapies; and c) Evaluation of the patient’s clinical state indicates 
to the investigator that the experience is definitely related to study procedures.   
• 
Probably or Possibly Related: The event should be assessed following the same criteria 
for “Definitely Related”. If in the investigator’s opinion at least one or more of the criteria 
are not present, then “probably” or “possibly” associated should be selected.   
• 
Probably Not Related: The event occurred while the patient was on the study but can 
reasonably be explained by the known characteristics of the patient’s clinical state or 
other therapies.   
• 
Definitely Not Related: The event is definitely produced by the patient’s clinical state or by 
other modes of therapy administered to the patient.   
• 
Uncertain Relationship: The event does not meet any of the criteria previously outlined.   
E.3. Clinical Outcomes that may be Exempt from Adverse Event Reporting  
Study-specific clinical outcomes of sepsis, as outlined in Section 8.1 and 8.3 (Primary and 
Secondary Outcomes) and Section 7.2 (Assessments During the Study) are exempt from 
adverse event reporting unless the investigator deems the event to be related to the study 
procedures (or of uncertain relationship) or if the event leads to discontinuation of study 
procedures. The following are examples of events that will be considered study specific clinical 
outcomes:   
• 
Death not related to the study procedures   
• 
Cardiovascular events: need for vasoactive drugs or fluids for hypotension or 
hypotension not temporally related to study drug infusion  
• 
Respiratory events: decreased PaO2/FiO2, hypoxia, worsening acute respiratory 
distress syndrome, or respiratory failure.   
• 
Hepatic events: hepatic injury or liver dysfunction that leads to an increase from baseline 
in the serum level of bilirubin.   
• 
Renal events: renal failure, renal insufficiency, or renal injury that leads to an increase 
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F. Alcohol Use Screen Audit C  
1. How often do you have a drink containing alcohol?  
 Never (0 points)  
 Monthly or less (1 point)  
 2-4 times a month (2 points)  
 2-3 times a week (3 points)  
 4 or more times a week (4 points)  
2. How many standard drinks containing alcohol do you have on a typical day?  
  1 or 2 (0 points)  
  3 or 4 (1 point)  
  5 or 6 (2 points)  
  7 or 9 (3 points)  
  10 or more (4 points)  
3. How often do you have six or more drinks on one occasion?  
  Never (0 points)  
  Less than monthly (1 point)  
  Monthly (2 points)  
  Weekly (3 points)  
  Daily or almost daily (4 points)  
TOTAL POINTS:  ______  
If total > 4, patient is excluded from the study.  
   
  
  


  
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G. Fever Management Guidelines   
Use of nonsteroidal anti-inflammatory drugs (NSAID)s or Cyclooxygenase-2 inhibitors to treat 
fever will be strongly discouraged during the 5 day study drug administration and the use of 
these medications for other indications will also be discouraged. No restriction will be placed on 
the use of aspirin in doses of less than or equal to 325 mg per day. The use of open-label 
acetaminophen will not be permitted while receiving study medication. Once the course of study 
drug administration has been completed, open-label acetaminophen can be administered at the 
discretion of the treating clinician. Physical cooling will be used at the discretion of the clinical 
team per local practice preferences (modified from Reference 38).   
   
  
 


  
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Fowler, A.A., 3rd, et al., Effect of Vitamin C Infusion on Organ Failure and  
Biomarkers of Inflammation and Vascular Injury in Patients With Sepsis and Severe 
Acute Respiratory Failure: The CITRIS-ALI Randomized Clinical Trial. JAMA, 2019. 
322(13): p. 1261-1270.  
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Masse MH, Ménard J, Sprague S, et al. Lessening Organ dysfunction with VITamin C 
(LOVIT): protocol for a randomized controlled trial. Trials. 2020;21(1):42. 


  
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June 8, 2021 
 
 
 
 
 
 
 
 
 
 
Pg.  61     
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Boutaud, O., et al., Acetaminophen inhibits hemoprotein-catalyzed lipid peroxidation and 
attenuates rhabdomyolysis-induced renal failure. Proc Natl Acad Sci U S A, 2010. 
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36. 
Ouellet, M. and M.D. Percival, Mechanism of acetaminophen inhibition of 
cyclooxygenase isoforms. Arch Biochem Biophys, 2001. 387(2): p. 273-80.  
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Anderson, B.J., Paracetamol (Acetaminophen): mechanisms of action. Paediatr Anaesth, 
2008. 18(10): p. 915-21.  
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Janz, D.R., et al., Randomized, placebo-controlled trial of acetaminophen for the 
reduction of oxidative injury in severe sepsis: the Acetaminophen for the Reduction of 
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Watkins, P.B., et al., Aminotransferase elevations in healthy adults receiving 4 grams of 
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ICF_004.pdf: 
PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen  in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  07/07/2022 
  
 
1 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
Part 1 of 2: MASTER CONSENT 
 
Name of participant: __________________________________________________________ Age: ___________ 
 
You are being invited to take part in a research study. This form will tell you about this study. Please read this 
form carefully and ask any questions you may have. This study is a multi-site study, meaning it will take place 
at several different places. Because this is a multi-site study this consent form includes two parts. Part 1 of this 
consent form is the Master Consent and includes information that applies to all study sites. Part 2 of the 
consent form is the Study Site Information and includes information specific to the study site where you are 
being asked to participate. Both parts together are the legal consent form and must be provided to you. 
 
Key information about this study: 
You are being asked to take part in this study because you have a severe infection (sepsis). In this study we are 
trying to understand whether acetaminophen (Tylenol)  helps patients recover from sepsis compared to patients 
treated with a placebo (sugar water).  Sepsis causes damage to red blood cells which can release a protein into 
the blood called hemoglobin.  Hemoglobin can cause injury to the blood vessels and organs of patients with 
sepsis. In laboratory studies and in small studies of patients with sepsis, acetaminophen  can stop some of the 
effects of hemoglobin. This may then limit injury to blood vessels and organs, and help people recover from 
sepsis.  
Acetaminophen has been used many times in humans with good safety. Possible risks of acetaminophen include 
liver injury or lower blood pressure. You will be carefully monitored for these if you choose to participate.  
If you agree to be in this study you could receive Tylenol or a placebo through an IV for up to 5 days while you 
are in the ICU. We will collect information from your medical record while you are in the hospital.  
Acetaminophen  is not given to all sepsis patients outside of research, and if you do not join, your doctor will 
choose all therapies. 
You may choose not to be in this study. If you choose not to be in the study, you will receive the medical 
treatment that you are receiving now and anything else that your doctors think might be helpful.  You can stop 
being in this study at any time.  
If we learn something while doing this study that could change the risks or benefits of you being in it, we will tell 
you so that you can decide if you want to stay in or join this study.  
The word “you” in this form refers to the person who will be in the study. Your legal representative will be asked 
to read and sign this consent form to give permission for you to participate if you are unable to do so yourself.    
Key Information:  
The first section of this document contains some key points that the research team thought you 
would find important. The study is described in more detail after this section. If you do not 
understand something, please ask someone.  
Date of IRB Approval: 07/27/2022
Date of Expiration: 07/05/2023


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen  in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  07/07/2022 
  
 
2 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
 
 
What is the purpose of this study?  
We are studying if acetaminophen helps people with sepsis. The study is paid for by the National Institutes of 
Health.  
Sepsis can lower your blood pressure and worsen the function of your internal organs, including the lungs and 
kidneys. These effects of sepsis can be reversible (can be fixed). We are studying whether acetaminophen 
improves your blood pressure and/or the function of your lungs and kidneys.  
If you agree to join the study, you will be assigned by chance (like a coin toss) to one of two groups.. One group 
gets acetaminophen intravenously (through a small tube in a vein) for 5 days. The  other group gets a placebo 
(simple sugar water that does not contain acetaminophen) for 5 days. All groups will receive standard treatment 
for sepsis, including antibiotics, as directed by your treating physicians.  
What will happen and how long will you be in the study? 
 Overview: We seek 450 people with sepsis to join this study in about 50 hospitals. Your hospital is one of 
these hospitals and expects to enroll 15  or more people.  If you choose to participate in this study, you will 
be in the study for three months (90 days).   
 Review of your medical information: First, we reviewed your medical information to ensure you are eligible 
to join the research study.  
 Pregnancy Check: If you agree to join this study and  are a woman who can have children, we will do a 
pregnancy test before you join the study, if your doctor has not already checked this as part of your normal 
care. If you are pregnant, you may not join the study. 
 Laboratory Tests: We will check your liver function levels in your blood before you join the study, if your 
doctor has not already checked this as part of your normal care. If you already have very high liver tests you 
will not be allowed to join the study.  
 
 
Detailed Information: 
The rest of this document includes detailed information about this study (in addition to the 
information listed above). 
Date of IRB Approval: 07/27/2022
Date of Expiration: 07/05/2023


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen  in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  07/07/2022 
  
 
3 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
If you are eligible to be in the study, and you agree to participate:  
 
 
Side effects and risks that you can expect if you take part in this study:  
Acetaminophen: At high doses, acetaminophen can injure the liver, especially in patients who already have 
liver problems. For your safety, we will monitor your liver tests and will stop the study medicine if your liver 
tests suggest that your liver function is worsening. Stopping acetaminophen usually results in liver tests 
returning to normal without permanent damage.  
Less commonly, acetaminophen can lower blood pressure. We will watch your blood pressure after each dose 
of the study medicine. If your blood pressure goes down too much, your doctors may treat the low blood 
pressure as part of regular clinical care.  
An allergic reaction is possible but rare; most are mild, with very few allergic reactions being life threatening. If 
you develop an allergic reaction to acetaminophen, your doctors will treat it.   
Drawing Blood: There are no major risks associated with drawing blood. We will usually take blood from tubes 
that are already in your vein or artery. If you do not have a tube in a vein or artery, then we will get the blood 
with a needle. You may experience minor discomfort, bruising or soreness from the needle. Drawing blood 
through a needle very rarely causes infection. Blood draws will be done by a trained professional.  
Health Information Privacy: To ensure your privacy, your health information will be handled securely by the 
participating hospitals, the Clinical Data Center at Massachusetts General Hospital and the National Institutes of 
Health. 
Risks that are not known: 
We do not know if your blood pressure will be higher or lower or if the function of your lungs and kidneys will be 
better or worse if you choose to be in this study. We also do not know whether your risk of dying will be higher 
or lower if you choose to be in this study.  If you are in the group that receives acetaminophen, there may be 
unknown side effects. If you are in the group that receives placebo, you will not get the possible benefit of 
acetaminophen that this study is testing. If we learn something new that affects the risks or benefits of being in 
You will receive either 
Acetaminophen or a 
Placebo. 
   
This is assigned by chance 
(like a coin toss). You, your 
doctors, and the research 
team will not know which 
one you will get.  
ACETAMINOPHEN 
225 patients will receive acetaminophen through a tube in the 
vein every 6 hours for up to 5 days while in the intensive care 
unit  
PLACEBO  
 225 patients will receive placebo through a tube in the vein every 6 
hours for up to 5 days while in the intensive care unit. A placebo is an 
inactive liquid that looks the active drug but does not contain any 
medicine. 
Date of IRB Approval: 07/27/2022
Date of Expiration: 07/05/2023


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen  in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  07/07/2022 
  
 
4 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
this study, we will change the study plan as needed. We will also make sure you are told about this new 
information while you are in the study. 
Also, because acetaminophen is being used in this trial, fever cannot be treated with this medication. If you 
develop a fever, other medicines and cooling methods will be used if your doctors feel your fever should be 
treated, which may be different from standard practice. 
Good effects that might result from this study  
Early evidence suggests that acetaminophen may improve recovery from sepsis, although we are not sure of 
this. We cannot know whether acetaminophen will make your recovery better, worse, or the same. Your 
participation in this study helps us learn more about treating patients like you with sepsis and may help other 
patients in the future. The tests we do on your blood will not help you directly.  
During the study: 
 We will talk with your doctors. The research team will tell your doctors about the study, and your doctors 
will decide on all of your additional treatments based on your needs.  
 We will review your medical record. We will look at your medical records or check on you to see how you 
are doing.   We will collect information like your blood pressure, medical history, and test results. We may 
ask questions to you or your doctors if something is not clear. If you go to another health care facility, we 
may contact you or the health care facility to find out how you are doing. 
 We will collect some of your blood. We will 
collect up to 2 tablespoons of blood at the 
start of the study. We will also collect up to 1 
tablespoon of blood after 2 days, and then up 
to 2 tablespoons of blood after 3 days in the 
study. Some of the blood samples will be 
stored for future studies of serious illness and 
other conditions. Samples will not have your 
name on them, and instead use a coded study number to protect your privacy. We will not include your 
name or other identifying information with the samples ever. Only your hospital study team and the 
research study Coordinating Center at Massachusetts General Hospital will know your coded study number. 
These numbers will be kept private.  
 We will collect some of your urine. We will collect some of your urine (pee) three times during the study: at 
the start of the study, and on days 2 and 3. These samples will be labeled with a coded study number and 
kept private; the same as the blood samples.   
 We will monitor your liver function tests. We will measure levels of two liver function tests at the start of 
the study, and then after 2, 3, 4, 5, and 7 days in the study. We do not anticipate that this will require an 
extra blood draw because this testing can usually be done in the clinical laboratory at your hospital on blood 
samples that your regular doctors are already sending. Rarely, this may require an extra blood draw. We are 
doing this because acetaminophen can affect the function of your liver. 
STUDY BLOOD COLLECTION PLAN: 
Date of IRB Approval: 07/27/2022
Date of Expiration: 07/05/2023


PETAL Network Central Institutional Review Board 
Informed Consent Document for Research 
MASTER CONSENT 
Study Title: 
Acetaminophen  in Sepsis: Targeted Therapy to Enhance Recovery (ASTER) 
Version Date:  07/07/2022 
  
 
5 
This box is for 
CIRB USE ONLY 
VERSION 6 
Do not edit or delete 
 We will check to see how you are doing in one month and in three months. An important part of this study 
is to find out how you are doing after you leave the hospital. To make sure we can reach you, we will record 
your information, such as contact phone numbers and social security number. This information will be kept 
private. 
o If you are still in the hospital, we will check on you in person or by telephone. 
o If you are out of the hospital, we will contact you using the contact information you provide. 
o If we cannot reach you, we will check hospital and national health records using the information you 
provide. 
Other treatments you could get if you decide not to be in this study 
Taking part in this study is voluntary. You will receive the care you need whether or not you join the study or if 
you withdraw at any time. Acetaminophen is not given to all sepsis patients outside of research, and if you do 
not join, your doctor will choose all therapies.  
Reasons why the study doctor may take you out of this study 
The study doctor may take you out of this study if his/her judgment suggests that is better for you. This decision 
may be based on new information about your condition or the study risks and benefits. 
What will happen if you decide to stop being in this study? 
You can stop being in this study at any time. If you decide to stop, tell your study doctor or a member of the 
study team. Deciding to not be part of the study is completely up to you. This will not affect the care you 
receive. 
If you stop being in the study, we will store your blood samples unless you request they be destroyed. We will 
continue to collect information from your medical record unless you ask us not to.  
Clinical Trials Registry  
A description of this clinical trial is available on www.clinicaltrials.gov, which will not include information that 
can identify you. Eventually, the Web site will include a summary of the study results but no personal 
information. You can search this Web site at any time. 
Privacy: 
Any samples and information about you may be made available to others to use for research.  To protect your 
privacy, we will not release your name.  You will not receive any benefit as a result of the tests done on your 
samples.  These tests may help us or other researchers learn more about the causes, risks, treatments, or how to 
prevent this and other health problems. 
Study Results:  
We will share the study results with you by letter within 3 months of the publication of the manuscripts 
reporting the results of the trial.   
 
Date of IRB Approval: 07/27/2022
Date of Expiration: 07/05/2023