Supplementary Appendix
This appendix has been provided by the authors to give readers additional information about their work.
Supplement to: Heald AE, Charleston JS, Iversen PL, et al. AVI-7288 for Marburg virus in nonhuman primates
and humans. N Engl J Med 2015;373:339-48. DOI: 10.1056/NEJMoa1410345
TABLE OF CONTENTS
Timing of Plasma Sampling in AVI-7288 Pharmacokinetics and Pharmacodynamics Study in
Infected Macaques ..................................................................................................................3
Timing of Plasma Sampling in AVI-7288 Pharmacokinetics Study in Uninfected Macaques .......4
Bioanalytic Assay for Determining AVI-7288 Levels in Cynomolgus Monkey Plasma ................5
Bioanalytic Assay for Determining AVI-7288 Levels in Human Plasma .......................................6
Figure S1. Area under the Curve and Maximum Plasma Concentration in Infected and
Uninfected Nonhuman Primates .............................................................................................7
Figure S2. Maximum Plasma Concentration in Uninfected Humans ..............................................8
Figure S3. Clearance in Infected and Uninfected Nonhuman Primates ..........................................9
Figure S4. Clearance in Uninfected Humans .................................................................................10
Figure S5. Area under the Curve and Clearance of AVI-7288 in Infected and Uninfected
Nonhuman Primates ..............................................................................................................11
Figure S6. Maximum Plasma Concentration after Intramuscular and Intravenous Administration
of AVI-7288 in Uninfected Nonhuman Primates .................................................................12
Table S1. Summary of Studies in Nonhuman Primates .................................................................13
Table S2: Characteristics of Healthy Volunteers in Multiple Ascending Dose Study ..................15
Table S3. Summary of Adverse Events Occurring during Study Period by Severity ...................16
Table S4. Summary of Treatment-Related Adverse Events Occurring during Study Period by
Severity .................................................................................................................................21
Table S5. Pharmacokinetic Parameters of AVI-7288 in Infected Nonhuman Primates ................23
Table S6. Pharmacokinetic Parameters of AVI-7288 in Uninfected Nonhuman Primates ...........24
Table S7. Pharmacokinetic Parameters of AVI-7288 in Uninfected Humans...............................25
Table S8. Median, Geometric Mean, and 5th and 95th Percentile Values from 10,000 Simulation
AUC0-inf Values in Infected Nonhuman Primates and Uninfected Humans .........................26
2
Timing of Plasma Sampling in AVI-7288 Pharmacokinetics and Pharmacodynamics Study
in Infected Macaques
Plasma was collected for pharmacokinetic sampling at the following time points in the AVI-7288
groups:
•
•
•
•
•
•
•
Day 1 (infection day): Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 1h ± 5 min,
2h ± 7 min, 4h ± 10 min, 8h ± 15 min, and 24 ± 1h after AVI-7288 administration
Day 5: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 1h ± 5 min, 2h ± 7 min,
4h ± 10 min, 8h ± 15 min, and 24 ± 1h after AVI-7288 administration
Day 10: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 1h ± 5 min, 2h ± 7 min,
4h ± 10 min, 8h ± 15 min, and 24 ± 1h after AVI-7288 administration
Day 14: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 1h ± 5 min, 2h ± 7 min,
4h ± 10 min, 8h ± 15 min, and 24 ± 1h after AVI-7288 administration
Day 16: 48 ± 2h after AVI-7288 administration on day 14
Day 17: 72 ± 2h after AVI-7288 administration on day 14
Day 18: 96 ± 2h after AVI-7288 administration on day 14
3
Timing of Plasma Sampling in AVI-7288 Pharmacokinetics Study in Uninfected Macaques
Plasma was collected for pharmacokinetic sampling at the following time points in the singledose AVI-7288 3.75 mg/kg group:
•
Day 1: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 45 ± 4 min, 1h ± 5 min,
2h ± 7 min, 4h ± 10 min, 8h ± 15min, 10h ± 20 min, 12h ± 20 min, and 24 ± 1h after
AVI-7288 administration
Plasma was collected for pharmacokinetic sampling at the following time points in the groups
that received AVI-7288 7.5 mg/kg or AVI-7288 15 mg/kg daily for 14 days:
•
•
•
•
•
•
•
Day 1 (first dosing day): Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 45 ± 4 min,
1h ± 5 min, 2h ± 7 min, 4h ± 10 min, 8h ± 15min, 10h ± 20 min, 12h ± 20 min, and
24 ± 1h after AVI-7288 administration
Day 5: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 45 ± 4 min,
1h ± 5 min, 2h ± 7 min, 4h ± 10 min, 8h ± 15min, 10h ± 20 min, 12h ± 20 min, and
24 ± 1h after AVI-7288 administration
Day 10: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 45 ± 4 min,
1h ± 5 min, 2h ± 7 min, 4h ± 10 min, 8h ± 15min, 10h ± 20 min, 12h ± 20 min, and
24 ± 1h after AVI-7288 administration
Day 14: Immediately pre-dose; 15 ± 2 min, 30 ± 3 min, 45 ± 4 min,
1h ± 5 min, 2h ± 7 min, 4h ± 10 min, 8h ± 15min, 10h ± 20 min, 12h ± 20 min, and
24 ± 1h after AVI-7288 administration
Day 16: 48 ± 1h after AVI-7288 administration on day 14
Day 17: 72 ± 1h after AVI-7288 administration on day 14
Day 18: 96 ± 1h after AVI-7288 administration on day 14
4
Bioanalytic Assay for Determining AVI-7288 Levels in Cynomolgus Monkey Plasma
The method for determination of AVI-7288 in monkey plasma employed a detector reagent
comprised of a complementary oligonucleotide probe with terminal fluorescein label, which
directly hybridized to the AVI-7288 analyte in monkey plasma after TRIzol LS extraction and
alcohol precipitation. After hybridization, the resulting DNA-morpholino molecule (hybrid) was
subjected to analysis by capillary gel electrophoresis with laser-induced fluorescence detection.
Quantification of the fluorescent hybrid by capillary gel electrophoresis hybridization was
determined by normalizing the fluorescence intensity of each hybrid peak to an unrelated
fluorescent internal standard peak, whereby changes in the normalized value were proportional
to changes in AVI-7288 analytic concentrations in different samples. The hybrid peak
fluorescence correlated to analyte quantity in a range of 1.9 ng/mL to 29.5 ng/mL plasma.
Concentrations below the lower limit of quantitation were reported as BLOQ.
The method was validated by meeting acceptance criteria for selectivity, range of response,
dilution linearity, lower and upper limits of quantitation, intra- and inter-assay accuracy and
precision, multiple stability assessments, and recovery.
5
Bioanalytic Assay for Determining AVI-7288 Levels in Human Plasma
The method for determination of AVI-7288 in human plasma employed a detector reagent
comprised of a complementary oligonucleotide probe with terminal fluorescein label, which
directly hybridized to the AVI-7288 analyte in human plasma after TRIzol LS extraction and
alcohol precipitation. After hybridization, the resulting DNA-morpholino molecule (hybrid) was
subjected to analysis by capillary gel electrophoresis with laser-induced fluorescence detection.
Quantification of the fluorescent hybrid by capillary gel electrophoresis hybridization was
determined by normalizing the fluorescence intensity of each hybrid peak to an unrelated
fluorescent internal standard peak, whereby changes in the normalized value were proportional
to changes in AVI-7288 analytic concentrations in different samples. The hybrid peak
fluorescence correlated to analyte quantity in the range of 2.9 to 47.1 ng/mL in plasma.
Concentrations below the lower limit of quantitation were reported as BLOQ.
The method was validated by meeting acceptance criteria for selectivity, range of response,
dilution linearity, lower and upper limits of quantitation, intra- and inter-assay accuracy and
precision, multiple stability assessments, and recovery.
6
Figure S1. Area under the Curve and Maximum Plasma Concentration in Infected and
Uninfected Nonhuman Primates
(A) Area under the plasma concentration-time curve from time 0 to 24 hours after the start of
infusion of study drug (AUC0-24) and (B) maximum plasma concentration (Cmax) were generally
dose-proportional over the range of doses tested (3.75, 7.5, and 15 mg/kg) on day 1 in infected
nonhuman primates (red) and uninfected nonhuman primates (blue) in the dedicated nonhuman
primate pharmacokinetic studies. Values depicted are mean ± standard deviation.
7
Figure S2. Maximum Plasma Concentration in Uninfected Humans
Maximum plasma concentration (Cmax) was generally dose-proportional over the range of doses
tested (1, 4, 8, 12, and 16 mg/kg) in uninfected humans in the multiple ascending dose study at
day 1 (red) and day 14 (blue). Values depicted are mean ± standard deviation.
8
Figure S3. Clearance in Infected and Uninfected Nonhuman Primates
Plasma clearance was independent of dose over the range of doses tested (3.75, 7.5, and 15
mg/kg) in infected nonhuman primates (red) and uninfected nonhuman primates (blue) on day 1
in the dedicated nonhuman primate pharmacokinetic studies. Slope was not significantly
different from 0 for infected or uninfected nonhuman primates (p=0.307 and p=0.222,
respectively).
9
Figure S4. Clearance in Uninfected Humans
Plasma clearance was independent of dose over the range of doses tested (1, 4, 8, 12, and 16
mg/kg) in uninfected humans on day 14 in the multiple ascending dose study. Slope was not
significantly different from 0 (p=0.06).
10
Figure S5. Area under the Curve and Clearance of AVI-7288 in Infected and Uninfected
Nonhuman Primates
(A) Area under the plasma concentration-time curve from time 0 to 24 hours after the start of
infusion of study drug (AUC0-24) and (B) plasma clearance of AVI-7288 15 mg/kg were similar
in infected nonhuman primates (red) and uninfected nonhuman primates (blue) at day 1, day 5,
day 10, and day 14, suggesting that viral infection did not impact the pharmacokinetics of AVI7288.
11
Figure S6. Maximum Plasma Concentration after Intramuscular and Intravenous
Administration of AVI-7288 in Uninfected Nonhuman Primates
Maximum plasma concentration (Cmax) after intramuscular administration of AVI-7288 (red) was
lower than Cmax after intravenous administration of AVI-7288 (blue) in uninfected nonhuman
primates on day 1. Values depicted are mean ± standard deviation.
12
Table S1. Summary of Studies in Nonhuman Primates
Study
AVI-6003 pilot efficacy 1
AVI-6003 pilot efficacy 2
Determination of effective
AVI-6003 dose
Comparison of AVI-6003 with
individual components
Comparison of AVI-6003 with
AVI-7288
Treatment
Route*
Dose
(mg/kg)
Equivalent
AVI-7288
Dose
(mg/kg)
Saline control
SC + IM
-
0
1
Once daily x 15 d
1M
33
0
AVI-6003
SC + IM
40
20
1
Once daily x 15 d
4M
33
100
Saline control
SC + IP
-
0
1
Once daily x 15 d
1M
28
0
AVI-6003
SC
40
20
1
Once daily x 15 d
1M/2F
28
100
AVI-6003
SC + IP
30
15
1
Once daily x 15 d
3M
28
100
AVI-6003
IV
40
20
1
Once daily x 15 d
3M
28
100
Saline control
IV
-
0
1
Once daily x 15 d
1M
28
0
PMOplus control
IV
30
0
1
Once daily x 15 d
4M
28
0
AVI-6003
IV
7.5
3.75
1
Once daily x 15 d
5M
28
60
AVI-6003
IV
15
7.5
1
Once daily x 15 d
5M
28
60
AVI-6003
IV
30
15
1
Once daily x 15 d
5M
28
80‡
Saline control
IV
-
0
1
Once daily x 15 d
1M
28
0
AVI-7287
IV
7.5
0
1
Once daily x 15 d
5M
28
0
AVI-7288
IV
7.5
7.5
1
Once daily x 15 d
5M
28
60
AVI-6003
IV
15
7.5
1
Once daily x 15 d
5M
28
60
Saline control
IV
-
0
1
Once daily x 15 d
3M/2F
41
0
AVI-6003
IV
30
15
1
Once daily x 15 d
6M/4F
41
90
AVI-7288
IV
15
15
1
Once daily x 15 d
6M/4F
41
100
Group
Treatment
Delay
(h)
Regimen
No. of
Animals†
Duration
of
Follow-up
(days)
Survival
(%)
13
Study
AVI-7288 delayed time to
treatment
AVI-7288 intramuscular
feasibility
Treatment
Route*
Dose
(mg/kg)
Equivalent
AVI-7288
Dose
(mg/kg)
Saline control
IV
-
0
1
Once daily x 14 d
4M/2F
41
0
AVI-7288
IV
15
15
1
Once daily x 14 d
5M/1F
41
83‡
AVI-7288
IV
15
15
24
Once daily x 14 d
5M/1F
41
83
AVI-7288
IV
15
15
48
Once daily x 14 d
5M/1F
41
100
AVI-7288
IV
15
15
96
Once daily x 14 d
5M/1F
41
83
Saline control
IM
-
0
1
Once daily x 14 d
3M/3F
40
0
AVI-7288
IM
7.5
7.5
1
Once daily x 14 d
3M/3F
40
83
AVI-7288
IM
15
15
1
Once daily x 14 d
3M/3F
40
83
AVI-7288
IM
30
30
1
Once daily x 14 d
3M/3F
40
100
Saline control
IV
-
0
1
Once daily x 14 d
3M/3F
39
0
AVI-7288
IV
3.75
3.75
1
Once daily x 14 d
3M/3F
39
0
AVI-7288
IV
7.5
7.5
1
Once daily x 14 d
3M/3F
39
33
AVI-7288
IV
15
15
1
Once daily x 14 d
3M/3F
39
83
AVI-7288
IV
3.75
3.75
N/A
Single dose
3M/3F
N/A
N/A
AVI-7288
IV
7.5
7.5
N/A
Once daily x 14 d
3M/3F
N/A
N/A
AVI-7288
IV
15
15
N/A
Once daily x 14 d
3M/3F
N/A
N/A
Group
Treatment
Delay
(h)
Regimen
No. of
Animals†
Duration
of
Follow-up
(days)
Survival
(%)
Pharmacokinetic studies
AVI-7288 pharmacokinetics
and pharmacodynamics in
infected macaques
AVI-7288 pharmacokinetics in
uninfected macaques§
* IM = intramuscular; IP = intraperitoneal; IV = intravenous; SC = subcutaneous.
† M denotes male; F denotes female.
‡ Death of one animal in group considered to be related to anesthesia, which was required for administration of study drug.
§ As this pharmacokinetic study was completed in uninfected monkeys, the treatment delay, duration of follow-up, and survival are not applicable (N/A).
14
Table S2: Characteristics of Healthy Volunteers in Multiple Ascending Dose Study
AVI-7288 Dose
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
Total
Characteristic
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=40)
Male sex – no. (%)
7 (70)
2 (33)
5 (83)
2 (33)
2 (33)
2 (33)
20 (50)
White
5 (50)
2 (33)
1 (17)
4 (67)
1 (17)
3 (50)
16 (40)
Black
5 (50)
3 (50)
5 (83)
2 (33)
5 (83)
3 (50)
23 (58)
Asian
0
1 (16.7)
0
0
0
0
1 (3)
Mean
32.3
31.2
35.3
31.2
28.7
34.0
32.1
Range
22-47
20-40
26-44
21-49
23-45
18-45
18-49
Mean
86.4
65.5
85.4
73.4
71.8
80.9
78.1
Range
72.0-100.5
55.1-88.5
69.2-98.5
63.1-89.3
58.1-88.0
71.9-92.1
55.1-100.5
Mean
28.7
22.8
27.9
24.3
25.3
26.4
26.2
Range
24.3-33.7
19.9-26.1
24.5-34.1
20.7-32.4
23.3-28.7
21.9-34.2
19.9-34.2
Race – no (%)*
Age – yr
Weight – kg
Body mass index†
* Race was self-reported.
† The body-mass index is the weight in kilograms divided by the square of the height in meters.
15
Table S3. Summary of Adverse Events Occurring during Study Period by Severity
AVI-7288 Dose
All
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
8 (80)
5 (83)
0
2 (33)
2 (33)
6 (100)
15 (50)
23 (58)
Mild
Moderate
Severe
3 (30)
0
0
2 (33)
2 (33)
0
0
0
0
1 (17)
0
0
0
0
0
5 (83)
0
0
8 (27)
2 (7)
0
11 (28)
2 (5)
0
Headache
Mild
Moderate
Severe
2 (20)
0
0
1 (17)
2 (33)
0
0
0
0
1 (17)
0
0
0
0
0
4 (67)
0
0
6 (20)
2 (7)
0
8 (20)
2 (5)
0
Dysgeusia
Mild
Moderate
Severe
1 (10)
0
0
2 (33)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2 (7)
0
0
3 (8)
0
0
Dizziness
Mild
Moderate
Severe
1 (10)
0
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
2 (5)
0
0
Somnolence
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
1 (10)
0
0
1 (17)
1 (17)
0
0
0
0
0
0
0
1 (17)
0
0
1 (17)
1 (17)
0
3 (10)
2 (7)
0
4 (10)
2 (5)
0
Abdominal pain
Mild
Moderate
Severe
0
0
0
1 (17)
1 (17)
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
1 (3)
0
1 (3)
1 (3)
0
Diarrhoea
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
1 (17)
0
1 (3)
1 (3)
0
1 (3)
1 (3)
0
Body System
Preferred Term
Severity*
At least one adverse event
Nervous system disorders
Gastrointestinal disorders
16
AVI-7288 Dose
Body System
All
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
Preferred Term
Severity*
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
Constipation
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Dry mouth
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Mouth cyst
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
Mild
Moderate
Severe
2 (20)
0
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
2 (33)
0
0
3 (10)
0
0
5 (13)
0
0
Infusion site pain
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
2 (5)
0
0
Application site irritation
Mild
Moderate
Severe
0
0
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
2 (3)
0
0
Influenza-like illness
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
Oedema peripheral
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
General disorders and
administration site conditions
17
AVI-7288 Dose
Body System
All
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
Preferred Term
Severity*
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
Pain
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
1 (17)
0
0
0
0
0
1 (17)
0
0
1 (17)
0
0
1 (17)
1 (17)
0
4 (13)
1 (3)
0
4 (10)
1 (3)
0
Flank pain
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (17)
0
0
0
0
0
2 (7)
0
0
2 (5)
0
0
Musculoskeletal pain
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2 (33)
0
0
2 (7)
0
0
2 (5)
0
0
Pain in extremity
Mild
Moderate
Severe
0
0
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
1 (3)
1 (3)
0
1 (3)
1 (3)
0
Mild
Moderate
Severe
2 (20)
1 (10)
0
1 (17)
0
0
0
0
0
1 (17)
0
0
0
0
0
0
0
0
2 (7)
0
0
4 (10)
1 (3)
0
Rash
Mild
Moderate
Severe
2 (20)
0
0
1 (17)
0
0
0
0
0
1 (17)
0
0
0
0
0
0
0
0
2 (7)
0
0
4 (10)
0
0
Dermatitis contact
Mild
Moderate
Severe
0
1 (10)
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
1 (3)
1 (3)
0
Musculoskeletal and connective
tissue disorders
Skin and subcutaneous tissue
disorders
18
AVI-7288 Dose
Body System
All
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
Preferred Term
Severity*
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
Pruritis
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
1 (3)
1 (3)
0
Gastroenteritis
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
Nasopharyngitis
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (17)
0
0
2 (7)
0
0
2 (5)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Infections and infestations
Investigations
Increased blood pressure
Increased urine albumin to
creatinine ratio
Renal and urinary disorders
Micturition disorder
19
AVI-7288 Dose
Reproductive system and breast
disorders
Dysmenorrhea
*
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
Severity*
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Body System
Preferred Term
All
Worst severity in case of multiple occurrences per preferred term.
† All subjects = Subjects who took any study medication (counted only once). Subjects were counted only once in each body system and preferred term.
20
Table S4. Summary of Treatment-Related Adverse Events Occurring during Study Period by Severity
AVI-7288 Dose
All
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
3 (30)
5 (83)
0
1 (17)
1 (17)
3 (50)
10 (33)
13 (33)
Mild
Moderate
Severe
2 (20)
0
0
3 (50)
1 (17)
0
0
0
0
1 (17)
0
0
0
0
0
1 (17)
0
0
5 (17)
1 (3)
0
7 (18)
1 (3)
0
Headache
Mild
Moderate
Severe
2 (20)
0
0
2 (33)
1 (17)
0
0
0
0
1 (17)
0
0
0
0
0
0
0
0
3 (10)
1 (3)
0
5 (13)
2 (3)
0
Dysgeusia
Mild
Moderate
Severe
0
0
0
2 (33)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2 (7)
0
0
2 (5)
0
0
Somnolence
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
2 (33)
0
0
3 (10)
0
0
3 (8)
0
0
Musculoskeletal pain
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2 (33)
0
0
2 (7)
0
0
2 (5)
0
0
Pain in extremity
Mild
Moderate
Severe
0
0
0
1 (17)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Body System
Preferred Term
Severity*
At least one treatment-related
adverse event
Nervous system disorders
Musculoskeletal and connective
tissue disorders
21
AVI-7288 Dose
All
All
Placebo
1 mg/kg
4 mg/kg
8 mg/kg
12 mg/kg
16 mg/kg
AVI-7288
Subjects†
Severity*
(n=10)
(n=6)
(n=6)
(n=6)
(n=6)
(n=6)
(n=30)
(n=40)
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
2 (5)
0
0
Infusion site pain
Mild
Moderate
Severe
1 (10)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
2 (5)
0
0
Peripheral oedema
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Gastrointestinal disorders
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
1 (3)
0
0
1 (3)
0
0
Mild
Moderate
Severe
Severe
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (17)
0
0
0
0
0
0
0
1 (3)
0
0
0
1 (3)
0
0
0
Body System
Preferred Term
General disorders and
administration site conditions
Diarrhoea
Investigations
Increased urine albumin to
creatinine ratio
*
Worst severity in case of multiple occurrences per preferred term.
† All subjects = Subjects who took any study medication (counted only once). Subjects were counted only once in each body system and preferred term.
22
Table S5. Pharmacokinetic Parameters of AVI-7288 in Infected Nonhuman Primates
Mean (SD)
Cmax
(ng/mL)
Tmax
(hr)
AUC0-24
(ng*hr/mL)
Plasma halflife (hr)
ClP
(mL/hr/kg)
Vss
(mL/kg)
3.75 mg/kg*
19,031 (3,531)
0.25 (0.00)
21,826 (4,159)
3.42 (0.24)
178 (42)
886 (272)
7.5 mg/kg
30,480 (9,349)
0.29 (0.10)
39,667 (7,888)
3.67 (0.72)
195 (39)
1,016 (201)
15 mg/kg
66,961 (29,206)
0.25 (0.00)
78,162 (28,993)
3.41 (0.31)
222 (109)
1,092 (551)
3.75 mg/kg
17,364 (2,660)
0.25 (0.00)
22,250 (5,064)
4.14 (1.52)
179 (57)
1,149 (551)
7.5 mg/kg
27,653 (6,955)
0.29 (0.10)
45,452 (8,306)
4.12 (NA)
170 (31)
972 (NA)
15 mg/kg
63,793 (12,677)
0.29 (0.10)
85,923 (17,176)
3.77 (0.84)
180 (34)
974 (316)
3.75 mg/kg
29,144 (7,041)
0.25 (0.00)
39,480 (15,828)
3.66 (0.90)
105 (34)
562 (314)
7.5 mg/kg
35,979 (11,555)
0.25 (0.00)
35,685 (9,282)
4.40 (0.25)
222 (60)
1,431 (402)
15 mg/kg*
65,849 (21,815)
0.21 (0.09)
90,114 (36,565)
3.78 (0.34)
190 (77)
1,018 (366)
3.75 mg/kg
29,336 (11,658)
0.25 (0.00)
46,157 (29,873)
28.5 (NA)
103 (67)
2,291 (NA)
7.5 mg/kg
41,876 (25,611)
0.33 (0.14)
50,261 (30,501)
41.8 (NA)
185 (92)
5,321 (NA)
15 mg/kg
61,923 (8,890)
0.30 (0.11)
76,395 (23,140)
NC (NA)
210 (58)
NC (NA)
Day 1
Day 5
Day 10
Day 14†
AUC0-24 = the area under the plasma concentration-time curve from time 0 to 24 hours after the start of infusion of
study drug; CLp = plasma clearance; Cmax = maximum plasma concentration; NA = SD not applicable, as only one
animal remaining in group; NC = not calculated; SD = standard deviation; Tmax = the time at which Cmax occurs; Vss
= the volume of distribution under steady-state conditions
* Pharmacokinetic parameters from one animal excluded from descriptive statistics because of an anomalous profile.
† Few animals remained on study at day 14 in the lower dose groups.
23
Table S6. Pharmacokinetic Parameters of AVI-7288 in Uninfected Nonhuman Primates
Mean (SD)
Cmax
(ng/mL)
Tmax
(hr)
AUC0-24
(ng*hr/mL)
Plasma halflife (hr)
ClP
(mL/hr/kg)
Vss
(mL/kg)
3.75 mg/kg*
21,247 (13,854)
0.25 (0.00)
35,277 (23,740)
4.03 (1.12)
153 (95)
928 (588)
7.5 mg/kg
24,292 (3,646)
0.25 (0.00)
28,538 (4,719)
4.41 (0.48)
269 (52)
1,735 (500)
15 mg/kg
56,863 (13,327)
0.25 (0.00)
69,680 (12,293)
4.94 (1.23)
230 (37)
1,672 (650)
7.5 mg/kg
30,554 (8,658)
0.25 (0.00)
34,588 (10,112)
6.53 (0.94)
230 (55)
2,177 (622)
15 mg/kg
59,076 (26,353)
0.33 (0.20)
72,456 (16,092)
7.02 (2.09)
215 (45)
2,358 (920)
7.5 mg/kg
28,908 (9,508)
0.25 (0.00)
32,189 (9,807)
8.49 (0.37)
255 (93)
2,398 (410)
15 mg/kg
55,031 (17,968)
0.25 (0.00)
60,108 (18,220)
11.55 (0.78)
268 (77)
5,020 (313)
7.5 mg/kg
26,394 (5,585)
0.25 (0.00)
30,645 (7,409)
NC (NA)
256 (54)
NC (NA)
15 mg/kg
53,406 (12.303)
0.25 (0.00)
62,930 (16,811)
NC (NA)
254 (74)
NC (NA)
Day 1
Day 5
Day 10
Day 14
AUC0-24 = the area under the plasma concentration-time curve from time 0 to 24 hours after the start of infusion of
study drug; CLp = plasma clearance; Cmax = maximum plasma concentration; NA = SD not applicable, as only one
animal remaining in group; NC = not calculated; SD = standard deviation; Tmax = the time at which Cmax occurs; Vss
= the volume of distribution under steady-state conditions
* Animals in the 3.75 mg/kg received only one dose of AVI-7288.
24
Table S7. Pharmacokinetic Parameters of AVI-7288 in Uninfected Humans
Mean (SD)
Cmax
(ng/mL)
Tmax
(hr)
AUC0-24
(ng*hr/mL)
Plasma halflife (hr)
ClP
(mL/hr/kg)
Vss
(mL/kg)
1 mg/kg
6,190 (1,331)
0.45 (0.14)
9,277 (1,844)
3.93 (0.93)
112 (25)
228 (59)
4 mg/kg
15,940 (3,110)
0.67 (0.01)
28,019 (5,365)
4.53 (1.06)
146 (29)
409 (58)
8 mg/kg
37,876 (2,793)
0.66 (0.01)
58,553 (3,311)
4.05 (0.49)
136 (8)
344 (27)
12 mg/kg
51,818 (22,694)
0.92 (0.33)
82,807 (17,583)
3.79 (0.40)
150 (32)
397 (192)
16 mg/kg
56,354 (13,901)
0.72 (0.14)
88,278 (14,486)
3.87 (0.78)
184 (28)
441 (99)
1 mg/kg
4,647 (616)
0.67 (0.00)
8,709 (1,431)
4.49 (1.35)
118 (22)
764 (214)
4 mg/kg
20,723 (4,617)
0.67 (0.00)
39,011 (9,909)
5.62 (1.56)
108 (27)
881 (95)
8 mg/kg
37,068 (4,476)
0.67 (0.00)
69,356 (4,169)
3.68 (0.85)
116 (7)
670 (65)
12 mg/kg
35,439 (4,346)
0.73 (0.15)
84,697 (10,046)
3.50 (1.02)
143 (16)
1,200 (485)
16 mg/kg
69,358 (14,525)
0.67 (0.02)
130,036 (36,974)
3.99 (0.55)
132 (38)
882 (430)
Day 1
Day 14
AUC0-24 = the area under the plasma concentration-time curve from time 0 to 24 hours after the start of infusion of
study drug; CLp = plasma clearance; Cmax = maximum plasma concentration; SD = standard deviation; Tmax = the
time at which Cmax occurs; Vss = the volume of distribution under steady-state conditions
25
Table S8. Median, Geometric Mean, and 5th and 95th Percentile Values from 10,000
Simulation AUC0-inf Values in Infected Nonhuman Primates and Uninfected Humans
Parameter
Infected
Nonhuman
Primates
(15 mg/kg)
Infected
Humans
(9.6 mg/kg)
Infected
Humans
(10 mg/kg)
Infected
Humans
(11 mg/kg)
Infected
Humans
(15 mg/kg)
Median
75,800
70,300
73,300
80,600
109,900
Geometric mean
82,100
70,600
73,600
80,900
110,300
5th percentile
48,200
57,800
60,300
66,300
90,400
170,000
89,700
93,400
103,000
140,000
th
95 percentile
AUC0-inf = the area under the plasma concentration-time curve from time 0 to infinity. Results are presented as
ng*hr/mL.
26