Immunogenicity And Safety Of An Alum-Adjuvanted Inactivated H7N9 Influenza Vaccine
Phase: PHASE1
Conditions: H7N9 Influenza
Sponsor: Shanghai Institute Of Biological Products

SIBP 
Protocol ID: 2016L09902/3 
1/13 
 
 
 
 
CLINICAL TRAIL PROTOCOL 
 
 
Protocol Title: Immunogenicity and Safety of an Alum-adjuvanted 
Inactivated H7N9 Influenza Vaccine: a Randomized, Blind, 
Placebo-controlled, a Phase I/II Clinical Trial. 
 
 
Collaborators: Henan Center for Disease Control and Prevention 
 
Sponsor: Shanghai Institute of Biological Products 
 
 
 
 
 
 
 
Document Date: October 17, 2017 
 

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Protocol ID: 2016L09902/3 
2/13 
 
BACKGROUD INFORMATION  
The first human infection with novel avian-origin influenza A (H7N9) was reported in China 
in March 2013. As of 26th October 2017, a total of 1564 laboratory-confirmed human 
H7N9 infectious, including at least 612 deaths, with high mortality (approximately 40%), 
have been reported to World Health Organization (WHO) since early 2013. Recently, the 
activity of the H7N9 virus appears increasing trend. 
 
Despite the fact that the H7N9 virus has not yet been able to sustain human to human 
transmission in the current, it is difficult to judge its epidemic trend. A lot of researches 
demonstrates that the novel H7N9 the viruses have been evolving from low pathogenic 
into highly pathogenic and process a potential pandemic threat to public health worldwide 
due to lack of pre-existing immunity in humans. Therefore, it is prudent to develop 
strategies to prepare for H7N9 pandemic outbreak. Vaccination is currently considered as 
effective strategy of controlling influenza viruses with pandemic potential. 
 
In 2013 May, three vaccine strains, A/Shanghai/2/2013 (IDCDC-RG32A, NIBRG-267) and 
A/Anhui/1/2013 (NIBRG-268), were used for vaccine development. Shanghai Institute of 
Biological Products (SIBP) carried out research and development of H7N9 avian influenza 
virus vaccine. The whole and split virus inactivated vaccines were taken as the research 
direction of the traditional influenza vaccine strategy. However, previous studies indicated 
the H7-containing vaccines were poor immunogenic in humans and difficult to induce 
effectively protective antibodies. Adjuvantation is universally regarded as effective means 
of immune-enhancing and antigen-sparing. As aluminum is the only approved adjuvant in 
China, we added the aluminum hydroxide adjuvant in the vaccine, to enhance the immune 
response. At present, vaccine produced by SIBP has received clinical approval, ready to 
carry out phase I/II clinical trials, safety and immunogenicity of the vaccine will be 
evaluated by clinical trials. 
 
STUDY OBJECT 
First objects: to evaluate the safety of different doses (7.5 μg, 15 μg and 30 μg HA antigen) 
of influenza A (H7N9) virus inactivated vaccine in healthy population aged 12 years and 
over, according to investigate occurrence of adverse events for 30 days after two 
inoculations. 
 
Secondary objects: to evaluate the immunogenicity of different doses (7.5 μg, 15 μg and 
30 μg HA antigen) of influenza A (H7N9) virus inactivated vaccine in healthy population 

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aged 12 years and over, according to investigate antibodies at day 21 after two 
inoculations. 
 
STUDY DESIGN 
This will be a randomized, blind, one-center, placebo control study conducted by Henan 
Center for Disease Control and Prevention. There will be two phases. The vaccine dosage 
and the immunization procedures of phase II will be determined according to the safety 
and immunogenicity results of phase I.  
 
There will be three dosage groups of H7N9 influenza virus inactivated whole virus vaccine: 
Group A (7.5 μg HA antigen H7N9 influenza vaccine); Group B (15 μg HA antigen H7N9 
influenza vaccine) and Group C (30 μg HA antigen H7N9 influenza vaccine). Every group 
is divided into subgroups according to different age. Subgroup 1 (group of adults aged 18 
years and above); Subgroup 2 (group of youth aged 12~17 years). Each subgroup will be 
treated with 2 placebo controls (Aluminum Hydroxide and PBS).  
 
All subjects will be inoculated with the corresponding vaccine at 0 and 21 days. Then 
subjects will receive immediate response observation for 30 minutes and systematic 
safety observation for 7 days after each dose of vaccination. After 7 days of vaccination, 
the adverse events will be observed by weekly regular follow-up and the subjects' 
reporting. 
 
All subjects will be collected serum samples at before the first dose of vaccination, day 21 
after the first dose of vaccination and day 21 after the second dose of vaccination. All 
serum samples will be detected HI antibody. See Table below. 
 
Group 
Age 
Dosage and Number of subjects 
Inoculation 
Serum 
collection 
Safety 
observation 
A1 
≥18 years 
7.5 μg per dose 
30 
Day 0,21 
(+7 days)  
Day 0,21 
(+7),42(+10) 
The local and 
systemic AE and 
SAE will be 
observed for 6 
months after 
inoculation. 
Placebo (Alum) 
15 
Placebo (PBS) 
15 
A2 
12~17 years 
7.5 μg per dose 
30 
Placebo (Alum) 
15 
Placebo (PBS) 
15 
B1 
≥18 years 
15 μg per dose 
30 
Placebo (Alum) 
15 
Placebo (PBS) 
15 
B2 
12~17 years 
15 μg per dose 
30 
Placebo (Alum) 
15 
Placebo (PBS) 
15 
C1 
≥18 years 
30 μg per dose 
30 
Placebo (Alum) 
15 

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Placebo (PBS) 
15 
C2 
12~17 years 
30 μg per dose 
30 
Placebo (Alum) 
15 
Placebo (PBS) 
15 
Total 
6 subgroups 
/ 
360 
/ 
/ 
/ 
 
STUDY ENDPOINT 
Primary endpoint 
To investigate adverse events (AE) and serious adverse events (SAE) within 30 days after 
the first and second inoculation as primary endpoint. 
 
Secondary endpoint 
To detect the serum HI antibody at day 21 after first and second inoculation as secondary 
endpoint. 
 
SAFETY OBSERVATION 
Systemic adverse reactions: fever, headache, fatigue, fatigue, nausea, vomiting, diarrhea, 
muscle pain, cough, allergy (anaphylactic shock, urticaria, vascular edema, etc.). 
 
Local adverse reactions: pain, redness, swelling, induration, rash (injection site), itching, 
skin mucous membrane. 
 
The events (hospitalization, hospitalization time, disability of work ability, life or death, 
congenital malformation and so on) occur during clinical trial. 
 
IMMUNOGENICITY OBSERVATION 
Seroconversion: 1:10 is the lowest serum dilution. Prevaccination HI titer < 1:10 and 
postvaccination titer ≥ 1:40 or a prevaccination titer ≥ 1:10 and at least a 4-fold increase in 
the postvaccination titer 
 
Seroprotection: Postvaccination titer ≥ 1:40 is considered to have antibody protection. 
 
GMT: geometric mean titer. 
 
GMI: GMT increase fold. 
 
EVALUATION CRITERIA OF SAFETY 
The safety evaluation of each group will be divided into three stages. From the first dose 

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inoculation to day 7 after inoculation; From the first dose inoculation to day 21 after 
inoculation; From the second dose inoculation to day 30 after the second dose inoculation. 
The first stage is the basis for the follow-up group of the same vaccine, and will be carried 
out under blind. The latter two stages are the basis for evaluation of vaccine safety, and 
will be carried out under unblind. If each evaluation is not found level 4 local and systemic 
adverse reactions associated with vaccine and the total incidence rate of level 3 of local 
and systemic adverse reactions and abnormal laboratory test which associated with the 
vaccination is lower than 15%, that the vaccine will be acceptable safety. 
 
EVALUATION CRITERIA OF IMMUNOGENICITY 
If seroconversion rate is higher than 40%, the antibody protection rate higher than 70%, 
GMI more than 2.5 times, the vaccine will be acceptable immunogenicity. 
 
GROUP PROCESS 
In order to protect the rights of the subjects, each group will be carried out after evaluating 
the safety of last group. The sequence of inoculation is from low dosage to high, from 
older age to young. The flow sequence is shown in table below. 
Day 
Group A 
Group B 
Group C 
0 
Group A1  First Dose 
 
 
8 
Group A2  First Dose 
Group B1  First Dose 
 
16 
 
Group B2  First Dose 
Group C1  First Dose 
21 
Group A1  Second Dose 
 
 
24 
 
 
Group C2  First Dose 
29 
Group A2  Second Dose 
Group B1  Second Dose 
 
37 
 
Group B2  Second Dose 
Group C1  Second Dose 
45 
 
 
Group C2  Second Dose 
51 
Complete 30 days 
observation of Group A1 
 
 
59 
Complete 30 days 
observation of Group A2 
Complete 30 days 
observation of Group B1 
 
67 
 
Complete 30 days 
observation of Group B2 
Complete 30 days 
observation of Group C1 
75 
 
 
Complete 30 days 
observation of Group C2 
201 
Complete 180 days 
 
 

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observation of Group A1 
(collect SAE) 
209 
Complete 180 days 
observation of Group A2 
(collect SAE) 
Complete 180 days 
observation of Group B1 
(collect SAE) 
 
217 
 
Complete 180 days 
observation of Group B2 
(collect SAE) 
Complete 180 days 
observation of Group C1 
(collect SAE) 
225 
 
 
Complete 180 days 
observation of Group C2 
(collect SAE) 
 
STUDY STATUS 
Record Verification: October 2017 
Study Start: November 2017 
Primary Completion: October 2018 
Study Completion: December 2019 
 
 
ELIGIBILITY 
Inclusion Criteria: 
Over the age of 12 years, healthy population 
Subjects/ (and the guardian) informed consent, voluntarily participated and signed the 
informed consent form, with the ability to use thermometers, scales and to fill in diary 
cards as required 
To comply with the requirements of clinical trial program, receive blood test before and 
after immunization and cooperate with follow-up 
 
Exclusion Criteria: 
A history of influenza A (H7N9) virus infection or suspected infection 
Abnormal blood routine, blood biochemistry and urine routine examination indexes 
Allergy to any component in the vaccine (allergy history of any previous vaccination), 
especially for egg allergy 
History of asthma, history of thyroid resection, vascular nerve edema, diabetes mellitus, 
and hypertension cannot be controlled by medicine, liver and kidney diseases or 
malignant tumor history 

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Suffered from any serious illness, such as cancer, autoimmune disease, progressive 
atherosclerotic disease or complications of diabetes, chronic obstructive pulmonary 
disease, need oxygen therapy for acute or progressive liver or kidney disease, congestive 
heart-failure etc. 
History of signs disease or symptoms of neurological symptoms 
Suffering from severe chronic diseases (such as Down's syndrome, diabetes, sicklemia or 
neurological disorders, Green's Barre syndrome) 
Acute attacks of various acute or chronic diseases in the past 7 days 
Known or suspected of respiratory disease, acute infection or chronic disease active 
period, HIV infection, cardiovascular disease, severe hypertension, malignant tumor 
during treatment, skin diseases 
Congenital malformations or developmental disorders, genetic defects, severe 
malnutrition etc. 
No spleen, functional absence of spleen, and splenectomy or splenectomy without any 
condition 
Autoimmune diseases or immunodeficiency have been treated with immunosuppressive 
agents in the past 6 months 
History of epilepsy, convulsions, or a family history of psychosis 
Abnormal coagulation function (such as coagulation factor deficiency, coagulation 
disorders, platelet abnormalities), or obvious bruising or coagulopathy 
The blood products were received within 3 months prior to the acceptance of the vaccine 
Received a live vaccine within 14 days prior to receiving the vaccine, or received a subunit 
or inactivated vaccine within 7 days 
Fever within 3 days prior to vaccination, axillary temperature ≥38℃ 
Fever When inoculating vaccine, axillary temperature >37.0℃ 
Women are pregnant or in the near future planned pregnancy or pregnancy test positive 
Participants in another clinical trial 
The researchers believe that there may be any impact on the assessment of the trial. 
 
DATA STATISTICS 
Statistics analysis of immunogenicity  
The prevaccination and postvaccination geometric mean of antibody and its 95% 
confidence interval (confidence interval, CI) will be described. The postvaccination serum 
conversion rate and protection rate of antibody and their 95% CI will be described. 
 
Comparisons will be conducted to evaluate differences in response between study groups 

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using a 
2

 test or Fisher's exact test to compare to the difference of serum conversion 
rate and protection rate. Statistical significance will be considered at a level of α = 0.05 
and all tests will be 2-sided. 
 
If serum HI antibody conversion rate is more than 40%; protection rate more than 70%; 
GMI more than 2.5 folds, the immunogenicity will reach the design requirements.  
 
Statistics analysis of safety 
After the first dose inoculation and the second dose inoculation, the number of adverse 
reactions (rate), number of cases and adverse reaction grade were recorded. 
The number of adverse events after inoculation will be described. 
 
Comparisons will be conducted to evaluate differences in response between study groups 
using a 
2

 test or Fisher's exact test to compare to the difference of adverse events rate. 
Statistical significance will be considered at a level of α = 0.05 and all tests will be 
2-sided. 
 
Comparison of compliance 
The rate of expulsion and the rate of drug combination will be described. 
 
Comparisons will be conducted to evaluate differences in response between study groups 
using a 
2

 test or Fisher's exact test to compare to the difference of the rate of 
expulsion and the rate of drug combination. Statistical significance will be considered at a 
level of α = 0.05 and all tests will be 2-sided. 
 
 

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APPENDIX Ⅰ. GRADING STANDARDS FOR SEVERITY OF 
SAFETY INFORMATION 
The Event of clinical response and laboratory abnormalities after vaccination is judged by 
the China State Food and Drug Administration on "Guidelines for the classification of 
adverse events in vaccine clinical trials ". 
Grading of Local Adverse Events 
Local 
Adverse 
Reactions 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Potential Life 
Threatened 
(Grade 4) 
Pain 
Do not affect 
activity 
Influence 
activities 
or 
multiple use of 
non-narcotic 
analgesics 
Affect daily activities 
or multiple use of 
narcotic analgesics 
Emergency 
or 
hospitalization 
Induration 
<15 mm 
15-30 mm 
>30 mm 
Gangrene 
or 
exfoliative 
dermatitis 
Redness 
<15 mm 
15-30 mm 
>30 mm 
Gangrene 
or 
exfoliative 
dermatitis 
Swelling 
<15 mm and 
does 
not 
affect activity 
15-30 
mm 
or 
affect activity 
>30 mm or limit daily 
activity 
Gangrene 
Rash 
(injection 
site) 
<15 mm 
15-30 mm 
>30 mm 
 
Itching 
Injection site 
itching 
Injection 
of 
moderate itching 
Overall Itching 
 
Mucocutan
eous 
Redness 
Diffuse, papular 
rash, 
dry, 
desquamation 
Blister, 
damp, 
desquamation or ulcer 
Peeling 
dermatitis, 
involving 
mucous 
membranes, or 
erythema 
multiforme, 
or 
suspected 

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Stevens-Johnso
ns syndrome 
 
Grading of Systemic Adverse Events 
Systemic 
Adverse 
Reactions 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Potential Life 
Threatened 
(Grade 4) 
Fever 
37.1-37.5℃ 
37.6-39.0℃ 
>39.0℃ 
 
Headache 
No activity, no 
treatment 
Transient, slightly 
affected activity 
requiring 
treatment 
(multiple use of 
non-narcotic 
analgesics) 
Severe effects on 
daily activities, 
initial anesthetic 
response 
Refractory, 
repeated 
anesthetic 
treatment. 
Emergency or 
hospitalization 
Fatigue and 
fatigue 
The normal 
activity is less 
than 48 hours, 
and it did not 
affect the activity 
Normal activity 
decreased from 
20% to 50% > 48 
hours, slightly 
affecting activities 
Normal activity 
decreased by 
more than 50%, 
seriously affecting 
daily activities, 
unable to work 
Unable to take 
care of oneself, 
emergency or 
hospitalization 
Nausea and 
vomiting 
1~2 times per 24 
hours, intake is 
normal and does 
not affect the 
activity 
2 ~ 5 times per24 
hours, intake is 
significantly 
reduced, or 
limited activity 
> 6 times within 24 
hours, no obvious 
intake, the need for 
intravenous infusion
requires 
hospitalization or 
other nutrition 
because of 
hypotension, 
shock 
Diarrhea 
Mild or transient, 
2 ~ 3 stools per 
day or mild 
diarrhea lasting 
for less than 1 
weeks 
Moderate or 
persistent, 4~5 
times per day, or 
more than 1 week 
diarrhea 
> 6 times of water 
stool per day, or 
blood diarrhea, 
orthostatic 
hypotension 
Hypotension 
shock requiring 
hospitalization 

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Myalgia 
Not affecting daily 
activities 
Muscle 
tenderness at the 
non injection site 
slightly affects 
daily activity 
Severe muscle 
tenderness 
seriously affects 
daily activities  
The symptoms 
are obvious, 
muscle necrosis, 
emergency or 
hospitalization 
Cough 
Transient, without 
treatment 
Persistent cough, 
effective 
treatment 
Paroxysmal 
cough, treatment 
can not control 
Emergency or 
hospitalization 
Allergy 
Pruritus without 
skin rash 
Local urticaria 
Extensive urticaria, 
vascular edema 
Severe allergy 
Other 
adverse or 
clinical 
adverse 
reactions 
(based on 
the 
correspondi
ng criteria) 
Do not affect 
activities 
Slightly affects 
activities without 
drug treatment 
Serious impact on 
daily activities 
requires drug 
treatment 
 
 
Grading of Blood Biochemical 
Serum 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Potential Life 
Threatened 
(Grade 4) 
Elevated liver 
function caused by 
AST ALT, influence 
factors 
1.25-2.5 *ULN 
2.6-5*ULN 
5.1-10*ULN 
>10*ULN 
Creatinine 
1.1-1.5*ULN 
1.6-3.0*ULN 
3.1-6*ULN 
>6*ULN 
Urea nitrogen (BUN) 1.25-2.5 *ULN 
2.6-5*ULN 
5.1-10*ULN 
>10*ULN 
Bilirubin: elevation 
caused by factors, 
but functional 
examination is 
1.1-1.5*ULN 
1.6-2.0*ULN 
2.0-3.0*ULN 
>3.0*ULN 

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normal 
Bilirubin: the 
increase caused by 
the factors 
associated with the 
increase of liver 
function test index 
1.1-1.25*ULN 
1.26-1.5*ULN 
1.51-1.75*ULN 
>1.75*ULN 
 
Grading of Blood Routine 
Blood 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Potential Life 
Threatened 
(Grade 4) 
Leukocyte 
high(cells/mm3) 
>13000 
13,000-15,000 
15,000-30,000 
>30,000 
Leukocyte low 
(cells/mm3) 
2500-3500 
1500-2499 
1000-1499 
<1000 
 
 
 
Grading of Urine Routine 
Urine 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Potential Life 
Threatened 
(Grade 4) 
Protein 
trace 
1+ 
2+ 
>2+ 
Urine sugar 
trace 
1+ 
2+ 
>2+ 
 
 

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APPENDIX Ⅱ. THE RELATIONSHIP BETWEEN ADVERSE 
EVENTS AND TRIAL VACCINES 
 
Absolutely unrelated: Because of other factors lead to adverse events, there 
is evidence that adverse events are caused by other causes, not related to 
vaccination. 
 
Possible unrelated: Adverse events probable be caused by other factors, 
such as the clinical status of the subjects, other treatment or concomitant 
medication, inconsistent with the known adverse reactions of vaccination. 
 
Quite possible related: Adverse events are consistent with known 
information of vaccine, and there is a causal relationship with the vaccine, not 
by other factors, such as the clinical status of the subjects, or other treatment 
with medication. 
 
Possible related: Adverse events are consistent with known information of 
vaccine, and there is a causal relationship with the vaccine, not by other 
factors, such as the clinical status of the subjects, or other treatment with 
medication. 
 
Related: Adverse events are consistent with known information of vaccine, 
and there is a causal relationship with the vaccine, not by other factors, such 
as the clinical status of the subjects, or other treatment with medication. In 
addition, adverse events will be repeated when subjects are tested with the 
vaccine.