The Development of a COVID19 Oral Vaccine Consisting of Bacillus Subtilis Spores Expressing and Displaying the Receptor Binding Domain of Spike Protein of SARS-COV2
Phase: NA
Conditions: COVID-19 Pneumonia
Sponsor: DreamTec Research Limited

Study Title: The Development of a COVID19 Oral Vaccine Consisting of Bacillus subtilis Spores 
Study sponsor: DreamTec Research Limited 
Principal Investigator 
Name: Kwong Wai Yeung 
Phone: +852 95184614 
Email: Keithkwong@dreamtec.hk 
Date: 1 June 2021 

 
 
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The Development of a COVID19 Oral Vaccine Consisting of 
Bacillus subtilis Spores 
study protocol 
 
 
 
 
Common name of the 
vaccine under test 
Bacillus subtilis 
spore 
Name of the test agent 
 
Vaccine Registration 
Applicants 
main investigators 
 
（Signature） 
Head of study Unit 
（Unit seal） 
Unit of statistical 
analysis 
Contract Research 
Organization 
Application Contact 
Person 
Contact number 
Contact address 
Start and end date of the 
test 
The location of the 
original data 

 
 
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Contents 
Summary ....................................................................................................................................... 4 
1. Test subject .................................................................................................................... 4 
2. Test objective ................................................................................................................. 4 
3. Test vaccine ................................................................................................................... 4 
4. Overall design of test ..................................................................................................... 4 
5. Test population .............................................................................................................. 4 
6. Test schedule .................................................................................................................. 4 
Main body ..................................................................................................................................... 5 
1. Introduction .................................................................................................................... 5 
2. Test subject .................................................................................................................... 8 
3. Test objective  ................................................................................................................ 8 
4. Overall design and arrangement of the test .................................................................... 8 
4.1. 
Test design ......................................................................................................... 8 
4.2. 
Dose, method and course of treatment .............................................................. 8 
5. Subject selection ............................................................................................................ 9 
5.1. 
Inclusion criteria ................................................................................................ 9 
5.2. 
Exclusion criteria  .............................................................................................. 9 
5.3. 
Case withdrawal criteria .................................................................................. 10 
5.4. 
Management of cases of loss  ......................................................................... 10 
6. Test vaccine ................................................................................................................. 10 
6.1. 
Test vaccine ..................................................................................................... 10 
6.2. 
Control tablet ................................................................................................... 11 
6.3. 
Vaccine packaging ........................................................................................... 11 
6.4. 
Administration ................................................................................................. 11 
6.5. 
Vaccine management ....................................................................................... 11 
6.6. 
Combination of medicines use and prohibited medicines ............................... 11 
7. Clinical efficacy indexes and safety evaluation criteria ............................................... 12 
7.1. 
Main efficacy indicators .................................................................................. 12 
7.2. 
Other efficacy indicators ................................................................................. 12 
7.3. 
Security indicators ........................................................................................... 12 
7.4. 
Follow-up examination .................................................................................... 13 
8. Blinding and randomization ......................................................................................... 13 
8.1. 
Blind editing, answer to the blind preservation ............................................... 13 
8.2. 
Emergency unblinding ..................................................................................... 13 
8.3. 
Unblinding rules .............................................................................................. 14 

 
 
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9. Observation of side effect ............................................................................................ 14 
9.1. 
Definition ......................................................................................................... 14 
9.2. 
Criteria for determining the severity of side effect .......................................... 14 
9.3. 
Criteria for determining the relationship between side effect and the study  
vaccine....................... ..................................................................................... 15 
9.4. 
Determination of serious side effect ................................................................ 16 
9.5. 
Report of serious side effect ............................................................................ 16 
10. Data administration ...................................................................................................... 16 
10.1. 
Filing and handing over of case report form ................................................. 16 
10.2. 
Data entry and modification .......................................................................... 17 
10.3. 
Data locked .................................................................................................... 17 
11. Statistic analysis ........................................................................................................... 17 
11.1. 
Analysis data set ............................................................................................ 17 
11.2. 
Statistical analysis method ............................................................................ 17 
12. Quality control and assurance ...................................................................................... 19 
12.1. 
Quality control ............................................................................................... 19 
12.2. 
Quality guarantee .......................................................................................... 20 
13. Medical ethics and related legal issues ........................................................................ 20 
13.1. 
Guidelines for clinical trial on vaccines ........................................................ 20 
13.2. 
Investigator responsibility ............................................................................. 20 
13.3. 
Study subjects and informed consent ............................................................ 20 
13.4. 
Implementation and modification of the study plan ...................................... 21 
13.5. 
Termination of study ..................................................................................... 21 
13.6. 
Relevant legal liability................................................................................... 21 
14. Summary of test report................................................................................................. 21 
15. Data save ...................................................................................................................... 21 
16. Publication and use of study results ............................................................................. 22 
17. Test plan ....................................................................................................................... 22 
Scheme signature page ................................................................................................................ 23 

 
 
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Summary 
1. 
Test subject 
 
efficacy and safety of B. subtilis spore 
2. 
Test objective 
The efficacy and safety of B. subtilis spore were evaluated with simulated oral tablet as 
control 
3. 
Test vaccine 
Test group：Oral approach, 1 capsule of B. subtilis spore once a day. No other medication. 
Start in the morning on an empty stomach. 
 
 
4. 
Overall design of test 
This study was conducted in a single arm design. 
 
5. 
Test population 
Normal healthy people without disease. Excluded those with basic diseases such as hypertension, 
diabetes and hyperglycaemia. Test schedule 
2021.4.01 – 2021.9.30

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Main body 
1.
Introduction
COVID-19 caused by severe acute respiratory syndrome corona virus 2 (SARS-CoV-2). By
December 2020, it had infected more than 80 million people and killed more than 1.7 million 
people all over the world. Due to the lack of effective treatment, the development and use of 
COVID-19 vaccine had become an important strategy to control the novel pneumonia epidemic. 
Covid-19 was a single strand plus RNA virus, whose exposed spike protein (S protein) and 
receptor binding domain (RBD) on S protein were the main targets for the design of Covid-19 
vaccines. In March 2020, the Covid-19 vaccine developed by China based on adenovirus vector 
and the mRNA technology platform developed by the United States which were the first vaccines 
entering clinical trials, followed were DNA vaccine and inactivated vaccine. By the end of 
December 2020, there were inactivated vaccine, nucleic acid vaccine (DNA vaccine and mRNA 
vaccine), carrier vaccine, protein subunit vaccine, live attenuated vaccine, virus-like particles 
vaccine. The sixty vaccine candidates in six different technical routes were approved to enter 
clinical trials. Some vaccines had been approved for conditional marketing or emergency use. 
Due to the different development technology routes of different COVID-19 vaccines, there 
were still different shortcomings in terms of safety, effectiveness, stability of transportation and 
storage, and capacity. 
Although the safety of COVID-19 vaccines in phase III clinical trials had been reported to 
be good, there were still theoretical safety concerns. For example, live attenuated vaccines needed 
to replicate in the body and carry a risk of virulent atavism infection or transmission. DNA 
vaccines carried the risk of foreign DNA entering the body and integrating into the host genome, 
resulting in oncogene activation, or inactivation of tumor suppressor genes, or chromosomal 
instability. The synthetic materials and coating materials used in the synthesis of mRNA vaccines 
may be toxic and might cause apoptosis of the surrounding host cells. 
In the introduction of the target product of the Covid-19, WHO proposed that the protection 
efficacy of the Covid-19 should be at least 50%. Among the four vaccines whose effectiveness 
had been announced, the inactivated vaccine had a protective efficacy of 79.34%, the carrier 
vaccine 62-90%, and the mRNA vaccine above 90%, all of which met the requirements. But in 
the context of COVID - 19 pandemic disease, unfavorable evaluation or  
directly compare the clinical efficacy of vaccine were appeared. That was because different 
vaccine clinical trial design and research field, the research results were different. Vaccine real 
protection effect still need to be extensively studied through subsequent large-scale phase IV 

 
 
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clinical trials or epidemiological studies after vaccinating. 
In terms of storage and transportation of the Covid-19, mRNA vaccine required high storage 
conditions due to its unstable and easily degradable. A mRNA vaccine developed by the United 
States and Germany needed to be stored at -70℃.  After thawing, vaccine vials could only be 
stored for 5 days in cold storage (2-8℃). The other mRNA vaccine was stable for only 30 days 
at 2-8 ° C and needed to be stored at -20 ° C. 
Due to the need for virus culture process, the production of inactivated and attenuated 
vaccines required high bio-safety level and long production cycle, so there was some difficulty in 
scaling up production capacity. 
This product was bacillus subtis spore vaccine. Various types of vaccines such as mRNA, 
adenovirus and inactivated virus via injection route were developed to prevent the infection of 
SARS-CoV-2. Although some of them have already been approved to against the COVID-19 
pandemics, various drawbacks including severe side effects and instability in storage may hinder 
their applications. Bacillus subtilis (B. subtilis) is a generally recognized as safe (GRAS) and 
endotoxin-free gram-positive bacterium, which has been extensively employed as a host for 
expression of recombinant proteins. Its dormant spores are extraordinarily resistance to harsh 
environment conditions in gastrointestinal tract. This feature makes it an ideal candidate for oral 
administration to resist the environment. In this study, an engineered B. subtilis spore expressing 
the spike protein receptor binding domain (sRBD) of the SARS-CoV-2 on the surface was 
developed, and the result proved that it successfully increased the neutralizing antibody against 
sRBD in unvaccinated mice and human individuals after oral administration. These findings may 
enable the use of B. subtilis spore as oral vaccine against COVID-19 in the future.  
The oral vaccine presents antigen through the gastrointestinal mucosa and could produce an 
immune response similar to that of conventional injection through different immune pathways. 
With the development of medicine and molecular biology technology, great progress had been 
made in the study of oral vaccine in the presentation of mucosal antigen, which could effectively 
induce immune response and immune tolerance in the body. The toxicity of the vaccine was very 
low, and the LD50 could not be measured by oral or peritoneal route. According to the toxicity 
grading standard, it was basically non-toxic. Long-term toxicity, specific toxicity and 
reproductive toxicity studies did not show any biological damage. Experimental data showed 
that the vaccine was extremely safe and well tolerated by patients. 
 
 
 

 
 
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2. 
Test subject 
efficacy and safety of Bacillus subtilis spore. 
3. 
Test objective 
The efficacy and safety of " Bacillus subtilis spore " were evaluated with simulated tablet 
as control 
4. 
Overall design and arrangement of the test 
4.1. 
Test design 
This study adopted a one-arm experimental design. The whole study mainly collected 
qualified normal healthy people without disease from the hospital, and excluded people with 
basic diseases such as hypertension, diabetes, hyperlipidemia and so on. Once confirmed, they 
were assigned to administer 1 dose of Bacillus subtilis spore. Follow-up observation and 
efficacy examination were performed on days 7, 14, 21 and 28. 
 
If subjects had significant laboratory abnormalities or side effect at the end of the study,  
they should be followed up to normal. 
 
 
4.2. 
Dose, method and course of treatment 
 
Group 
Dosage 
Usage 
Survey 
Test group 
1 Bacillus subtilis spore capsule at a 
time 
Oral administration, once 
28 days 
 
Vaccine time window: Oral administration of the vaccine would be given immediately after the 
subjects were selected. 
 
Essential medicines: over-the-counter medicines such as vitamins and minerals were allowed. 
 

 
 
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Medicines prohibited: antibiotics, thrombolytic drugs, anticoagulants, insulin, β-blockers, 
adenosine preparations, leucocyte adhesion inhibitors, other antioxidants, free radical 
scavengers, etc. 
5. 
Subject selection 
5.1. 
Inclusion criteria 
 
Outpatient service (medical records of outpatient service shall be established), male or  
female, from 18 years old to 60 years old; 
No hypertension, diabetes, hyperlipidemia and other basic diseases; 
 
If the patient was a woman of gestational age, the pregnancy test at the time of screening  
should be negative; Contraception (oral or injectable contraceptives) should be used  
throughout the trial; 
 
Obtained the informed written consent of the subject or the subject's immediate family  
member or the subject's guardian; 
 
There was no serious cardiopulmonary liver and kidney insufficiency. 
 
5.2. 
Exclusion criteria 
Patients under 18 years of age or over 60 years of age; 
 
Dementia who could not cooperate, or affecting the judgment of curative effect; 
 
Women who were pregnant, breastfeeding or who might become pregnant during the  
study and who were not using effective contraceptives; 
 
Patients suffering from tuberculosis, tumor and other organic lesions; 
 
Known alcohol and drug dependence; 
 
To determine the patients who were not suitable for this clinical observation; 
 
People with coma, bleeding tendency and allergic constitution were excluded; 
 
Treatments such as hormones, diuretics, hypertonic dehydrating agents, thrombolysis,  
anti-coagulation, and fibrillation that might affect the results of the study; 
 
Staff directly involved in the study; 
 
 
Other circumstances that other physicians or researchers had judged inappropriate for  
participating in the clinical trial

 
 
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5.3. 
Case withdrawal criteria 
Discontinuation of Bacillus subtilis spore due to side effect determined by the investigator; 
 
Discontinuation of Bacillus subtilis spore due to other reasons (including voluntary withdrawal); 
 
Unable to cooperate; 
For reasons other than research-related, the investigator did not consider it appropriate to 
continue the clinical trial. 
5.4. 
Management of cases of loss 
Definition of cases of loss: all subjects who had completed the informed consent form 
and were screened for entry into the study, at any time and for any reason withdrawal, as 
long as the subjects did not complete the observation period specified in the protocol, 
was called cases of loss, including the following conditions: 
 
A serious side effect occurred in the subject and, in the judgment of the physician, the  
case should be discontinued. 
 
During the study, the subjects' original condition worsened or other symptoms occurred 
during the study that affected the observation of the study, according to the doctors 
decided that the clinical trial should be stopped. 
 
Significant deviations occurred during the implementation of the clinical trial protocol, 
making it difficult to evaluate the vaccine effect. 
 
Subjects were unwilling to continue the clinical trial. 
 
Treatment of cases of loss: contacted subjects with reasons, recorded the time of last 
observation, and completed the assessment items that could be completed. For patients 
who withdrew from the study due to allergy or other side effect and failure to respond to 
treatment, the investigator should take appropriate treatment measures according to the 
actual situation of the subjects. The researchers should properly preserved the relevant 
experimental data of the cases of loss, not only for filing, but also for carrying out the 
complete analysis set statistics. 
 
6. 
Test subject 
6.1. 
Test subject 
 
Generic name: Bacillus subtilis spore 
Specifications: Tablets 
Batch number: 
Storage: under 4°C, sealed in the dark place 
Production unit:  
 
6.2. 
Vaccine packaging 

 
 
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Single 1 capsule, individually packed. 
6.3. 
Administration 
Test group：Oral approach, 1 capsule B. subtilis spore once a day. No other medication. 
Start in the morning on an empty stomach. 
 
Control group：Oral approach, 1 simulated tablet once a day. No other medication.  
Start in the morning on an empty stomach. 
6.4. 
Vaccine management 
Preservation conditions of the study vaccine: under 23°C, the vaccine was kept in a closed and 
dark place. 
 
The quality of the clinical trial vaccine, including the control tablet, should be checked regularly 
to meet the storage conditions. The test center would be reserved and managed by special personnel. 
 
The use of the study vaccine was the responsibility of the investigator, who should not transfer 
the study vaccine to any non-clinical trial participant. 
 
When distributing vaccine to subjects, researchers should fill in the registration form of vaccine 
distribution in a timely and accurate manner. 
After the end of the trial, the remaining vaccine would be collected for unified disposal. 
6.5. 
Combination of medicines and prohibited medicines 
 
Other anti-influenza medicines should not be used during the trial. 
 

 
 
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7. 
Clinical efficacy indexes and safety evaluation criteria 
7.1. 
Main efficacy indicators 
Immunogenicity of vaccines 
7.2. 
Other efficacy indicators 
 
Safety of vaccines 
 
Mortality 
 
7.2.1Immunogenicity of vaccines: 
 
To test the immunogenicity induced by oral vaccine 
 
Compared vaccine-induced immunogenicity with natural infection immunogenicity 
 
7.2.2 Safety of vaccines 
 
Observed whether mild to moderate symptoms such as fever and chills appeared after 
taking the vaccine; 
Observed whether there were severe symptoms such as coagulation dysfunction and 
allergy after taking the vaccine; 
Observed whether there were serious symptoms such as disability after taking the 
vaccine. 
 
7.2.3 Mortality 
 
Deaths were recorded for each group. 
 
7.3. 
Security indicators 
General data：physical examination  
 
Laboratory examination：Routine blood examination (including erythrocyte count, leukocyte count, 
hemoglobin, platelet count, proportion of monocytes, lymphocytes and 
neutrophils), urine routine (erythrocyte, leukocyte, urine protein), liver and kidney 
function (including ALT, AST, TBIL, BUN, Cr). 
 
Symptoms accompanying any discomfort were recorded by the investigator 
 
 
 
a) No: No symptoms

 
 
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b) Mild: Weak symptoms sometimes appeared 
c) Moderate: persistent but not noticeable after distraction 
d) Severe: Persisting no matter when 
7.4. 
Follow-up examination 
 
The first examination was performed on the day of study start (D0). Follow-up was 
conducted every 7 days (i.e. D7, D14, D21, and D28) for all clinical examinations, all 
indicators of the trial, and routine laboratory examinations. 
 
Side effect should be recorded during routine daily clinical examinations. If any side 
effect was found, follow up immediately and informed the researcher. 
 
7.4.1. 
When the test started 
 
 
Signed informed consent. 
Inclusion criteria and exclusion criteria. 
Medical history was collected and the status of medication was inquired. 
Routine physical examination. 
Routine laboratory tests: Routine blood examination (including erythrocyte count, leukocyte count, 
hemoglobin, platelet count, proportion of monocytes, lymphocytes and neutrophils), urine routine 
(erythrocyte, leukocyte, urine protein), liver and kidney function (including ALT, AST, TBIL, BUN, 
CR), pregnancy test (women of child-bearing age), and other laboratory examination. 
Combination medication was recorded. 
Assigned random numbers. 
8. 
Observation of side effect 
8.1. 
Definition 
 
Side effect: Any adverse medical event that occurred between the time patients signed the  
informed consent and were enrolled in the trial and the last follow-up, regardless of whether  
there was a causal relationship with the experimental vaccine, was considered as side effect. 
Serious side effect: Any side effect and hematological and/or other laboratory  
abnormalities that occur that lead to the use of targeted medical care. 
8.2. 
Criteria for determining the severity of side effect 
When completing the side effect table for CRF, the investigator would use mild, 
moderate, and severe to describe the intensity of the side effect. For unified standards, 
side effect intensity was classified as follows: 
 
Mild: Did not affect daily life;  
Moderate: Affected daily life; 
Severe: Severe affected on daily life. 
 
Be careful to distinguish between the severity and intensity of side effect. Severe was 
used to describe intensity and was not necessarily a serious side effect (SAE). For 
example, a headache might be severe in intensity but might not be classified as a serious 
side effect (SAE) unless it met SAE criteria. 

 
 
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8.3. 
Criteria for determining the relationship between side effect and the study vaccine 
The causal analysis of the relationship between all side effect (including serious side 
effect and general side effect) and the test vaccine was judged according to the five grades 
of almost certain, probable, possible, unlikely and unrelated. The first three side effects were 
defined as side effect to the vaccine. The following five aspects were causal analysis took 
into consideration: 
 
Was there a reasonable sequential relationship between the time of initiation and the  
occurrence of a suspected vaccine side effect (ADR) (appear when using vaccine). 
 
Did the suspected ADR conform to the type of ADR known to vaccines (consistent with the literature). 
 
Whether the suspected ADR could be explained by the effect of combination medication, previous  
medication, the subject's clinical status, or other therapies (other explanations). 
 
Did the suspected ADR disappear or decrease after a period of discontinuation of the vaccine. 
 
Whether suspicious ADR reappear (reuse) after re-exposure to the same vaccine. 
 
Causes analysis were carried out according to the following table, and the factors considered in the 
table were shown in the corresponding numerical table above. 
 
Factors 
1 
Appear 
when use 
vaccine 
2 
Same as 
reference 
3 
Other 
causes 
4 
Disappear 
when stop 
vaccine 
5 
Reappear 
when 
reuse 
Almost  
certain 
＋ 
＋ 
－ 
＋ 
＋ 
Probable 
＋ 
＋ 
－ 
＋ 
？ 
Possible 
＋ 
＋ 
± 
± 
？ 
Unlikely 
＋ 
－ 
± 
± 
？ 
Unrelated 
－ 
－ 
＋ 
－ 
－ 
 
The investigator should evaluate the possible association between side effect and the test 
vaccine and its combination using the following 5-level classification criteria: 
 
Almost  certain：Evidence of use of experimental vaccine; The chronological sequence 
of the occurrence of side effect and the administration of the experimental vaccine 
was reliable; The occurrence of side effect was more reasonably explained by the 
experimental vaccine than by other causes; The repeated vaccine test was positive; 
The pattern of side effect was consistent with what had been known about this or such 
vaccine. 
Probable：Evidence of use of the experimental vaccine; The chronological sequence of the 
occurrence of side effect and the administration of the experimental vaccine was reliable; side 
effect were more likely to be explained by the experimental vaccine than by other causes. 
Possible ：Evidence of use of experimental vaccine; The chronological sequence of the 
occurrence of side effect and the administration of the experimental vaccine was reliable; 
Side effect might be caused by the experimental vaccine or by other causes. 

 
 
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Unlikely：Evidence of use of the experimental vaccine; Side effect were more likely to be caused 
by other causes; Repeated vaccine tests were negative or equivocal. 
Unrelated：Patients did not take the experimental vaccine; Or the time sequence of the 
occurrence of side effect and the administration of the experimental vaccine could not 
be trusted; Or there were other significant causes that could lead to side effect. 
8.4. 
Determination of serious side effect 
 
side effect that met the following criteria (one or more) were classified as serious side effect
（SAE）： 
Die 
In danger of life, eg. in danger of immediate death 
Resulting in hospitalization 
Permanently or severely disabled 
Congenital malformation 
 
Medical events that had not yet resulted in death, life danger, or hospitalization should also 
be considered as SAE when appropriate medical judgment was made that they were likely to 
cause harm to the subject or that vaccines or surgical treatment was required to prevent the 
occurrence of these conditions. 
8.5. 
Record of serious side effect 
Any serious side effect in the course of a clinical trial must be reported to the sponsor, the 
principal study institution, the contract research organization, the ethics committee, National 
Medical Products Administration, and the ministry of health P.R.China within 24 hours (contact 
details were as follows). At the same time, the investigator must complete the Serious side effect 
Form, recording the time, severity, duration, action taken, and outcome of the serious side effect. 
 
9. 
Data administration 
9.1. 
Filing and handing over of case report form 
The case report form was completed by the investigator and must be completed by each selected case. 
After the completed case report form was reviewed by the clinical supervisor, the first copy was 
transferred to the data administrator for data entry and management. 
 

 
 
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9.2. 
Data entry and modification 
Data entry and management should be in the charge by the data administrator specially 
assigned by the group leader hospital. The data administrator prepared the data entry plan, 
carried on the data entry and the management. In order to ensure the accuracy of the data, two 
data entry personnel should independently conduct double entry and proofreading. 
 
For the questions in the case report form, the data manager would produce a question-
answer form and sent questions to the researcher through the clinical supervisor. The researcher 
should answer and return the questions as soon as possible. The data manager would modify, 
confirm and enter the data according to the answers of the researcher, and should send out the 
question-answer form again if necessary. 
9.3. 
Data locked 
After confirmation of the established database was correct under blind review, the main 
researchers, sponsors and statistical analysts would lock the data. The locked data file would 
not be changed. 
10. 
Statistic analysis 
10.1. Analysis data set 
Full analysis set：Refers to the collection of qualified cases and exclusions, but did not  
include exclusions. 
 
Per Protocol Set：Refers to the collection of cases that met the inclusion criteria and  
completed the treatment plan, i.e., met the trial protocol case, did not take 
the banned medicines, the case completed CRF and the prescribed 
contents. 
 
Security data set：Actual data on at least one treatment and documented safety indicators. 
10.2. Statistical analysis method 
10.2.1. Statistical analysis of population 
 
Intent-to-treat, ITT：The ITT population was defined as all patients who were randomized 
to double-blind treatment and had received at least one trial vaccine treatment with a 
corresponding efficacy evaluation. 
 
Per-Protocol, PP：PP population referred to the patients who had completed the vaccine 
treatment according to the protocol, and had completed all the evaluation contents 
without any significant deviation from the protocol. 
 
Safety Population：All patients who had used the study vaccine once and had undergone 
at least one safety assessment were randomized to be included in the study's safety 
population. Safety population was the main population for safety evaluation in this 
study. 
 
10.2.2. Statistical description 
 
Whether it conformed to normal distribution:  
Modified the statistical method or carried out data conversion in case of inconsistency; 
 

 
 
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With and without outliers: Conducted statistical and professional analysis to decide whether 
to choose or not; 
Missing value processing of primary efficacy index data: when a certain primary efficacy data 
of individual subjects was missing, the data would be collected from the statistics and 
professional standpoint. If the shedding case was missing, transferred with the previous 
measurement data; For missing data only one measurement index, all data was used to fill the 
gap. 
 
The proportion of shedding cases: If it exceeded 20%, it should be analyzed and explained; 
 
Descriptive statistical analysis: Indicated mean, standard deviation, maximum, minimum, median,  
confidence interval, and frequency (composition ratio), etc. 
 
10.2.3. Analytical method 
 
The experimental group was compared with the control group by superiority comparison. 
 
Measurement data: X2 test, t-test, paired t-test, rank sum test, paired rank sum test and non-
parameter test were used respectively statistical analysis to make statistical comparison and 
other methods. 
Descriptive statistical analysis of side effect was performed. 
 
10.2.4. Statistical expression 
 
The results of repeated measurements were presented in tabular form with statistical graphs in 
order to easy read. 
 
In general statistical tests, bilateral tests were used. If P was less than or equal to 0.05, it  
would be considered that the discriminant of the test was statistical significance. 
 
10.2.5. 
Statistical software 
 
The database was established by MS Access 
SPSS/SAS software was used for analysis. 
10.2.6. Contents of statistical analysis 
 
The statistical analysis plan was written by the statistician, finalized before the database 
lock, and consisted of a variety of tables.The main contents of the analysis included: 
 
Case distribution: different data sets in each group, case distribution in each center, comparison of 
total abscission rate, and detailed list of termination causes. 
 
Comparability analysis: compared demographic data with other base-value indicators to measure 
the comparability of the two groups. 
 
Effectiveness analysis: X2 test and t test were used for main indicators and global indicators. 
Since this study was a multi-center clinical trial, central effect on the efficacy indicators 
should be considered in the analysis. 
 
Analysis of the factors affecting the efficacy: such as age, sex, disease type, disease condition and so 
on, there were significant differences between the two groups before the use of vaccine, or 
if there were significant related factors that affected the vaccine effect, these factors should be 
used as co-factors when comparing the vaccine effect between the two groups for variables, 

 
 
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covariance analysis or Logistic regression analysis was required. The combination of medication 
and so on needed a detailed list. 
Safety analysis: firstly, according to the requirements of side effect correlation, the side effect 
and adverse reactions of the two groups were listed and described (including the number 
of cases of various side effects, the number of cases and the conversion rate of "normal to 
abnormal" or "abnormal intensification" of laboratory test indicators before and after the 
test), and the reasons and explanations were listed. 
11. 
Quality control and assurance 
11.1. Quality control 
During the study, clinical inspectors will be commissioned by the sponsor to visit each study 
center regularly to ensure that all the contents of the study plan were strictly followed and the 
study data were filled in correctly. 
 
The whole clinical trial process strictly followed the provisions of the national GCP (Good 
Clinical Practice) and conformed to the ethical principles. Participants might strictly abide by 
the standard operating procedures of clinical trial study. 
 
Participants in clinical trials should be relatively fixed, and participants might undergo 
uniform training and had uniform recording methods and judgment criteria. 
 
The researcher should fill in the CRF truthfully, carefully and use pen or carbon pen. 
 
All observations and findings in clinical trials should be verified to ensure the reliability of 
the data and to ensure that the conclusions of clinical trials were derived from the original data. 
There were corresponding data management measures in clinical trial and data processing 
stage. 
 
Medical records and medical record forms were regarded as the original records and 
generally could not be changed. When making any corrections, the original record should not 
be altered. Additional statement could be used to explain the reason, and then signed and dated 
by the physician participating in the clinical trial. 
 
The researchers should take active measures (notice further consultation, follow up) to  
control the shedding rate within 20%. 
 
Subjects would record their vaccination status on a diary card. 
 
Each center should submit its normal range and abnormal judgment criteria to the 
responsible unit of clinical trial and the clinical statistical unit to ensure the accuracy, reliability 
and abnormal judgment criteria of laboratory examination. 
 
All laboratory data should be recorded in the original medical record and the results of the 
tests should be glued to the original medical record. Laboratory data within the normal range 
should also be recorded in the CRF, and data that were significantly higher or outside the 
clinically acceptable range should be verified by the physician participating in the clinical trial 
to make the necessary statements. 
 
 
Administration of vaccines: 
Preservation conditions of experimental vaccine: under 23°C, sealed in a dark place. 

 
 
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Periodic review of clinical trial vaccines, including the quality of control tablets; 
Conformed to storage conditions. The test center would be reserved and managed by 
special personnel. 
The investigator was responsibility to the used test vaccine, who should not transfer the 
test vaccine to any non-clinical trial participants in the test. 
 
When distributing vaccine to subjects, researchers should fill in the registration form of vaccine 
distribution in a timely and accurate manner. 
 
After the end of the trial, the remaining vaccine would be collected for unified  
disposal. 
11.2. Quality guarantee 
11.2.1. Establish a multi-center coordination committee 
 
The group leader should be responsible for the organization of the clinical trial center and 
the sponsor to form the clinical coordination committee. The main task of the coordination 
committee was to coordinate the implementation of the whole clinical trial and to study and solve 
the problems arising from the clinical trial. The sponsor was responsible for maintaining contact 
with the National Medical Products Administration. 
 
11.2.2. Enact standard operating procedures for clinical inspectors 
 
Supervisors appointed by the sponsor would monitor the whole process of the clinical trial. 
This included ensuring that subjects' rights and interests were protected and clinical trials were 
conducted in strict process in accordance with the requirements of clinical trial protocols and 
relevant laws and regulations. 
12. 
Medical ethics and related legal issues 
12.1. Guidelines for clinical trial on vaccines 
The clinical trial would be conducted in accordance with the Helsinki Declaration and the 
clinical quality management practice (GCP) for vaccines. Before the start of the study, the study 
plan had been approved by the ethics committee of the hospital of the study leader unit. At the 
same time, the ethics committee would play a supervisory role throughout the trial process. 
12.2. Investigator responsibility 
The investigator might ensure that all participants in the study had a detailed 
understanding of the study protocol, study procedures and evaluation methods, as well as their 
responsibilities and roles. 
12.3. Study subjects and informed consent 
Prior to the inclusion of patients in the study, the investigator might provide a complete and 
comprehensive description of the purpose, procedures, and possible risks of the study to the patient or 
his or her designated representative. Patients should be made aware of their right to withdraw from the 
study at any time. Each patient might be given a written patient informed consent form prior to study. 
The study physician was responsible for obtaining signed informed consent form from each patient 
before the study and sending one of the signed informed consent forms to the subjects; The other was 
kept in the study archives. Any changes to the subject's informed consent might be documented and 
submitted to the IRB for approval. A copy of the new Informed Consent Form would be kept in the 
"Subject Informed Consent Form" file. Suggestions for modification were attached. The above changes 
could only be implemented if they were re-approved by the ERC, complied with the health regulations, 

 
 
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and signed by the participating institutions. Each subject or independent witness affected by the 
modification and any person involved in the informed consent discussion might sign and date two 
updated informed consent forms. He or she would be given a new form of informed consent. If the 
subject had a health care provider and agrees to notify the health care provider, it was recommended 
that the investigator explained the subject and the study to the subject's health care provider. 
12.4. Implementation and modification of the study plan 
Prior to the commencement of the study, the study protocol, informed consent, and other 
relevant materials would be submitted to an independent institutional review board (IRB), 
together with a table of contents or list. Clinical trials could commence only after the applicant 
had received all the ethical and legal documents necessary. These documents included a list of 
IRB members and their occupations and qualifications. If the IRB did not disclose the name of 
the committee, they might issue a statement assuring that the composition of the committee met 
the GCP standards. The formal IRB approval should mention the study topic, study number, 
study location, and other relevant documents. Documents might refer to the date of the decision 
and be signed by the members of the Committee. 
 
Neither the investigator nor the organizer could change the protocol without prior written 
notice. Once the study plan was launched, any problems were found in the implementation 
process could be modified and improved under the jointly negotiated by the sponsor and the 
researcher. A document should be written and signed by the main researcher and the sponsor 
before implementation. At the same time, a report should be submitted to the ethics committee 
and the documents should be filed. 
12.5. Termination of study 
The sponsor or investigator had the right to decide completion time of the study. After 
communication between both sides noticed IRB as required by law. 
 
Study materials (including completed, partially completed and blank case sheets, study 
vaccines, etc.) might be returned to the sponsor when the study might end early. 
12.6. Relevant legal liability 
The applicant should be legally liable for the losses caused by the participants in compliance with the plan,  
law and professional standards. 
13. 
Summary of test report 
The data management and statistical analysis of this clinical trial were completed by the 
_____Statistics Teaching and Research Section. After the statistical analysis report was 
completed, the general statistical analysis report of this study and the statistical report of each 
analysis center would be written. 
 
According to the statistical report, each experimental unit should write the summary of the 
sub-center, and the study leader should complete the summary report of the clinical trial and 
affixed the official seal. 
 
14. 
Data save 
In order to ensure the evaluation and supervision of the National Medical Products 
Administration and the sponsor, under normal circumstances, all study data records were stored 

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by the researchers after the end of the study, and the retention period was 5 years. 
 If the researcher failed to meet the preservation requirements, alternative arrangements 
might be made with the consent of the sponsor. 
The investigator might obtain the Sponsor's written consent before destroying the records. 
All the data of this clinical trial belonged to the sponsor. Except for the requirements of the National 
Medical Products Administration, the investigator should not provide them to any third party in any way 
without the written consent of the sponsor. 
15.
Publication and use of study results
DreamTec Research Limited (DTRL) was the owner of the data and results obtained in
this study. If the researcher published the relevant content in journals or academic conferences, 
he/she might obtain the consent of the DTRL in advance. 
16.
Test plan
Date 
Milestone 
（1st day） 
Completed the formulation of the trial 
protocol through ethics 
Each center began to enroll the first 
case in the screening period 
All 
the 
centers 
completed 
the 
enrollment of all the patients during 
the trial period 
All centers completed the follow-up 
work of all cases 
（90th days） 
Completed data entry and statistics 
work 
Completed the test summary report 
Ended of all trials 

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21 
Research Team / Medical Consultants 
Name 
Position / Organization 
Relevant 
Qualifications 
/ 
Working 
Experience for the Project 
Dr. Eric Tung Po Sze (Co-
I) 
Associate 
Professor, 
Hong 
Kong 
Metropolitan 
University 
Ph.D. Chemistry (CUHK) 
M.Sc. Applied Toxicology (University of
Surrey, United Kingdom)
Dr. 
Keith 
Wai 
Yeung 
Kwong (Co-PI) 
Chief Scientific Officer 
and 
Executive 
Director, 
DreamTec 
Research Limited 
B.Sc. 
in 
Biochemistry 
(University 
of
Washington, USA)
Ph.D. in Biochemistry (Molecular Medicine)
(HKUST)
Dr. Liu Ying (Research 
Team Members) 
Chief Medical Officer, 
DreamTec 
Research 
Limited 
B.Sc. in Clinical Medicine (LiaoNing Medical
College, China)
Ph.D. in Biophysics (Peking University,
China)
Post-doctoral Fellow in Division Biomedical
Engineering (HKUST)
Senior Research Associate in Department of
Biomedical Sciences (City University of Hong
Kong)
Technical 
Manager 
of 
Vanway
Pharmaceutical Holdings Limited
Medical 
Consultant, 
Health 
& 
Beauty
International Holdings Limited
Functional 
& 
Integrative 
Medicine, 
LifeHealth 
Group 
Limited 
M.B. B.S. (London)
MRCP (United Kingdom)
DRCOG
Dr. 
Stephen 
Chan 
(Medical Consulant) 
Chief Medical Officer, 
LifeHealth 
Group 
Limited 
M.B. B.S. (NUS)
MMed Fam. Med. (NUS)
LMCHK
Dr. See Yunn Ho (Medical 
Consultant) 

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Informed Consent form 
TITLE OF THE STUDY 
The Development of a COVID19 Oral Vaccine Consisting of Bacillus Subtilis Spores 
INTRODUCTORY SENTENCE 
You are invited to participate in a research study: 
The Development of a COVID19 Oral Vaccine Consisting of Bacillus Subtilis Spores.
Bacillus subtilis RBD Spores is a health supplement that enhances your immunity to generate and boost 
neutralizing antibody to prevent the virus from attacking the human body. Postbiotics formula contains “Bacillus 
subtilis coating with RBD of spike protein in SARS-COV2”, It induces a series of immune responses in human 
body to generate significant immune affection therefore provide protection against the viruses. 
Features 
1. Improving immunity
2. Boost antibody
3. Safe and minimum side effects
PURPOSE OF THE STUDY 
To test the antibody level during the period of consuming the Bacillus subtilis RBD Spores 
PROCEDURES 
-
Participants who understand the plan of health food and trial, confirmed meet the guideline and completed
consultation with program staff.
-
During the trial, the participants must follow the consumption guideline.
-
The trial period is 44 days, which included 3 Box (9capsules), laboratory tests and a questionnaire. The trial and
questionnaire content are as follows:
1.
It is recommended to stop taking any immune enhancement health products during the trial
2.
Total 9 capsule, consumption 1capsule on day 1, day 2, day 3, day 14, day 15, day 16, day 28, day 29 and
day 30 respectively
3.
Please fill in the Form during the trial
POTENTIAL RISKS/STRESS/PAIN/DISCOMFORTS/OTHER FACTORS AND THEIR MINIMIZATION 
-
If you are taking other medicines or similar function drugs at the same time, please consult a doctor before trial
-
If you have any drugs allergy, please consult a doctor before trial
-
This trial may cause discomfort such as: diarrhea
-
If you have any discomfort caused by your own related diseases or physical conditions during the test, please stop
taking this product and consult a doctor immediately
-
Blood sample is required before this trial
-
People who have not been vaccinated. If you accept to start the trial (44 days), you will not be vaccinated during
this period. If you vaccinate, the participant at their own risks. Due to the termination of the test, the participant
shall be responsible for any cost for this trial
-
This plan is self-contained, program in charge shall not be held responsible for any accidents and/or personal
injuries howsoever caused to the participant and the participant at their own risks
POTENTIAL BENEFITS 
-
Enhances participants immunity to generate and boost neutralizing antibody to prevent the virus from attacking
the human body.

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PARTICIPATION AND WITHDRAWAL 
-
Your participation is entirely voluntary and that you can choose to withdraw from the study at any time you want
without any penalty or negative consequences.
CONFIDENTIALITY 
-
The test report will be kept by program center
QUESTIONS AND CONCERNS 
If you have any questions or concerns about the research study, please feel free to contact Dr. Sunny Wong of 
Doctor Think Tank Academy at 6846-8772. If you have questions about your rights as a participant of this research 
study, please contact the Research Ethics Committee of Doctor Think Tank Academy at admin@dttac.com. 
<Please complete by a participant> 
Participant No. (will be provided by the program staff) 
_______________ 
Hypothesis (will be provided by the program staff) 
______________ 
Consent 
I have read and understood this trial, and the product developer has also provided me with 
detailed explanations and answered all my queries. I have already known the purpose and risk 
of this trial and voluntarily consent to participate in the trial described above. I have 
been given a copy of this consent form. 
Paticipant      :_____________________________ 
Paticipant Signature :__________________________ 
Phone no.      : _____________________________ 
Date     
: _____________________________