A Single-center, Open, Single-arm Clinical Study on the Application of Ursodeoxycholic Acid for the Prevention of SARS-CoV-2 Infections
Phase: PHASE1
Conditions: COVID-19
Sponsor: Institute of Hematology & Blood Diseases Hospital, China

 
A single-center, open, single-arm clinical study on the 
application of ursodeoxycholic acid for the prevention of 
novel coronavirus infections 
 
 
 
PI 
Lei Zhang 
Research Unit： 
Blood Diseases Hospital, Chinese Academy of Medical 
Sciences 
Version number： 
V2.0 
Version Date： 
2023.3.6 
ID 
2022UDCA-COVID-19 
 
 
 
 
 
 
 
 
 
 
 
 
 
Confidentiality Statement 
This document is confidential information of the Hospital of Hematology, Chinese Academy 
of Medical Sciences (Institute of Hematology, Chinese Academy of Medical Sciences) and is 
intended for the purpose of this clinical study only. It may not be disclosed to anyone other than the 
participants in the study and members of the Institutional Review Board. This information may not 
be used for any purpose other than the evaluation or conduct of the clinical study without the prior 
written consent of the Hospital of Hematology, Chinese Academy of Medical Sciences (Institute of 
Hematology, Chinese Academy of Medical Sciences). 
 

 
Confidential 
Researcher Statement 
I have familiarized myself with the protocol of this study and confirm that it 
contains the necessary elements for the implementation of the study and that the 
responsibilities related to this study protocol are clear. I agree to perform the relevant 
duties in accordance with Chinese laws and regulations, the Declaration of Helsinki, 
the Code of Quality Management for Drug Clinical Trials, and this study protocol, and 
to carry out the procedures specified in this study only after the Ethics Committee has 
approved the protocol and the subjects have given their informed consent. If 
modifications to the protocol are required, the modified protocol will be approved by 
the Ethics Committee before implementation, unless measures are necessary to protect 
the safety, rights, and interests of the subjects. I understand and comply with the 
requirements for the maintenance of original data. 
 
 
 
 
 
 
 
 
Principal Investigator 
 
Date 
 
 
 
 

 
 
Contents 
1 
ABSTRACT ................................................................................................................................................................ 1 
2 
RESEARCH BACKGROUND .................................................................................................................................. 1 
3 
RESEARCH OBJECTIVES ...................................................................................................................................... 1 
3.1 
PRIMARY RESEARCH OBJECTIVES .......................................................................................................................... 1 
3.2 
SECONDARY RESEARCH OBJECTIVES ..................................................................................................................... 1 
4 
STUDY DESIGN ........................................................................................................................................................ 2 
5 
STUDY POPULATION ............................................................................................................................................. 2 
5.1 
INCLUSION CRITERIA ............................................................................................................................................. 2 
5.2 
SHEDDING AND EXIT CRITERIA .............................................................................................................................. 3 
5.3 
REJECTION CRITERIA ............................................................................................................................................ 3 
6 
DRUG DELIVERY MEASURES ............................................................................................................................. 3 
6.1 DRUG DELIVERY METHOD/PROTOCOL .......................................................................................................................... 3 
7 
VISITING ARRANGEMENTS................................................................................................................................. 3 
7.1 SCREENING PERIOD ..................................................................................................................................................... 3 
7.2 DOSING PERIOD (4 WEEKS) .......................................................................................................................................... 4 
7.3 FOLLOW-UP PERIOD (8 WEEKS) .................................................................................................................................... 4 
8 
RESEARCH INDICATORS ...................................................................................................................................... 7 
8.1 PRIMARY RESEARCH INDICATORS................................................................................................................................. 7 
8.2 SECONDARY RESEARCH INDICATORS ............................................................................................................................ 7 
8.3 SAFETY INDICATORS .................................................................................................................................................... 7 
9 
ADVERSE EVENTS AND SERIOUS ADVERSE EVENTS.................................................................................. 7 
9.1 ADVERSE EVENTS ....................................................................................................................................................... 7 
9.2 SERIOUS ADVERSE EVENTS ......................................................................................................................................... 9 
10 
DATA MANAGEMENT .......................................................................................................................................... 10 
11 
STATISTICAL METHODS ..................................................................................................................................... 10 
11.1 SAMPLE SIZE ............................................................................................................................................................ 10 
11.2 STATISTICAL ANALYSIS SET ...................................................................................................................................... 11 
11.3 STATISTICAL ANALYSIS METHODS ............................................................................................................................ 11 
12 
ETHICS REVIEW ................................................................................................................................................... 12 
12.1 ETHICAL REQUIREMENTS ......................................................................................................................................... 12 

 
 
12.2 INFORMED CONSENT ............................................................................................................................................... 12 
12.3 CONFIDENTIALITY OF INFORMATION ........................................................................................................................ 13 
13RESEARCH MANAGEMENT ................................................................................................................................... 13 
13.1 TRAINING ................................................................................................................................................................ 13 
13.2 QUALITY CONTROL AND ASSURANCE ....................................................................................................................... 14 
14 CONFIDENTIALITY AND PUBLICATION OF RESEARCH RESULTS ........................................................... 14 
15 REFERENCES ............................................................................................................................................................ 14 
16 ANNEXES .................................................................................................................................................................... 15 
 

 
 
1 
1 
ABSTRACT 
Title： 
A single-center, open, single-arm clinical study on the application of ursodeoxycholic acid for the 
prevention of novel coronavirus infections 
Planned research time： 
Expected to enroll healthy volunteers for 3 weeks and follow up for 8 weeks 
Purpose of the study： 
1) Primary research objectives 
Evaluation of the efficacy and safety of ursodeoxycholic acid in blocking the transmission of 
infectious novel coronaviruses in a population of medical workers 
2) Secondary Research Objectives 
Exploring the evaluation of conversion to severe disease in people infected with novel 
coronavirus 
Exploring the evaluation of the persistence and reduction of symptoms in people infected 
with novel coronavirus 
Exploring the evaluation of antibody profiles in healthy volunteer populations 
Observing the safety of ursodeoxycholic acid drugs 
Evaluating the changes of S protein-specific T/B cell immunity before and after BF.7 
infection after immunization with Wuhan Strain vaccine. 
 
Study population： 
Healthy volunteers for this study were from medical workers (doctors, nurses, medical 
technicians, administrative logistics, medical students) who were not infected with novel 
coronavirus 
Planned enrollment (sample size)： 
The study is expected to enroll 130 healthy volunteers 
Study Design Overview： 
This study is a single-center, open-ended, single-arm clinical study 
Inclusion Criteria： 
Healthy volunteers suitable for enrollment in this study must meet all of the following criteria： 
1) Age 18-65 years old 
2) No restriction on gender 
3) Medical-related personnel (working hours >20 hours per week) serving during the COVID-
19 outbreak, including physicians, nurses, medical technicians, administrators, and medical 
students 
4) COVID-19 nucleic acid test negative within 48 hours 
5) Sign the informed consent form 
Exclusion Criteria： 
Anyone who meets any of the following criteria shall not be selected for this study： 

 
 
2 
1) Exhibit COVID-19 symptoms, including fever, muscle pain, headache, cough, sore throat, 
and loss of smell and taste 
2) Previous infection with novel coronavirus within 6 months 
3) Previous (past 30 days), current or planned (during the study period) use of 
immunomodulatory-related drugs 
4) Those with allergy or intolerance to ursodeoxycholic acid, gallbladder or bile duct disease, 
severe liver failure, or liver function impairment 
5) Pregnancy and lactation 
6) Use of drugs with which ursodeoxycholic acid is contraindicated 
7) Other reported health conditions that make participation in the study not in the best interest 
of the individual 
 
Shedding and exit criteria  
1) Healthy volunteers request to withdraw from the study on their own 
2) Female healthy volunteers planning to become pregnant or pregnant during the trial 
3) Missed visits due to various reasons 
4) In the judgment of the investigator, treatment should be discontinued from this protocol 
Rejection Criteria： 
1) Poor compliance with the requirements of the study and failure to follow the study plan 
2) Those who received prophylactic drugs other than this drug during the trial 
Research drug delivery protocols： 
Drug name (specification formulation): ursodeoxycholic acid capsules. 
Mode of administration/regimen: 250 mg/capsule, 2 capsules per day, for 4 weeks. 
Duration of follow-up visits： 
Dosing period： 
Healthy volunteers took ursodeoxycholic acid capsules daily for 4 weeks from the date of 
enrollment 
Follow-up period： 
Healthy volunteers will be followed up once a week for a total of 8 weeks in case of new 
coronavirus infection. 
Research Indicators： 
1) Primary research indicators 
Prevalence of novel coronavirus infection in healthy volunteers receiving 4 weeks of 
continuous ursodeoxycholic acid plus conventional protection during the study period. 
2) Secondary Research Metrics 
1) The proportion of people infected with novel coronavirus who turned severe. 
2) the number of days that symptoms of a novel coronavirus infection such as fever, sore 
throat, headache, and muscle aches persisted in the population infected with novel coronavirus, 
and the number of days that the nucleic acid/antigen turned negative. 
3) The rate of positive serological antibodies in the population of healthy volunteers. 
4) The incidence of adverse events. 

 
 
3 
5) The difference of T/B lymphocyte specific response to Wuhan Strain and BF.7 Strain in 
different volunteers. 
Safety evaluation： 
Observe and record any adverse events that occur in healthy volunteers during the clinical 
study, including abnormal clinical symptoms and vital signs, abnormalities in laboratory tests, 
record the characteristics of clinical manifestations, severity, time of occurrence, duration, 
management and prognosis, and determine their correlation with the trial drug/treatment regimen, 
and evaluate safety according to NCI CTCAE version 5.0 standards 
Statistical Methods： 
SAS 9.4 software was used for statistical analysis of the data in this study, with continuous 
variables described by mean ± stan dard deviation (Mean ± SD) or median, maximum, minimum, 
and interquartile range (Median, Max, Min, IQR) and categorical variables described by rate or 
percentage. Comparisons of measurement data were performed by t-test/ANOVA or 
nonparametric test; comparisons of count data were performed by χ2 test or Fisher's exact 
probability test. If not otherwise specified, all tests were performed as two-sided tests with a test 
level of α=0.05. 

 
1 
2 
Research Background 
Researchers at the University of Cambridge, UK, and other institutions have found that 
ursodeoxycholic acid (UDCA) is able to turn off the ACE2 receptor, thereby blocking viral 
entry into human cells, and since the SARS-CoV from 2003 to the SARS-CoV-2 spectrum in 
2019 utilizes ACE2 as the primary invasion receptor, even if subsequent SARS-CoV-2s 
continue to show new small variants, the drug maintains its effect. The study was published in 
Nature on December 5, 2022, entitled "FXR inhibition may protect from SARS-CoV-2 
infection by reducing ACE2" [1]. Ursodeoxycholic acid (UDCA) was found to downregulate 
angiotensin-converting enzyme 2 (ACE2) expression by inhibiting the transcription factor FXR, 
thereby shutting down the entry of neocoronavirus into human cells. Since the drug acts on the 
host cell and not on the virus itself, it may be able to prevent current neo-coronavirus infections 
and should also be able to prevent future neo-coronavirus new mutants, as well as infections by 
other coronaviruses that utilize ACE2. Currently, China is in a period of high prevalence of 
neo-coronavirus infections, and there is an urgent need for clinical studies on prophylactic drugs 
 
3 
Research objectives 
The purpose of this study is to evaluate whether ursodeoxycholic acid is an effective 
prophylactic agent against novel coronavirus (SARS-CoV-2) infection in medical personnel 
3.1 Primary research objectives 
Evaluation of the effectiveness and safety of ursodeoxycholic acid in blocking the transmission 
of infectious novel coronaviruses in adult populations 
3.2 Secondary research objectives 
1) To explore the evaluation of the transition to severe disease in the population infected 
with novel coronavirus; 
2) To explore the evaluation of the persistence and reduction of symptoms in people 
infected with novel coronavirus; 
3) To explore the evaluation of antibodies in a population of healthy volunteers; 
4) To observe the safety of ursodeoxycholic acid drugs; 
5) To evaluating the changes of S protein-specific T/B cell immunity before and after 
BF.7 infection after immunization with Wuhan Strain vaccine. 

 
2 
4 
Study design 
This is a single-center, open-ended, single-arm clinical study to evaluate the effectiveness 
of daily ursodeoxycholic acid plus routine protective measures for the prevention of novel 
coronavirus infection. The target sample size is 130 medical workers and each healthy volunteer 
will be followed weekly for nucleic acid/antigen testing or disease progression over an 8-week 
period for new crown infections. 
 
5 
Study population 
Healthy volunteers for this study were drawn from medical workers who were not infected 
with novel coronavirus 
5.1 Inclusion criteria 
Healthy volunteers suitable for enrollment in this study must meet all of the following 
criteria： 
1) Age 18 - 65 years old. 
2) No restriction on gender. 
3) Medical-related personnel (working hours >20 hours per week) serving during the 
COVID-19 outbreak, including physicians, nurses, medical technicians, administrators, 
and medical students 
4) COVID-19 nucleic acid test negative within 48 hours 
5) Sign the informed consent form 
5.2 Exclusion Criteria 
Anyone who meets any of the following criteria shall not be selected for this studies： 
1) Exhibit COVID-19 symptoms, including fever, muscle pain, headache, cough, sore 
throat, and loss of smell and taste 
2) Previous infection with novel coronavirus within 6 months 
3) Previous (past 30 days), current or planned (during the study period) use of 
immunomodulatory-related drugs 
4) Those with allergy or intolerance to ursodeoxycholic acid, gallbladder or bile duct 
disease, severe liver failure, or liver function impairment 
5) Pregnancy and lactation 
6) Use of drugs with which ursodeoxycholic acid is contraindicated 

 
3 
7) Other reported health conditions that make participation in the study not in the best 
interest of the individual 
5.3 Off study criteria 
Volunteers withdrew from the study for any of the following reasons. 
1) A healthy volunteer's own request to withdraw from the study. 
2) planned pregnancy or pregnancy during the trial in female healthy volunteers. 
3) Loss of visit for various reasons. 
4) The investigator's judgment should discontinue treatment in this protocol. 
Healthy volunteers may withdraw their informed consent if they do not wish to continue to 
participate in the study. If a healthy volunteer decides to withdraw from the study, the 
investigator must be notified in a timely manner. The reason for withdrawal from the study 
should be documented, including the date and reason for withdrawal from the study. 
 
5.4 Rejection Criteria 
Volunteers were required to be excluded from the study for the following conditions 
1) Those who are poorly compliant with the requirements of the study and fail to follow the 
study plan. 
2) Those who receive prophylactic medications other than this drug during the trial. 
 
6 
Drug delivery measures 
Drug Name (Specification Formulation). 
Ursodeoxycholic acid capsule, 250mg/capsule. 
6.1 Drug delivery method/protocol 
Take two capsules orally with a meal, once daily 
 
7 
Visiting Arrangements 
7.1 Screening Period 
• 
Baseline Collection 
1) Basic demographic information such as age and sex. 
2) Type of neo-coronavirus vaccination and number of vaccinations received. 
3) novel coronavirus infection status of family members/personnel living with them. 
4) Personnel positions: e.g., physicians, nurses, medical technology, administrative 
logistics, medical students, etc. 

 
4 
5) Working department. 
6) underlying disease status: e.g. hypertension, diabetes, cardiovascular disease, etc. 
7) Combined medication use and drug allergy history. 
• 
Routine Inspection： 
1) Vital signs (heart rate, respiration, body temperature, etc. 
• 
Retention of serum samples： 
1) 5mL of blood was collected from healthy volunteers, of which 2mL of whole blood 
was directly frozen and the rest was extracted from the serum for freezing and storage. 
2) Biological sample processing and storage. 
Blood samples will be collected according to the follow-up plan, and the processing and storage 
will be carried out by the Biosample Bank of the Hospital of Blood Diseases, Chinese Academy 
of Medical Sciences. It will be managed in strict accordance with biosafety law, regulations on 
human genetic resources management and other relevant regulations. 
7.2 Dosing period (4 weeks) 
1) weekly nucleic acid testing (in lieu of nucleic acid results if positive for self-test 
antigens) 
2) Weekly telephone follow-up for adverse events. 
3) 5mL of blood is collected from healthy volunteers at the end of dosing or in case of 
infection during the period, of which 2mL of whole blood is directly frozen and the rest is 
extracted from the serum for freezing. 
7.3 Follow-up period (8 weeks) 
1) If a healthy volunteer develops novel coronavirus symptoms during the follow-up 
period, the volunteer's symptoms will be scored and recorded according to Table 1. 
Table 1: New crown symptom assessment form 
Symptoms 
Rating 
Slight =1 
Medium =2 
Serious =3 
Score 
Abdominal 
pain 
 
 
 
 
fatigue 
 
 
 
 
Cough 
 
 
 
 
Shortness of 
breath 
 
 
 
 
Diarrhea 
 
 
 
 
Muscle pain 
 
 
 
 
Headaches 
 
 
 
 
Taste and 
smell disorder 
 
 
 
 

 
5 
Other 
 
 
 
 
Total Score 
 
 
 
 
 
 
1) Weekly telephone follow-up of AE status of healthy volunteers. 
2) Detection of antibodies before and after drug administration. 
3) Weekly monitoring of nucleic acid/antigen status of new crowns in healthy volunteers 
during the study period, and testing for nucleic acid at any time during the study period 
when volunteers develop symptoms of novel coronavirus (in case of positive self-test 
antigen, this can be substituted for nucleic acid results)./ 
4) Blood will be collected for freezing and storage in case of infection and out of the 
group. 
 
Table 2: Table of observation items during the visit 
 
Baseline 
period 
Detection 
frequency 
In case of 
infection 
Demographic 
Information 
 
 
 
Nucleic acid testing 
 
Weekly 
 
Vital signs 
 
 
 
AE Telephone 
follow-up AE 
 
Weekly 
 
Blood collection 
for freezing and 
storage 
 
Week 4 and 8 
 
 
 

 
6 
Figure 1: Research flow chart 
 
 
Figure 2: Flow chart of the visit 

 
7 
8 
Research indicators 
8.1 Primary research indicators 
Prevalence of novel coronavirus infection in healthy volunteers receiving 4 weeks of 
continuous ursodeoxycholic acid plus conventional protection during the study period, the 
proportion of healthy volunteers infected with novel coronavirus during the study period as a 
percentage of the study population 
8.2 Secondary research indicators 
1) The proportion of people infected with novel coronavirus who turned severe. 
2) the number of days that novel coronavirus symptoms such as fever, sore throat, 
headache, and muscle aches persisted in the population infected with neo-coronary, and the 
number of days that the nucleic acid/antigen turned negative. 
3) The rate of positive serological antibodies in the population of healthy volunteers. 
4) The incidence of adverse events. 
5) The difference of T/B lymphocyte specific response to Wuhan Strain and BF.7 Strain 
in different volunteers. 
8.3 Safety indicators 
Observation and recording of various adverse events during administration of 
ursodeoxycholic acid in healthy volunteers 
 
9 
Adverse events and serious adverse events 
All adverse events, including abnormal clinical symptoms and vital signs, and 
abnormalities in laboratory tests from the date of signing the informed consent to the last visit 
were recorded, and the characteristics of clinical manifestations, severity, time of occurrence, 
duration, management and prognosis were recorded, and their correlation with the trial 
drug/therapeutic regimen was determined. 
Adverse events were graded and recorded with reference to NCI CTCAE 5.0 
9.1 Adverse Events 
9.1.1 Definition of Adverse Events 
Adverse event:  An adverse medical event that occurs after a healthy volunteer 
participates in a clinical study, not necessarily related to the study treatment, and includes any 

 
8 
new event that occurs or worsens in severity and frequency compared to the baseline situation, 
including abnormal results of laboratory tests, physical examinations, etc. Adverse events 
include Severe adverse event (SAE), Adverse event (AE), and abnormal laboratory results. 
9.1.2 Adverse event collection and recording 
Adverse events from the date of signed informed consent to the last visit were recorded, 
and the investigator applied concise medical terminology to report all possible treatment-related 
adverse events. 
9.1.3 Criteria for determining the severity of adverse events 
Use the National Cancer Institute Common Terminology Criteria for Adverse Events 
(CTCAE) version 5.0 to document the severity of adverse events on a scale of 1-5 
 
Grading 
Criteria 
1 
Mild; asymptomatic or mild; seen clinically or diagnostically only; no 
treatment required 
2 
Moderate; requires lesser, localized or non-invasive treatment; age-
appropriate limitation of instrumental activities of daily living* 
3 
Severe or medically significant but not immediately life-threatening; 
resulting in hospitalization or prolonged hospitalization; disabling; limiting 
personal activities of daily living △ 
4 
Life-threatening; requires urgent treatment 
5 
AE-related deaths 
* Instrumental activities of daily living: refers to cooking, buying clothes, using the 
telephone, managing money, etc. 
△Personal activities of daily living: refers to bathing, dressing and undressing, eating, 
toileting, taking medication, etc., and not bedridden.。 
Note:the distinction between severity and intensity of the adverse event. Severity is used 
to describe intensity and is not necessarily an SAE. e.g. a headache may be severe in 
intensity but cannot be included in an SAE unless it meets SAE criteria. 
9.1.4 Criteria for judging the relationship between adverse events and  
study drug therapy 
The investigators analyzed the relevance of adverse events to the study drug treatment 
according to the principle of determining the relevance of adverse events to the study drug 

 
9 
treatment, which was divided into 5 categories, namely "definitely related, probably related, 
probably unrelated, definitely unrelated, and undetermined", and "definitely related, probably 
related, probably unrelated, and The adverse reactions were classified as "definitely related, 
probably related, probably unrelated and undetermined". The investigator must document the 
adverse reactions using medical terminology. 
 
Is it reasonable 
Time relationship 
Is it consistent with known 
Type of adverse reaction 
Whether there are other reasons 
forThe possibility of adverse 
events 
Definitely 
related 
Yes 
Yes 
Yes 
Possibly 
related 
Yes 
Yes / No 
Uncertain 
Possibly 
unrelated 
No 
No 
Uncertain 
Definitely 
not related 
No 
No 
Yes 
Unable to 
judge 
Required information for evaluation is not available 
 
9.2 Serious Adverse Events 
9.2.1 Definition of serious adverse events 
1. serious adverse event can be defined as any of the following. 
1) Deadly or life-threatening. 
2) requiring hospitalization or extended hospitalization. 
3) Resulting in significant or lasting disability or functional impairment. 
4) causing a congenital anomaly or birth defect 
5) medically significant and serious illness that endangers the patient's health and requires 
medical intervention or surgical intervention to prevent any of the above outcomes. 
2. The following admissions are often not considered serious adverse events. 
1) Discharge or withdrawal of the patient for progressive exacerbation of the disease 
requiring hospitalization or extended hospitalization. 
2) Hospitalization for routine treatment or observation for indications involved in the trial 
that are not accompanied by a worsening of the disease; 

 
10 
3) the patient's hospitalization was planned prior to enrollment in this trial and no 
exacerbation of symptoms that existed prior to the trial was observed 
4) Treatment is received only in an emergency, outpatient setting that does not meet any 
of the above definitions of a serious adverse event and does not require hospitalization. 
5) The pregnancy itself is not a serious adverse event, but should be reported on a serious 
adverse event form or pregnancy record form and should be followed up as well as the 
outcome documented, including spontaneous abortion or voluntary termination of pregnancy, 
details of the birth of the infant, and the presence of congenital anomalies and birth defects. 
9.2.2 Reporting of serious adverse events 
If a Grade 5 serious adverse event occurs in a healthy volunteer during the course of the 
study (from the date of signing the informed consent to the last visit) and may be related to 
the study drug treatment, the investigator must complete a serious adverse event report form 
and report it to the ethics committee of the study center where the event occurred within 24 
hours of learning of the event, and the investigator must sign and date the report. 
Medical Ethics Committee of the Hospital of Hematology, Chinese Academy of Medical 
Sciences (Institute of Hematology, Chinese Academy of Medical Sciences) 
Contact: Yali Zhang Tel: 022-23909095; E-mail: ec@ihcams.ac.cn 
Address: No. 288 Nanjing Road, Tianjin, China 
 
10 Data Management 
This study used the Case report form (CRF) for data collection. A dedicated person was 
developed to enter the contents of the CRF into the database, and a dedicated person verified 
the consistency of the database with the CRF data. The data manager checked the data in the 
database according to the clinical study protocol, and if there were any questions, a 
questionnaire was filled out and the questions in the questionnaire were answered by the 
investigator. 
 
11 Statistical Methods 
11.1 Sample size 
The primary endpoint of this study was the rate of novel coronavirus infection in healthy 
volunteers. The historical novel coronavirus infection rate was estimated to be about 35%, and 

 
11 
the experimental group was expected to control the infection rate at about 20%, taking a two-
sided α=0.05 with a test efficacy of 90%, and a sample size of 100 cases was calculated using 
a one-sample rate difference z-test continuity correction method, taking into account the 20% 
shedding rate and personnel position stratification factors, and 130 healthy volunteers were 
expected to be enrolled in the final study. The allocation ratios by personnel position were as 
follows: physicians: 20; nurses: 30; medical technicians: 20; managers: 20; researchers: 20; 
and medical students: 20. 
11.2 Statistical analysis set 
Full analysis dataset (FAS): is a dataset determined according to the Intention-To-Treat (ITT) 
principle and includes all enrolled healthy volunteers who have used the study drug at least 
once and have at least one analyzable efficacy indicator, and validity analysis is based on the 
FAS. 
Protocol-compliant dataset (PPS): a subset of the overall FAS, including healthy volunteers 
who did not show significant protocol deviations in the full analysis set, used as an adjunct to 
the validity analysis. 
Safety data set (SS): all healthy volunteers enrolled who received the trial drug and had at least 
one post-dose safety evaluation data, safety analysis was based on SS. 
11.3 Statistical analysis methods 
SAS9.4 software was used for statistical analysis of the data in this study, and all tests 
were performed by two-sided test with test level α=0.05 if not otherwise specified. 
① Basic methods 
The measurement data were described by mean±standard deviation (Mean±SD) or 
median, maximum, minimum, and interquartile range (Median, Max, Min, IQR); the count 
data were counted as number of cases and described by rate or percentage; the comparison of 
measurement data was performed by t test or nonparametric test; the comparison of count data 
was performed by χ2 test or Fisher's exact probability test. 
② Analysis of primary endpoint indicators 
The number and percentage of healthy volunteers with new coronavirus infection were 
calculated, and 95% exact confidence intervals were calculated using the Clopper-pearson 
method. 

 
12 
③ Analysis of secondary endpoint indicators 
The χ2 test or Fisher's exact probability test was used to compare the proportion of neo-
coronavirus-infected patients in different subgroups who turned to severe disease and the 
proportion of neo-coronavirus-infected patients without symptoms, and to calculate the 
confidence interval respectively; the number of days describing the duration of symptoms 
such as fever and sore throat in infected patients and the number of days to turn negative were 
calculated; the antibody status of all healthy volunteers and neo-coronavirus-infected patients 
were described separately. 
④ Analysis of safety indicators 
The occurrence of adverse events AE and serious adverse events SAE were recorded and 
described, and the relevance of the events to the study was determined. 
⑤ Other analyses 
Subgroup and stratified analyses were performed on people of different genders, age 
groups, staff positions, and staff sections to explore the relationship between the levels of 
different factors and the effects of neo-coronavirus infection。 
 
12 Ethics Review 
12.1 Ethical requirements 
This study was conducted in accordance with the current Declaration of Helsinki (2013), 
relevant regulations and ethics committee review. 
Prior to the start of the study, the investigator shall obtain written approval of the study protocol, 
informed consent form, healthy volunteer recruitment procedures, and other written data to be 
provided to healthy volunteers from the relevant regulatory unit in accordance with regulations. 
During the study, if there are any new amendments to the study protocol, informed consent 
form, etc., written approval from the relevant regulatory authorities should be obtained again 
in accordance with the regulations 
12.2 Informed Consent 
The investigator or his/her designated representative will be responsible for explaining to 
each health volunteer or witness the background of the study, the pharmacological 
characteristics of the study medical technique, the study protocol, and the benefits and risks of 

 
13 
participation in the study, and shall obtain written informed consent signed by the health 
volunteer and the study physician prior to the health volunteer's entry into the study (prior to 
the screening examination). 
The final informed consent form should include the following: the purpose of the study, 
the study procedures, the obligations of the health volunteer, the foreseeable benefits and 
foreseeable risks and inconveniences to the health volunteer of participating in the study, the 
availability of treatment for the health volunteer in the event of study-related harm, access to 
study data, and confidentiality of the health volunteer's information. 
The informed consent form should be approved in writing by the relevant regulatory unit 
in accordance with regulations and written in a language that can be read by the healthy 
volunteers. The informed consent form should be signed and dated by the health volunteer, the 
investigator performing the informed consent process, or their representative. One original copy 
of the informed consent form should be retained by the investigator and one by the healthy 
volunteer. If important new data concerning the study are discovered, the informed consent 
form must be revised in writing and sent to the relevant management unit for approval before 
informed consent is obtained again. 
For healthy volunteers who cannot participate in the informed consent process, written 
informed consent must be obtained from the healthy volunteers and all study procedures, etc., 
must be explained to the healthy volunteers. 
12.3 Confidentiality of information 
It is the responsibility of the investigator to maintain the anonymity of the healthy 
volunteers. Only capital letters, numbers, and/or codes may be used to identify healthy 
volunteers on case report forms or other documents, not the names of healthy volunteers. The 
investigator must maintain the healthy volunteer enrollment form that records the healthy 
volunteer's code, name, and home address. The investigator must maintain strict confidentiality 
of documents that reveal the identity of the healthy volunteer. 
 
13Research Management 
13.1 Training 
Before the start of a clinical study, investigators should receive training on the study 
protocol, familiarize themselves with and understand the contents of this clinical study protocol, 
unify the recording methods and judgment criteria, and strictly follow the protocol. 
 

 
14 
13.2 Quality control and assurance 
All observed results and abnormal findings in clinical studies should be carefully verified 
and recorded in a timely manner to ensure the reliability of the data. The investigator enters the 
information required by the protocol into the case report form, and the supervisor verifies that 
it is completed completely and accurately. 
 
14 Confidentiality and publication of research results 
The researcher should keep the information and data related to this study confidential, and 
should not cite or publish the relevant research results or information without the consent of the 
research unit 
 
15 References 
[1] Brevini, T., Maes, M., Webb, G.J. et al. FXR inhibition may protect from SARS-CoV-2 
infection by reducing ACE2. Nature (2022). https://doi.org/10.1038/s41586-022-05594-0 
 
  
 

 
15 
16 Annexes 
Study Informed Consent Form

A single-center, open, single-arm clinical study on the application of ursodeoxycholic acid for the prevention 
of novel coronavirus infections 
Version number：1.0；Version Date：2022-12-15 
Informed Consent Form - Notice Page 
Dear Volunteers. 
We are planning to conduct a single-center, open, single-arm clinical study of the 
application of ursodeoxycholic acid for the prevention of novel coronavirus infection 
at the Hospital of Hematology, Chinese Academy of Medical Sciences (Institute of 
Hematology, Chinese Academy of Medical Sciences) (hereinafter referred to as 
"IHCAMS"), and you will participate in this study as a volunteer. 
Please read the following carefully so that you can make a decision, which is 
entirely your own. 
I. Background and purpose of the study 
In an article titled "FXR inhibition may protect from SARS-CoV-2 infection by 
reducing ACE2" published in Nature on December 5, 2022, ursodeoxycholic acid 
(UDCA) was found to block novel coronavirus entry into human cells by 
downregulating angiotensin-converting enzyme 2 (ACE2) expression through 
inhibition of the transcription factor FXR (from SARS-CoV in 2003 to the SARS-CoV-
2 spectrum in 2019 utilized ACE2 as the primary invasion receptor). Because 
ursodeoxycholic acid acts on the host cell and not on the virus itself, the drug may 
potentially be able to prevent current novel coronavirus and neo-mutant infections, as 
well as infections by other coronaviruses that utilize ACE2. 
This study was conducted because we are currently in a period of high prevalence 
of novel coronavirus infections and there is an urgent need to explore therapeutic 
strategies to prevent neo-coronavirus infections and to prevent further infections in 
medical personnel. 
 
II. Research Procedures 
The study plans to recruit 130 health care workers who are not yet infected with 
novel coronavirus, including doctors, nurses, technicians, administrators and medical 
students, aged 18-65 years old, of any gender, and requiring a negative nucleic acid or 
antigen test for new coronavirus within 48 hours. 
After signing the informed consent form, the researcher will collect basic 
information and perform a routine examination, and collect 8 ml of your blood for 
storage. 
You will then receive ursodeoxycholic acid capsules (250 mg/capsule) for 
infection prevention, 2 capsules per day for 4 weeks; 8 ml of your blood will be 
collected and stored at week 4, week 8 or if infection occurs during the study. 

A single-center, open, single-arm clinical study on the application of ursodeoxycholic acid for the prevention 
of novel coronavirus infections 
Version number：1.0；Version Date：2022-12-15 
You will be followed for 4 weeks after the end of the study, during which time 
you will be continuously monitored for new nucleic acid/antigen status and antibody 
testing.。 
 
III. Possible risks 
All drugs and treatments can cause adverse reactions, and there may be risks 
involved in the research process. Therefore, please be aware of the following details, 
but not everyone who uses the medication will experience these adverse reactions 
below. 
The main effect of ursodeoxycholic acid is to increase the secretion of bile acids 
and to cause changes in the composition of bile, lowering cholesterol and cholesterol 
lipids in bile and facilitating the gradual dissolution of cholesterol in gallstones, which 
is used clinically for gallbladder cholesterol stones and cholestatic liver disease. Its 
adverse reactions are mainly diarrhea, the incidence of which is about 2%; occasionally 
constipation, stomach pain, pancreatitis, headache, dizziness, allergy, bradycardia, etc.; 
skin may be pruritic, hair loss and the occurrence of bronchitis, cough, pharyngitis and 
other adverse reactions of the respiratory system. 
In addition, some if the known risks associated with blood collection are pain, 
burning sensation or bruising at the site of blood collection. If you experience any 
discomfort, or new changes such as infection with a new crown, or any other 
unexpected situation, whether or not related to this study, please contact the investigator 
promptly and they will make a judgment and medical treatment for it.。 
 
IV. Benefits and significance of the study 
Participation in this study may prevent you from contracting a novel coronavirus 
(SARS-CoV-2) or may reduce the symptoms associated with your new coronavirus 
infection, but it may not have any effect. We hope to use this study to explore treatment 
strategies to prevent new coronavirus infection. 
 
V. Confidentiality of personal data 
IHCAMS will strictly protect the privacy and personal information of volunteers 
and will adopt the following protection methods: (1) establish confidentiality measures 
and information security systems, and manage all collected samples anonymously 
(coding management and/or deletion of all identifying information); (2) store samples 
and data securely and set access rights to all samples and/or data; (3) samples and 

A single-center, open, single-arm clinical study on the application of ursodeoxycholic acid for the prevention 
of novel coronavirus infections 
Version number：1.0；Version Date：2022-12-15 
personal information will not be provided to any researchers or institutions outside the 
Blood Institute; (4) data obtained from research may be published or released publicly, 
but your name or personally identifiable information will not be released。 
 
VI. Right to make autonomous decisions 
You have the right to refuse to participate in the Study, and such refusal will not 
affect any other rights or interests you may have or affect your normal work. You may 
withdraw unconditionally at any time 
 
VII. Contact 
If during the study you have any complaints or concerns or question your rights as 
a volunteer, you may contact the following ethics committee staff [Ethics Committee, 
Hospital of Blood Diseases, Chinese Academy of Medical Sciences, Tel: 022-
23909095]. 
 
Commitment of the study physician 
I,as the study physician, confirm that I have clearly explained to the volunteer the 
details of this study, that this study is an over-indication of the drug, including its 
rights as well as possible risks, and have given him/her a signed copy of the 
informed consent form. 
Signature of the study physician: ＿＿＿＿＿  
Date of study: ______________, ______________, ______________ 
Telephone number: ＿＿＿＿＿＿＿＿＿ 
 
Volunteer promise 
I have read and understood the relevant information about this study on the 
informed information page and have had the opportunity to ask questions about 
it. I understand the researcher's explanations. 
I am aware of the risks and benefits that may arise from participating in this 
study. I understand that participation in the study is voluntary and I acknowledge 
that I have had sufficient time to consider and voluntarily participate in this study. 
I have decided to agree to participate in this study. I will receive a signed and 
dated copy of the informed consent form.。 
 
Volunteer's signature: ＿＿＿＿＿＿  

A single-center, open, single-arm clinical study on the application of ursodeoxycholic acid for the prevention 
of novel coronavirus infections 
Version number：1.0；Version Date：2022-12-15 
______ Month ______ Year ______ Day ______ 
Contact number: ＿＿＿＿＿＿＿