1. Research purpose:
Main Objective: To study the efficacy and safety of allogeneic adipocyclical active
protein in the treatment of severe idiopathic pulmonary fibrosis, and the main
observation and evaluation indicators include: clinical efficacy, clinical adverse
events and laboratory abnormalities.
The basis of the question:
Animal experiments have found that Cell Free Fat Extract (CEFFE) has a certain
improvement effect in bleomycin-induced mouse pulmonary fibrosis model. At present,
domestic clinical studies have found that Cell Free Fat Extract (CEFFE) is safe for the
treatment of human diseases without obvious side effects. Therefore, in this study, the
Cell Free Fat Extract (CEFFE) was inhaled by nebulized inhalation for the treatment of
severe idiopathic pulmonary fibrosis, and its efficacy and safety were observed.
2. Research Design:
Overall design: single-center, self-controlled trial design was adopted, no control
group was established, and blinding was not used.
Number of planned cases: 7.
Indications: severe idiopathic pulmonary fibrosis.
3. Clinical trails process:
3.1 Sign the informed consent form and collect relevant clinical data; For patients with
a definitive diagnosis of severe idiopathic pulmonary fibrosis. 3.2 Allogeneic Cell Free Fat Extract (CEFFE) treatment process:
3.2.1 Preparation of CEFFE of allogeneic origin: Healthy donors undergoing liposuction
at the Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of
Medicine, sign a written informed consent. By a healthy donor under local anesthesia,
inject swelling fluid into the abdomen, operate with a 20mL syringe or mechanical
negative pressure machine (auxiliary liposuction methods such as laser, ultrasound and
microwave cannot be added), determine the group according to the order of enrollment,
and extract fat according to the required dose, and the single liposuction amount should
be 150ml -200ml. The collected adipose tissue was rinsed with normal saline, allowed to
stand, and centrifuged in a centrifuge at 1200g for 3min. After centrifugation, excess
water at the bottom is removed, collected in a sterile 50mL syringe, stored, and
transferred. Preparation of adipose tissue active factors is performed in the laboratory
and cryopreservation. Two 10mL syringes connected by tee tubes, mechanically emulsified
60 times, collected into 50mL centrifuge tubes, centrifuged at 2000g for 5min. After
centrifugation, the bottom transparent liquid layer was collected and filtered through a
0.22um filter to obtain fat decellularization active protein, which was frozen in a -20
°C freezer.
3.2.2 Health Donor Selection Criteria:
3.2.2.1 Age 18-45, male or female. 3.2.2.2 Liposuction at the Ninth People's Hospital Affiliated to Shanghai Jiao Tong
University School of Medicine. 3.2.2.3 Sign the informed consent form for biological sample donation. 3.2.3 Exclusion Criteria:
3.2.3.1 Patients with hepatitis B, hepatitis C, HIV, syphilis, cytomegalovirus,
herpesvirus and other infections. 3.2.3.2 Patients with severe hypertension, diabetes, heart disease and other chronic
diseases. 3.2.3.3 Patients with tumors or previous history of tumors. 3.3 After negative percutaneous test, allogeneic CEFFE is inhaled by nebulization; 0.2ml
CEFFE was injected intradermally 1 cm above the palmar stria on the palmar side of the
right forearm, and 0.2ml of normal saline was injected intradermally 1 cm above the
palmar stria on the palmar side of the left forearm as a control. Interpretation of skin
test results: local pichu bulge, and red halo hard lump, diameter > 1 cm is positive.
Give a single dose of 2ml allogeneic CEFFE stock solution (protein concentration of
3mg/ml), administered by nebulization inhalation, 8 minutes between each atomization
time, 3 days apart; A total of 7 doses were given.
3.4 Observe the clinical symptoms, chest CT, lung function and related adverse reactions
of patients at 1, 3, 6 and December after the end of allogeneic fat decellularization
active protein treatment.
4. Evaluation Criteria:
4.1 Efficacy criteria:
4.1.1 Obvious effect means that the main symptoms and signs are significantly relieved,
and the objective indicators are significantly improved.
4.1.2 Effective means that the main symptoms and signs have improved, and the objective
indicators have improved.
4.1.3 Ineffective means that the main symptoms and signs have not changed, and the
objective indicators have not changed significantly or worsened.
4.2 Safety evaluation:
Clinical adverse events and laboratory tests are definitely related to allogeneic CEFFE
treatment, likely related to allogeneic CEFFE treatment, may be related to allogeneic
CEFFE treatment, may not be related to allogeneic CEFFE treatment, and may not be
related to allogeneic CEFFE treatment.
4.3 Evaluation endpoints: Clinical efficacy, comprehensive efficacy and safety
evaluation at 1, 3, 6 and 12 months after the end of treatment were the main evaluation
endpoints.
5. Quality assurance of clinical trial data. The quality control of this clinical trial is controlled and controlled by the Ninth
People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine, and
the research work is carried out in accordance with GCP, SOP and quality control
requirements. Develop a clinical research plan. GCP training for relevant researchers
participating in the trial.
5.1 Before the start of the trial, the plan should be submitted to the Ethics Committee
of the Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of
Medicine for approval;
5.2 in accordance with GCP guidelines, necessary steps should be taken during the design
and implementation phases of the study to ensure that the data collected is accurate,
consistent, complete and credible;
5.3 The participating researchers will strictly follow China's GCP standards to conduct
the trial, collect and record the contents of the case report form truthfully, carefully
and in detail in accordance with the clinical trial standard operating procedures (SOP),
and verify them to ensure the reliability of the data;
5.4 The investigator fills in the information required by the protocol into the Case
Report Form (CRF), and the supervisor verifies that it is complete and accurate, and
instructs the staff of the research center to make necessary corrections and
supplements;
5.5 All kinds of instruments, equipment, reagents, standards, etc. used in various
inspection items in clinical trials should have strict quality standards and ensure that
they work under normal conditions;
5.6 Send statistical issues in the test plan to statistical experts for review and
check;
5.7 During the trial, the investigator monitors the research process, informed consent,
and the correctness and completeness of the data in the case report form (CRF).
6. Recording and preservation of research materials. In accordance with the GCP principle, the investigator should keep all the detailed
original files of the subjects, and record the relevant test process, medication,
laboratory test data, safety data and efficacy evaluation in the case report form, and
the recorded data should be complete, timely and clear. Case report forms, original
documents, medical records, etc. should be clear, detailed and easily identifiable by
personnel participating in this clinical trial.
At a minimum, the principal investigator must sign the inclusion confirmation page and
completion page of the case report form to confirm the accuracy and completeness of all
data.
The case report form and the original file can only be modified by the investigator. Any
changes to the case report form and the original file must not erase the original data.
The correct modification method is to draw a single line on the original data, and then
write the modified data next to the original data, and sign the date and the initials of
the person who modified it.
Test data should be retained for 10 years after the end of the test. However, if
required by current regulations or agreements with sponsors, these data should be kept
for a longer period of time.
7. statistical treatment scheme. 7.1 filling in the case report form (CRF) The investigator should complete the filling
of case report form in time, modify it according to the correct modification method, and
the completed case report form shall be reviewed and signed by the main researchers. 7.2 statistical analysis method and statistical description. 7.2.1 Treatment of missing value of main efficacy index data: when some subjects lack a
major efficacy data, the method of filling deficiency is determined from the statistical
and professional perspective. If the case is missing, the data of the previous
measurement shall be transferred.
7.2.2 Case analysis of incomplete test: the reasons for exfoliation should be analyzed
one by one.
7.2.3 Descriptive statistics: mean, standard deviation, maximum value, minimum value,
median, confidence interval, frequency (composition ratio), etc.
7.3 Statistical expression. 7.3.1 The report mainly uses tables to show that the tables are self-evident, that is,
they have table questions, table notes and examples.
7.3.2 The results of repeated measurement data are expressed in tables and statistical
charts are attached to increase readability.
7.4 Statistical software SAS 9.1.3 and Das for clinical trial 3.0 software were used to
analyze the software simultaneously, and the results were confirmed.
8. Modification and interim analysis in the course of the test. Once the research plan has been discussed by the research group and reviewed by the ethics
committee, it is not allowed to change at will; before the modification is implemented, the
researcher must report the content and reason of the modification to the ethics committee of
clinical trial for approval, and then carry out the research according to the modified
scheme.
