The Role of Beta-1,3/1,6-D-Glucan From Mycelium Extract of Indonesian Ganoderma Lucidum on Ulcerative Colitis: A Double-Blind Randomized Controlled Trial
Phase: PHASE2
Conditions: Ulcerative Colitis
Sponsor: Fakultas Kedokteran Universitas Indonesia

  
 
  
STUDY PROTOCOL 
(June 30th 2019) 
 
The Role of Beta-1,3/1,6-D-Glucan from Mycelium Extract of 
Indonesian Ganoderma Lucidum on Ulcerative Colitis: A Double-Blind 
Randomized Controlled Trial 
 
 
 
  
 
PRINCIPAL INVESTIGATOR : Prof. dr. Marcellus Simadibrata, PhD., SpPD-KGEH  
CO-INVESTIGATOR 
 
   : Prof. Dr. dr. Murdani Abdullah, SpPD-KGEH 
 
 
 
 
     dr. Rabbinu Rangga Pribadi, SpPD   
Faculty of Medicine 
Universitas Indonesia 
(FMUI) 
   
 
 
 
 
2019  
 
 

 
 
Title 
The Role of Beta-1,3/1,6-D-Glucan from Mycelium Extract of Indonesian Ganoderma Lucidum on 
Ulcerative Colitis: A Double-Blind Randomized Controlled Trial 
Background 
Inflammatory bowel disease (IBD) is an autoimmune disorder there is marked by chronic 
inflammation in gastrointestinal system.1 IBD is classified to two types, ulcerative colitis (UC) and 
Crohn’s disease (CD). These are based on difference pathology and clinical manifestation. The 
common symptom of IBD is chronic diarrhoea with or without blood, abdominal cramp and weight 
loss.2,3 IBD has high incidence and prevalence in Europe and America. Incidence of UC and CD in 
Europe are 24.3 and 29.3 by 100,000 respectively.4 On the last few decades, there was an increasing 
incidence and prevalence IBD on Asia and other developing countries.5 In Indonesia, incidence of 
UC and CD are 0.55 and 0.33 by 100.0000 respectively.6 Trends in epidemiology of IBD was related 
to westernized lifestyle, urbanization, and hygiene as a part of socioeconomic development in 
developing countries.7      
Nowadays, IBD treatment focus to prevent complication, maintain remission phase and improve 
quality of life. Drugs that are widely used for improving symptoms of IBD are anti-inflammatory 
drugs (5-aminosalicylic acid), immunomodulator (azathioprine, mercaptopurine, methotrexate), and 
biologic agent. These drugs work by increasing Th2-mediated response that cause a decrease in 
inflammation mediated by Th1.8 Long term use of these drugs will cause different side effects. Side 
effects from immunomodulator are myelosuppresion, hepatitis and increase risk of malignancy. Use 
of anti TNF-α will cause lupus like syndrome and malignancy.9 Therefore, we need alternative 
treatment of IBD that effective but minimal side effects. 
Ganoderma lucidum is a species from Ganodermataceae family that has Beta-1,3/1,6-D-Glucan as 
one of active compound from isolated mycelium extract Ganoderma lucidum. This compound has 
biologic effect as immunomodulator and has been used to prevent and treat diseases such as gastric 
ulcer, arthritis, atherosclerosis, asthma, diabetes, cancer and other diseases related to immune 
disorders.10,11 A study on patient with CD showed that Ganoderma lucidum decrease TNF-α, IFN-
γ, IL-17A from inflammation of intestinal mucosa.9 On the other hand, there is no study about the 
role of Beta-1,3/1,6-D-Glucan Ganoderma lucidum in UC.  

Objective 
General objective 
To know the role of Beta-1,3/1,6-D-Glucan from mycelium extract of Ganoderma lucidum 
administration in ulcerative colitis 
Specific objectives 
1. To know the quality of life from patient with ulcerative colitis after Beta-1,3/1,6-D-Glucan 
mycelium extract of Ganoderma lucidum administration 
2. To know the level of inflammatory marker from patient with ulcerative colitis after Beta-
1,3/1,6-D-Glucan mycelium extract of Ganoderma lucidum administration 
3. To know the Mayo score from patient with ulcerative colitis after Beta-1,3/1,6-D-Glucan 
mycelium extract of Ganoderma lucidum administration 
 
Methods 
Study Design 
This research is a double blind, randomized clinical trial method. Analysis will be conducted pre 
and post administration: control (placebo) and intervention (polysaccharide peptide Ganoderma 
lucidum) 3 times daily for 90 days. 
Location and Time 
This research will be conducted on Indonesia from August 2019 to December 2020 
Population and Subject 
Population 
The population of this research is patients diagnosed with ulcerative colitis in Indonesia 
 
Subjects Criteria 
- Inclusion criteria 
1. Subjects aged ≥ 18 years old 
2. Ulcerative colitis patients who are treated by 5-ASA 3x500 mg 
3. Agreed to participate 
 
- Exclusion criteria 
1. Allergic with Ganoderma lucidum 

2. Ulcerative colitis patients who are treated by corticosteroid, immunosuppressive agent, 
biologic agent 
3. Could not be randomised and participate in this study by clinical judgement 
Drop Out Criteria 
Drop out subjects are subjects who have signed the informed consent but decide to withdraw before 
research is completed  
 
Estimated Sample Size 
Minimum sample size is determined using the following equation for proportion equation. 
N1 = N2 =
(𝑍𝛼√2PQ + 𝑍𝛽√P1Q1 + P2Q2)2
(P1 −P2)2
 
Description :  
Zα = 1.96  
Zβ = 0.84  
P1-P2 = 0.2 
P1 = 0.55412 
N1 = N2 = 92.25, thus with predicted drop out 10% the minimal sample for each group is 102 
subjects 
 
Randomization  
Randomization is generated by computer and subjects will be given code and divided into two 
groups (control and intervention). Researcher and subjects are blinded to treatment. 
Variables 
Independent Variables 
1. Intervention: administration of capsule contain Beta-1,3/1,6-D-Glucan as active form of 
polysaccharide peptide from mycelium extract Ganoderma lucidum 3 times daily 
2. Control (placebo): administration of empty capsule 3 times daily 
Dependent Variables 
1. Quality of life; assessed by Indonesian validated SF-36 questionnaire 
2. Erythrocyte Sedimentation Rate (ESR), C-Reactive Protein (CRP), Tumor Necrosis Factor 
Alpha (TNF-α) and Interleukin 6 (IL-6) from blood samples 

3. Fecal calprotectin from stool examination 
4. Mayo Score from colonoscopy 
5. Biopsy  
 
Procedure 
1. Eligibility assessment 
The subjects will be assessed for eligibility using inclusion and exclusion criteria. 
2. Subjects sign the informed consent form 
3. Randomization 
4. Pre-administration 
Before administration of polysaccharide peptide or placebo based on randomization, subjects 
will be analysed for his/her quality of life by SF 36 questionnaire, ESR, CRP, TNF-α, IL-6, 
fecal calprotectin, colonoscopy (Mayo Score) and biopsy. 
5. Administration (90 days) 
Subjects will be given Ganoderma lucidum and placebo that will be taken three times daily 
for 90 days. The drugs are given monthly to the subjects.  
6. Compliance monitoring 
Compliance monitoring of administration is conducted by the investigators coordinating with 
his/her family and checked monthly when the patients came to take the capsule for the next 
month 
7. Post administration  
After 90 days the subjects will be analysed for his/her quality of life by SF 36 questionnaire, 
ESR, CRP, TNF-α, IL-6, fecal calprotectin, colonoscopy (Mayo Score) and biopsy. 
8. Data analysis 
a. Normality analysis using Kolmogorov-Smirnov (Normally distributed if p>0.05) 
b. Bivariate analysis by comparing numeric data between two groups: if data is 
normality distributed using T-test, if not using Mann-Whitney test (significant if 
p<0.05) 
c. Bivariate analysis by comparing categorical data between two groups: if data is 
normality distributed using T-test, if not using Mann-Whitney test (significant if 
p<0.05) 

d. Bivariate analysis by comparing numeric data between preadministration and 
postadministration: if data is normality distributed using paired T-test, if not using 
Wilcoxon test (significant if p<0.05) 
e. Bivariate analysis by comparing categorical data between preadministration and 
postadministration: if data is normality distributed using Mc Nemar test, if not using 
Wilcoxon test (significant if p<0.05) 
 
Study flow chart 
Eligibility 
assessment 
Ulcerative colitis patient 
Informed consent 
Randomization 
Intervention group: 
Ganoderma lucidum 
Control group: 
Placebo 
Administration 3 times daily for 90 days 
Day 90   quality of life, CRP, LED, TNF-α, IL-6, fecal calprotectin, Mayo score 
(colonoscopy), biopsy 
Day 0  quality of life, CRP, LED, TNF-α, IL-6, fecal calprotectin, Mayo score 
(colonoscopy), biopsy 

Ethical committee approval  
Approval from the Ethical Committee at the Faculty of Medicine Universitas Indonesia must be 
obtained before starting the trial. 
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