Tributyrin Treatment in Mild Alzheimer Disease: Assessment of Butyric Acid Effects Via the Gut-Brain

Phase 3

Not yet recruiting

• Conditions: Alzheimer Disease (AD)

• Registration Number: NCT06797817
• Lead Sponsor: Universidad de Almeria

Brief Summary

The goal of this clinical trial is to learn if tributyrin can help prevent or mitigate cognitive decline in individuals with mild Alzheimer's disease (AD). The trial will also examine the safety and effects of tributyrin on inflammation and gut microbiota. The main questions it aims to answer are:

Does tributyrin improve cognitive function in patients with mild AD? Does tributyrin reduce inflammation and oxidative stress? How does tributyrin affect gut microbiota and intestinal permeability? Researchers will compare tributyrin to a placebo (a look-alike substance that contains no active ingredient) to evaluate its effectiveness.

Participants will:

Take tributyrin or a placebo every day for 12 weeks. Undergo assessments of cognitive function, blood markers (such as BDNF and GFAP), and gut health.

The findings are expected to provide insight into the potential of tributyrin as a preventive intervention for Alzheimer's disease.

Detailed Description

Not available

Study Timeline

• Status Verified: 2025-01
• Study First Submitted: 2025-01-21
• Study First Submitted QC: 2025-01-21
• Study First Posted: 2025-01-28
• Last Update Submitted: 2025-02-06
• Last Update Posted: 2025-02-10
• Study Start: 2025-09-01
• Primary Completion: 2026-09-01
• Study Completion: 2027-09-02

Recruitment & Eligibility

• Status: NOT_YET_RECRUITING
• Sex: All
• Target Recruitment: 156

Inclusion Criteria

• individuals diagnosed with mild AD within the past year (ICD-10: F00.1).
• voluntary consent to participate in the study in accordance with the Declaration of Helsinki.
• not currently enrolled in any other clinical trial that could confound the results.

Exclusion Criteria

• individuals with other potential causes of dementia, such as a history of severe traumatic brain injury, brain tumours, epilepsy, or central nervous system infections.
• individuals involved in an intervention that interferes with the trial (immunosuppressive drugs, steroids, antibiotics, or received chemotherapy in the month prior to the start of the intervention).
• individuals with gastrointestinal disorders.

Study & Design

• Study Type: INTERVENTIONAL
• Study Design: PARALLEL

Primary Outcome Measures

Name	Time	Method
Montreal Cognitive Assessment (MoCA)	Change from Baseline to 12 weeks and 24 weeks	For cognitive evaluation, the Montreal Cognitive Assessment (MoCA) will be utilized, a widely recognized tool developed to identify mild cognitive impairment. In comparison to the Mini-Mental State Examination (MMSE), MoCA exhibits greater sensitivity, particularly in the early stages of AD, allowing for the detection of deficits in areas such as attention, memory, language, abstraction, and executive functions; The MoCA is assessed on a scale ranging from 0 to 30, where higher scores reflect superior cognitive performance. A score of 26 or higher is typically regarded as within the normal range, whereas lower scores indicate different levels of cognitive impairment.

Secondary Outcome Measures

Name	Time	Method
Visual memory by Rey Test	Change from Baseline to 12 weeks and 24 weeks	A comprehensive assessment of neurocognitive abilities will be conducted using neurocomputational tools through the PEBL (Psychology Experiment Building Language) platform. The Rey Test, administered via PEBL, will be used to evaluate both verbal memory and visual memory, providing an integrated measure of episodic memory. Visual Memory: The Rey Test, applied through PEBL, will assess immediate recall (scores from 0 to 15) and delayed recall (scores from 0 to 30), with higher scores reflecting better visual memory performance.
Accuracy in Go/No-Go test (Inhibition)	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. The Go/No-Go test will evaluate inhibition, measuring the ability to control impulsive responses to certain stimuli. Accuracy will be measured by accuracy (the percentage of correct responses), with scores ranging from 0% to 100%. Higher values indicate better control over inhibition.
Reaction Time in N-back Task (Working Memory)	Change from Baseline to 12 weeks and 24 weeks	Participants' reaction time during the N-back task will be measured in milliseconds, using the PEBL platform. Shorter reaction times generally indicate more efficient cognitive processing.
Verbal memory by Rey Test	Change from Baseline to 12 weeks and 24 weeks	A comprehensive assessment of neurocognitive abilities will be conducted using neurocomputational tools through the PEBL (Psychology Experiment Building Language) platform. The Rey Test, administered via PEBL, will be used to evaluate both verbal memory and visual memory, providing an integrated measure of episodic memory. Scores for immediate recall range from 0 to 15, and from 0 to 30 for delayed recall, with higher scores indicating better verbal memory performance.
Accuracy in N-back Task (Working Memory)	Change from Baseline to 12 weeks and 24 weeks	To assess working memory, participants will perform the N-back task (in its 3-4 task versions) using the PEBL (Psychology Experiment Building Language) platform. Performance will be measured by accuracy, calculated as the percentage of correct responses, with higher percentages indicating better working memory performance. Accuracy will be recorded on a scale from 0% to 100%.
Reaction time in Stroop Test (Attention)	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. The Stroop Test will be applied to examine attention, specifically the ability to inhibit automatic responses and manage attention in the presence of conflicting stimuli. Reaction Time will be measured by reaction time (in milliseconds), with shorter times indicating better cognitive control.
Accuracy in Stroop Test (Attention)	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. Accuracy, measured through the Stroop Test will be recorded as the percentage of correct responses, ranging from 0% to 100%, with higher percentages reflecting better performance.
Completion Time in KEFS test (Cognitive flexibility)	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. To assess cognitive flexibility, the D-KEFS will be used, measuring the capacity to switch between different strategies and adapt to new demands. Completion Time administered through PEBL will be measured by completion time (in seconds), with shorter times indicating better cognitive flexibility.
Accuracy in KEFS test (Cognitive flexibility)	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. To assess cognitive flexibility, the D-KEFS will be used, measuring the capacity to switch between different strategies and adapt to new demands. Accuracy measured through the D-KEFS will be recorded, with higher accuracy reflecting better cognitive flexibility.
Reaction time in Go/No-Go test (Inhibition)	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. The Go/No-Go test will evaluate inhibition, measuring the ability to control impulsive responses to certain stimuli. Reaction time will also be assessed in the Go/No-Go test. Faster responses typically indicate more efficient inhibition of impulses. Reaction time may vary depending on the specific version of the task.
Executive functions and impulsivity	Change from Baseline to 12 weeks and 24 weeks	To conduct a comprehensive assessment of neurocognitive abilities, neurocomputational tools will be employed through the PEBL (Psychology Experiment Building Language) platform, which offers a variety of tests designed to measure different aspects of cognition. The Cambridge Gambling Test (CGT) will be utilized to assess decision-making and risk-taking behaviours, providing insights into the participants' executive functions and impulsivity. Performance is generally evaluated by accuracy (the percentage of correct choices) and the inclination to take risks, where greater accuracy indicates stronger decision-making skills. Risk-taking behavior is measured by the amount of money wagered, with larger wagers indicating a higher tendency to take risks.
Neuropsychiatric Inventory Questionnaire (NPI-Q)	Change from Baseline to 12 weeks and 24 weeks	To obtain a comprehensive understanding of the clinical manifestations associated with mild AD, in addition to the neurocognitive tools mentioned, the Neuropsychiatric Inventory Questionnaire (NPI-Q) will be employed. This tool allows for the identification and assessment of the severity of common neuropsychiatric symptoms in this population, such as agitation, depression, anxiety, apathy, and disinhibited behaviors, as well as the level of distress these symptoms cause to caregivers. The severity of each symptom is typically rated on a scale from 0 to 12, with higher scores indicating more severe symptoms. Caregiver distress is measured on a scale from 0 to 5, where higher scores reflect greater distress.
Financial Management in IADL assessment (Independence)	Change from Baseline to 12 weeks and 24 weeks	The Instrumental Activities of Daily Living (IADL) assessment will be included to gain a comprehensive understanding of how mild AD affects daily functioning, as cognitive decline often results in real-life functional impairments. By evaluating abilities such as financial management, meal preparation, medication adherence, and transportation use, this tool provides crucial insights into a patient's level of independence. The financial management component of the IADL assessment evaluates an individual's ability to handle monetary responsibilities independently. A score of 1 indicates that the person can manage finances without assistance, including paying bills on time, budgeting expenses, and keeping track of financial transactions. A score of 0 signifies that the individual is unable to manage finances independently and requires help with banking, bill payments, or financial decision-making. This score contributes to the total IADL score, which ranges from 0 to 8, with higher scores
Meal preparation in IADL assesment (independence)	Change from Baseline to 12 weeks and 24 weeks	The Instrumental Activities of Daily Living (IADL) assessment will be included to gain a comprehensive understanding of how mild AD affects daily functioning, as cognitive decline often results in real-life functional impairments. By evaluating abilities such as financial management, meal preparation, medication adherence, and transportation use, this tool provides crucial insights into a patient's level of independence. The meal preparation component of the IADL assessment evaluates an individual's ability to plan and cook meals independently. A score of 1 indicates that the person can prepare full meals without assistance, including using kitchen appliances safely, cooking properly, and organizing ingredients. A score of 0 signifies that the individual is unable to prepare meals independently and requires assistance for meal planning, cooking, or both. This score contributes to the total IADL score, which ranges from 0 to 8, with higher scores reflecting greater overall independence
Taking medication in IADL assesment (independence)	Change from Baseline to 12 weeks and 24 weeks	The Instrumental Activities of Daily Living (IADL) assessment will be included to gain a comprehensive understanding of how mild AD affects daily functioning, as cognitive decline often results in real-life functional impairments. By evaluating abilities such as financial management, meal preparation, medication adherence, and transportation use, this tool provides crucial insights into a patient's level of independence. The taking medication component of the IADL assessment evaluates an individual's ability to manage their medications independently. A score of 1 indicates that the person can take the correct medications at the right times and dosages without assistance. A score of 0 signifies that the individual requires reminders or full assistance to take medications as prescribed. This score contributes to the total IADL score, which ranges from 0 to 8, with higher scores reflecting greater overall independence in daily activities.
Systemic inflammation	Change from Baseline to 12 weeks and 24 weeks	Due to BA has demonstrated the ability to inhibit pro-inflammatory cytokines while promoting the production of anti-inflammatory cytokines as well as reduce oxidative stress markers. The levels of IL-1β, IL-6, TNF-α, IL-10 cytokines. Blood samples will be collected from participants to assess the mentioned cytokines at baseline, at the end of the 12-week intervention, and three months after its conclusion using ELISA kits following the manufacturer's instructions. Blood samples will be collected at three points. After the sample collection, they will be centrifuged at 2000 x g for 15 minutes, the serum will be obtained, and the blood samples will be stored at -80°C for subsequent analysis. Cytokine levels will be quantified in pg/mL.
Serum MDA levels	Change from Baseline to 12 weeks and 24 weeks	The levels of malondialdehyde (MDA, nmol/mL) will be measured to evaluate oxidative stress. Blood samples will be collected from participants at the same three time points: baseline, the end of the 12-week intervention, and three months after its conclusion. MDA levels will be determined using ELISA kits (Thermo Fisher, California, USA) according to the manufacturer's instructions. Following sample collection, the blood will be centrifuged at 2000 x g for 15 minutes, the serum will be obtained, and the samples will be stored at -80°C until further analysis.
Using transportation in IADL assesment (independence)	Change from Baseline to 12 weeks and 24 weeks	The Instrumental Activities of Daily Living (IADL) assessment will be included to gain a comprehensive understanding of how mild AD affects daily functioning, as cognitive decline often results in real-life functional impairments. By evaluating abilities such as financial management, meal preparation, medication adherence, and transportation use, this tool provides crucial insights into a patient's level of independence. The using transportation component of the IADL assessment evaluates an individual's ability to travel independently. A score of 1 indicates that the person can use public transportation, drive, or arrange their own transportation without assistance. A score of 0 signifies that the individual requires help to travel, whether due to cognitive, physical, or safety concerns. This score contributes to the total IADL score, which ranges from 0 to 8, with higher scores reflecting greater overall independence in daily activities.
Serum Levels of BDNF	Change from Baseline to 12 weeks and 24 weeks	Elevated levels of BDNF are associated with improved cognitive function. In this sense and given the anti-inflammatory properties of BA, this study aims to investigate whether tributyrin treatment can modulate the BDNF levels. To perform this, serum BDNF levels (pg/mL) will be measured using appropriate ELISA kits (Thermo Fisher, California, USA) following the manufacturer's instructions. After the sample collection, they will be centrifuged at 2000 g for 15 minutes, the serum will be obtained, and it will be stored at -80 °C until further analysis.
Serum SOD levels	Change from Baseline to 12 weeks and 24 weeks	Due to BA having demonstrated the ability to reduce oxidative stress markers, the activity of superoxide dismutase (SOD, U/mL) will be analyzed. Blood samples will be collected from participants to assess SOD activity at baseline, at the end of the 12-week intervention, and three months after its conclusion using ELISA kits (Thermo Fisher, California, USA) following the manufacturer's instructions. After the sample collection, they will be centrifuged at 2000 x g for 15 minutes, the serum will be obtained, and the blood samples will be stored at -80°C for subsequent analysis.
Concentration of Tight Junction Proteins	Change from Baseline to 12 weeks and 24 weeks	To assess whether tributyrin treatment enhances tight junction protein expression, reduces inflammation, and partially restores barrier function, faecal levels of tight junction proteins. Occludin and ZO-1 proteins will be measured in the participants' faces at the three time points of the study. Protein levels will be quantified by ELISA using mouse monoclonal antibodies specific for occludin (1:50, ref: OC-3F10) and ZO-1 (1:100, ref: ZO1-1A12), both purchased from Thermo Fisher, California, USA. A secondary antibody, goat anti-mouse IgG, HRP-conjugated (ref: 31430), will be used for detection available from Thermo Fisher. The samples will be stored at -80°C until analysis. Protein levels will be measured in pg/mL.
Serum Levels of GFAP	Change from Baseline to 12 weeks and 24 weeks	GFAP serves as a marker of astrocyte activation and neuroinflammation. In this sense and given the anti-inflammatory properties of BA, this study aims to investigate whether tributyrin treatment can modulate GFAP levels. To perform this, serum GFAP levels (ng/mL) will be measured using appropriate ELISA kits following the manufacturer's instructions. After the sample collection, they will be centrifuged at 2000 g for 15 minutes, the serum will be obtained, and it will be stored at -80 °C until further analysis.
Composition of Intestinal Microbiota (16S rRNA Gene Sequencing)	Change from Baseline to 12 weeks and 24 weeks	To explore the influence of tributyrin supplementation on the composition of the intestinal microbiota, 16S rRNA gene sequencing will be performed on fecal samples from participants before, at the end of the supplementation, and three months after its conclusion. Total bacterial DNA from fresh stool samples will be extracted using the Fast DNA Stool Kit (Thermo Fisher Scientific, California, USA). The 16S rRNA gene will be sequenced using universal primers (341F/806R) to target the V3-V4 region, following methods outlined in previous studies. The raw data will be processed with QIIME2 software, and further bioinformatics analysis will be performed using the MicrobiomeAnalyst online platform (https://www.microbiomeanalyst.ca). This will include assessments of alpha and beta diversity, core microbiome analysis, and random forest modeling.
Levels of SCFAs (Acetic Acid, Propionic Acid, and Butyric Acid) in faeces	Change from Baseline to 12 weeks and 24 weeks	To evaluate whether BA supplementation promotes increased levels of SCFAs, levels of acetic acid (AA), propionic acid (PA), and BA will be measured. Faecal samples will be collected before, after 12 months, and three months post-intervention. Faecal samples will be collected in a home setting using tubes containing a stabilizing medium. The tubes will be hermetically sealed to minimize the risk of SCFAs volatilization and ensure sample integrity. SCFA levels will be quantified using gas chromatography-mass spectrometry (GC-MS) (Agilent Technologies). The units of measure for SCFAs will be expressed in µmol/g of faeces.

Trial Locations

Locations (1)

Universidad de Almería; 🇪🇸 Almeria, Spain