Determination of Biological Activity of Enriched Serum Following TOTUM-448 Consumption

Study Overview
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Brief Summary
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Dietary intakes of saturated fatty acids remain well above nutritional recommendations for most European countries. This may progressively lead to a pro-inflammatory context and the alteration of lipid metabolism in the liver. The hypothesis of this study is that TOTUM-448 promotes the normal function of hepatocytes in a mild inflammatory context. To determine the cellular and molecular effects of TOTUM-448, circulating bioactive molecules after TOTUM-448 intake will be collected and then studied in human hepatocytes using an innovative ex vivo clinical approach developed by Clinic'n'Cell.

Official Title
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Determination of Biological Activity of Enriched Serum Among Healthy Participants After Consumption of the Food Supplement TOTUM-448

Conditions
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MASLD, Mild Inflammatory Context, NAFLD

Intervention / Treatment
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* Dietary Supplement: TOTUM-448


Participation Criteria
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Eligibility Criteria
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Main Inclusion Criteria: Lipid profile within reference values Glycemia < 100mg/dL Normal hepatic function Normal blood formulation Normal renal function BMI between 20 and 28kg/m2 Non smoker or occasional smoker Main Exclusion Criteria: All types of vaccination within one month Alcohol intakes superior to World Health Organization recommendations Ongoing pharmacological, dietary supplement and/or probiotic treatment Consumption of enriched functional foods or dietary supplements within two weeks of inclusion Any known disease Allergy to ingredients of the study product With dietary habits incompatible with the study conduct

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 35 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Intervention Model: Single Group Assignment
Interventional Model Description: An ex vivo clinical approach.
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: TOTUM-448<br>10 healthy men will consume an acute intake of 4.284g of TOTUM-448 on three separate occasions. Phase 1: once per galenic form (capsule and powder). Phase 2: once (capsule only). | Dietary Supplement: TOTUM-448<br>* The study is divided in two phases. The first clinical phase of the project is aimed at determining TOTUM-448's metabolites absorption peak (two galenic forms will be tested: capsules and powder). Then, human circulating metabolites from polyphenols will be quantified and characterized by ultra-high performance liquid chromatography hyphenated with tandem mass spectrometry (UPLC-MS/MS). Once the absorption profile is characterized, volunteers will visit once again the clinical center for the collection of serum fractions either naïve (before ingestion) or enriched (containing circulating metabolites following TOTUM-448 ingestion). The enriched fraction (circulating bioactives collection) will be collected at t-max as characterized in the first phase (absorption profil's peak). Finally, biological activity of TOTUM-448 on human hepatocytes will be studied by comparing naïve serum and serum containing circulating metabolites according to an in vitro protocol.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Evaluate the influence of circulating bioactives' metabolites in blood stream resulting from the intake of TOTUM-448 on viability of human hepatocytes in a hyperlipidic pro-inflammatory context. | Naïve will be compared to enriched serum from the 10 healthy participants at the previously determined t-max. | At phenolic compounds t-max obtained during kinetic phase. Note: According to previous studies, t-max is expected around 60 minutes. | 
| Evaluate the influence of circulating bioactives' metabolites in blood stream resulting from the intake of TOTUM-448 on cholesterol storage of human hepatocytes in a hyperlipidic pro-inflammatory context. | Naïve will be compared to enriched serum from the 10 healthy participants at the previously determined t-max. | At phenolic compounds t-max obtained during kinetic phase. Note: According to previous studies, t-max is expected around 60 minutes. | 
| Evaluate the influence of circulating bioactives' metabolites in blood stream resulting from the intake of TOTUM-448 on triglyceride storage of human hepatocytes in a hyperlipidic pro-inflammatory context. | Naïve will be compared to enriched serum from the 10 healthy participants at the previously determined t-max. | At phenolic compounds t-max obtained during kinetic phase. Note: According to previous studies, t-max is expected around 60 minutes. | 
| Evaluate the influence of circulating bioactives' metabolites in blood stream resulting from the intake of TOTUM-448 on oxidative stress status of human hepatocytes in a hyperlipidic pro-inflammatory context. | Naïve will be compared to enriched serum from the 10 healthy participants at the previously determined t-max. | At phenolic compounds t-max obtained during kinetic phase. Note: According to previous studies, t-max is expected around 60 minutes. | 
| Evaluate the influence of circulating bioactives' metabolites in blood stream resulting from the intake of TOTUM-448 on pro-inflammatory cytokine expression of human hepatocytes in a hyperlipidic pro-inflammatory context. | Naïve will be compared to enriched serum from the 10 healthy participants at the previously determined t-max. | At phenolic compounds t-max obtained during kinetic phase. Note: According to previous studies, t-max is expected around 60 minutes. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Time to reach the maximum concentration in serum of detectable polyphenol metabolite concentrations by UPLC-MS/MS. | After ingestion of TOTUM-448 the concentration in serum of detectable polyphenol metabolites will be assessed every 20 minutes during 240 minutes. | From timepoint 0 minute to timepoint 240 minutes. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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ex vivo, lipid metabolism, inflammation, hepatocytes, oxidative stress, plant extract, polyphenols, dietary/food supplement