Effects of a Marine Protein Hydrolysate in Healthy Adults

Study Overview
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Brief Summary
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This randomized, double blind, controlled trial investigates changes in the cardiovascular index (triacylglycerol/HDL-cholesterol × waist/hip ratio) after 12 weeks of marine protein hydrolysate (MPH) or whey protein powder (placebo) supplementation in adult healthy persons. Additionally, the study investigates potential effects on plasma parameters of metabolic health including lipids, glucose, inflammatory parameters and redox state, as well as associations between dietary MPH and body weight, abdominal obesity, body composition, and gut microbiota composition. Finally, putative end-products of diet-microbial interactions (TMAO and short-chain fatty acids) with CVD risk factors and biomarkers of mitochondrial function are examined.

Detailed Description
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Marine protein sources, including fish and fish protein hydrolysates, may have particular health benefits. Health benefits from fish consumption have been attributed to the n-3 polyunsaturated fatty acids, in particular eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Proteins from marine sources may contain valuable bioactive components, with amino acid composition and protein profiles that differ from terrestrial sources. Generally, the dietary source of protein can affect cellular energy metabolism, and hydrolyzed peptides can have potent and specific bioactive potential. Rest raw materials (RRM) from cod (Fjordlaks AS, Norway) are used for the hydrolysis production (Food Grade). Freshly minced RRM were treated with enzymes optimized to generate bioactive hydrolysates using facilities and techniques approved for human consumption. The investigational products are given in a dose of 18 g protein per day, corresponding to the protein content of a standard meal, and similar to doses recommended in a range of protein supplements. The study enrolls around 70 men and women age of 20-80 years with waist circumference of < 102 cm for men and < 88 cm for women. Prospective study participants were informed of the study and invited to participate through advertising primarily in social media (Facebook advertisement, geographically limited to 12 km surrounding the city centers of Bergen and Ålesund). Participants provided written informed consent, and were screened via self-reporting in an online form in EasyTrial hosted by the Research Unit for Clinical Trials at the University of Bergen. Data collection by the study staff at baseline verifies inclusion and exclusion criteria and participant eligibility prior to randomization. The potential participants are informed about practical details at a digital or physical meeting 1-2 weeks prior to baseline. Groups of participants (40-60% males/females) are block randomized to the two treatments using randomly selected block sizes, and stratified according to sex. The participants are given a container with the powder sufficient for the entire study period, and a spoon to measure the intake at breakfast (6 g), lunch (6 g) and supper (6 g), or morning (9 g) and evening (9 g) according to individual preference. The patients will mix the powder products in water or mineral water. Flavours have been masked by supplementation with beet powder, and mixing 0.5 g fish hydrolysate per serving in the placebo, to minimize differences in taste of the placebo and active product.

Official Title
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Randomized, Double Blind, Placebo Controlled Clinical Study to Investigate the Effect of a Marine Protein Hydrolysate in Healthy Adults

Conditions
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Healthy Diet

Intervention / Treatment
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* Dietary Supplement: Cod protein hydrolysate
* Dietary Supplement: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy female and male subjects at age 20-80 years old understanding Norwegian oral and written information Waist circumference for males < 102 cm and females < 88 cm Exclusion Criteria: Pregnancy or lactation Having used high-dose omega-3 PUFA supplements (>2 g/day) less than 28 days prior to randomization Use of corticosteroids that will influence protein metabolism Antibiotic treatment previous 3 months Alcohol or drug abuse or any conditions associated with poor compliance Medical diagnosis of malabsorption disorders, Crohn's disease, or lactose intolerance Scheduled hospitalization during the course of the study that could compromise the study Diabetes type I or II, serious high blood pressure at screening, serious infections, diseases requiring medication that can influence the study Known or suspected sensitivity or allergic reactions to the IMP or excipients Presence of other major medical or psychiatric illness that would affect the ability to participate in the study or put the subject at increased risk Intake of statins. If needed to obtain the recruitment goals, we will accept people using a low dose of statin

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 80 Years

Sexes Eligible for Study
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All

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: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Randomized, double blind, placebo controlled clinical study
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Placebo (whey protein supplement)<br>Participants will consume 18 g of the placebo whey protein supplement each day for 12 weeks. | Dietary Supplement: Placebo<br>* Whey protein powder, commercially available<br>* Other names: Whey protein powder;|
| Active Comparator: Cod protein hydrolysate supplement<br>Participants will consume 18 g of the cod protein supplement each day for 12 weeks. | Dietary Supplement: Cod protein hydrolysate<br>* Rest raw materials (RRM) from cod (Fjordlaks AS, Norway) are used for the hydrolysis production (Food Grade). Freshly minced RRM is treated wth enzymes optimized to generate bioactive hydrolysates using facilities and techniques approved for human consumption.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in the cardiovascular index (triacylglycerol/high density lipoprotein (HDL)-cholesterol × waist/hip ratio) | Triacylclygerol and HDL-cholesterol concentrations will be measured in serum. Waist and hip circumference will be measured using anthropometric tape over light clothing. For waist circumference, the minimum circumference between the iliac crest and the rib cage will be used. For waist circumference the circumference at the level of the greatest protrusion of the buttocks is used. | Baseline to 12 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in the Quick1 index, a surrogate marker of insulin sensitivity | Measured in serum and calculated as 1 / (log(fasting insulin μU/mL) + log(fasting glucose mg/dL) | Baseline to 12 weeks | 
| Changes in fasting insulin | Measured in serum | Baseline to 12 weeks | 
| Changes in fasting insulin C-peptide | Measured in serum | Baseline to 12 weeks | 
| Changes in fasting glucose | Measured in serum | Baseline to 12 weeks | 
| Changes in total cholesterol | Measured in serum | Baseline to 12 weeks | 
| Changes in non-high density lipoprotein (HDL) cholesterol | Measured in serum | Baseline to 12 weeks | 
| Changes in non-esterified fatty acids (NEFA) | Measured in serum | Baseline to 12 weeks | 
| Changes in triacylglycerol (TAG) | Measured in serum | Baseline to 12 weeks | 
| Changes in gut microbiota composition | Measured by 16S sequencing | Baseline to 12 weeks | 
| Changes in short chained fatty acids (SCFA) | Faecal SCFA concentrations calculated as (mmol/L) × wet faecal weight x faecal moisture content (g/100 g) × 10 | Baseline to 12 weeks | 
| Changes in fat mass/fat free mass ratio | Body composition measured by bioimpedance analysis | Baseline to 12 weeks | 
| Changes in waist-hip ratio | Waist and hip circumference will be measured using anthropometric tape over light clothing. For waist circumference, the minimum circumference between the iliac crest and the rib cage will be used. For waist circumference the circumference at the level of the greatest protrusion of the buttocks is used | Baseline to 12 weeks | 
| Changes in waist-to-height ratio (WHtR) | Waist circumference will be measured using anthropometric tape over light clothing. Hight will be measured using a stadiometer. For waist circumference, the minimum circumference between the iliac crest and the rib cage will be used. | Baseline to 12 weeks | 
| Changes in blood pressure | Blood pressure measurement will be performed manually by a trained nurse using standard equipment | Baseline to 12 weeks | 
| Changes in heart rate | Heart rate will be determined manually by a trained nurse | Baseline to 12 weeks | 
| Changes in glucagon-like peptide 1 (GLP-1, hormone involved in appetite and metabolism regulation) | Measured in plasma by ELISA (enzyme-linked immunosorbent assay) | Baseline to 12 weeks | 
| Changes in gastric inhibitory polypeptide (GIP, hormone involved in metabolism regulation) | Measured in plasma by ELISA (enzyme-linked immunosorbent assay) | Baseline to 12 weeks | 
| Changes in ghrelin (hormone involved in appetite regulation) | Measured in plasma by ELISA (enzyme-linked immunosorbent assay) | Baseline to 12 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Protein supplementation, Cardiovascular disease risk, Metabolic health