Effect of a Multi-ingredient on Visceral Adiposity and Non-alcoholic Fatty Liver Disease in Individuals With Abdominal Obesity

Study Overview
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Brief Summary
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The aim of this study is to validate the efficacy of specific combination of different natural histidine-related amino acids in the reduction of visceral fat and liver steatosis, as well their associated comorbidities, in individuals with abdominal obesity.

Detailed Description
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World Health Organization (WHO) defines obesity as an excess of fat accumulation in adipose tissues and in other metabolic organs, leading to serious health implications. Obesity is a rising issue whose prevalence has defined as a global pandemic or globesity (>30% world population) being responsible of millions of deaths annually. Also, obesity is related with several comorbidities such as non-alcoholic fatty liver disease (NAFLD), with a global prevalence of >25%. Thus, strategies to ameliorate obesity and NAFLD are critical to improve life expectancy and quality of life and to reduce the economic burden of both diseases. Current therapies against obesity are mainly focused on weight loss, including lifestyle intervention, to modify eating behaviours and to promote physical activity. However, the compliance of patients with these therapies is small. In addition, there are some drugs to fight against these diseases. In obesity, these medical therapies are focused to decrease fat gastrointestinal absorption using lipase inhibitors with several side effects: faecal incontinence, abdominal cramping and raise in blood pressure. In NAFLD, medical therapies are designed to reduce insulin resistance, using pioglitazone and metformin. However, the European Medicines Agency (EMA) and the Food and Drug Administration (FDA) recommend avoiding pioglitazone use by its relationship with heart failure and cancer, and metformin provides just a modest improvement in NAFLD. Thus, the finding of new and efficient therapeutic agents is highly desirable to combat these global diseases. The main objective of this study is to evaluate the effect of daily intake of a specific combination of L-histidine, L-serine, L-carnosine and N-Acetylcysteine, in combination with dietary recommendations, on the amount of visceral fat in individuals with abdominal obesity. The secondary objectives of this study are to evaluate the effect of daily intake of the multi-ingredient aforementioned in liver fat content and obesity related comorbidities. Participants who fulfilled the inclusion and exclusion criteria will be randomly assigned to the intervention and control group. During the study there will be 4 visits: a preselection visit (V0; day -7) and 3 study visits during the consumption of the treatments, which will take place on the first day of the study (V1; day 1 +/- 3 days; week 1), at 6 weeks of treatment (V2; day 44 +/- 3 day; week 6) and at 12 weeks of treatment (V3; day 90 +/- 3 days; week 12).

Official Title
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Effect of a Multi-ingredient of L-Histidine, L-Serine, L-Carnosine and N-Acetylcysteine on Visceral Adiposity and Non-alcoholic Fatty Liver Disease in Individuals With Abdominal Obesity. Randomized, Parallel, Placebo Controlled, Triple Blind Study.

Conditions
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Visceral Obesity, Non-Alcoholic Fatty Liver Disease

Intervention / Treatment
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* Dietary Supplement: Multi-ingredient of L-histidine, L-serine, L-carnosine and N-Acetylcysteine
* Dietary Supplement: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Men older than 40 years. BMI ≥30.0 kg/m^2 and ≤35.0 kg/m^2 Waist circumference ≥102 cm. Read, write and speak Catalan or Spanish. Sign the informed consent. Exclusion Criteria: Present values of body mass index > 35 kg/m^2 Present values of waist circumference > 150 cm. Present diabetes. Present dyslipidemia (LDL cholesterol ≥ 189 mg/dL and/or triglycerides ≥ 350 mg/dL). Present anemia. Taking supplements, multivitamin supplements or phytotherapeutic products that interfere with the treatment under study. Consume 4 or more Standard Beverage Units (SBU) daily or 28 SBU weekly. Be a smoker. Present any diagnosed liver disease other than NAFLD. Have lost more than 3 kg of weight in the last 3 months. Present food intolerances and/or allergies related to the study products, such as hypersensitivity to maltodextrin or N-Acetylcysteine. Presenting any chronic or autoimmune disease in clinical manifestation such as hepatitis, hyper or hypothyroidism or metabolic diseases. Follow a pharmacological treatment with immunosuppressants, cytotoxic agents, corticosteroids or other drugs that could cause hepatic steatosis or alter the measurements in the liver. Being participating or having participated in a clinical trial or nutritional intervention study in the last 30 days before inclusion in the study. Follow a hypocaloric diet and/or pharmacological treatment for weight loss. Suffering from eating behavior disorders or psychiatric disorders. Being unable to follow study guidelines.

Ages Eligible for Study
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Minimum Age: 40 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: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Multi-ingredient of L-histidine, L-serine, L-carnosine and N-Acetylcysteine<br>Participants will daily consume the multi-ingredient (L-Histidine, L-Serine, L-Carnosine and N-Acetylcysteine) for 12 weeks. | Dietary Supplement: Multi-ingredient of L-histidine, L-serine, L-carnosine and N-Acetylcysteine<br>* The product will be presented in powder format in a single container and with a measuring spoon of the daily dose.<br>|
| Placebo Comparator: Placebo<br>Participants will daily consume the placebo (maltodextrin) for 12 weeks. | Dietary Supplement: Placebo<br>* The product will be presented in powder format in a single container and with a measuring spoon of the daily dose.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in Visceral Adiposity | Visceral fat content measured using a dual energy x-ray absorptiometry (DXA) scanner | Change from Baseline Visceral Adiposity at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in Hepatic Steatosis | Liver steatosis will be qualitative determined by ultrasound | Change from Baseline Hepatic Steatosis at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Height (cm) | Height measured with standardized method | At Baseline | 
| Change in Weight (kg) | Weight measured with standardized method | Change from Baseline Weight at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Body Mass Index (BMI) (Kg/m^2) | Weight and height will be combined to report BMI in kg/m^2 | Change from Baseline Body Mass Index at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Neck circumference (cm) | Neck circumference using a measuring tape | Change from Baseline Neck circumference at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Arm circumference (cm) | Arm circumference using a measuring tape | Change from Baseline Arm circumference at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Waist circumference (cm) | Waist circumference using a measuring tape | Change from Baseline Waist circumference at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Conicity Index | Weight, height and waist circumference will be combined to report Conicity index. | Change from Baseline Conicity Index at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Systolic Blood Pressure (mm Hg) | Systolic blood pressure will be measured using an automatic sphygmomanometer | Change from Baseline Systolic Blood Pressure at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Diastolic Blood Pressure (mm Hg) | Diastolic blood pressure will be measured using an automatic sphygmomanometer | Change from Baseline Systolic Blood Pressure at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in intestinal microbiota composition | Metagenomic analysis in fecal samples. The bacteria DNA will be extracted and massive sequenced by the Ion Torrent platform. | Change from Baseline intestinal microbiota composition at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change of biomarkers of oxidative stress (8-OHdG, F2-isoprostanes) | Biomarkers of oxidative stress will be evaluated in urine by standardized chemiluminescence methods | Change from Baseline biomarkers of oxidative stress at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum glucose levels (mg/dL) | Serum glucose levels will be determined by standardized spectrophotometry methods | Change from Baseline serum glucose levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum total cholesterol (mg/dL) | Total cholesterol will be determined by standardized spectrophotometry methods | Change from Baseline serum total cholesterol at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum high-density lipoprotein cholesterol (HDL-C,mg/dL) | High-density lipoprotein cholesterol will be determined by standardized spectrophotometry methods | Change from Baseline serum high-density lipoprotein cholesterol at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum low-density lipoprotein cholesterol (LDL-C, mg/dL) | Low-density lipoprotein cholesterol will be calculated using the Friedewald formula | Change from Baseline serum low-density lipoprotein cholesterol at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum triglycerides (TG, mg/dL) | Triglycerides will be determined by standardized spectrophotometry methods | Change from Baseline serum triglycerides at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum alanine aminotransferase (ALT, U/L) | Alanine aminotransferase will be determined by standardized spectrophotometry methods | Change from Baseline serum alanine aminotransferase at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum aspartate aminotransferase (AST, U/L) | Aspartate aminotransferase will be determined by standardized spectrophotometry methods | Change from Baseline serum aspartate aminotransferase at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum gamma glutamyl transferase (GGT, U/L) | Gamma glutamyl transferase will be determined by standardized spectrophotometry methods | Change from Baseline serum gamma glutamyl transferase at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum insulin levels (mU/L) | Insulin levels will be measured by standardized chemiluminescence methods. | Change from Baseline serum insulin levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum leptin levels (pg/mL) | Leptin levels will be measured by standardized chemiluminescence methods | Change from Baseline serum leptin levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum adiponectin levels (ng/mL) | Adiponectin levels will be measured by standardized chemiluminescence methods | Change from Baseline serum adiponectin levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Adiponectin/Leptin ratio (numerical ratio) | Adiponectin and leptin will be combined to report adiponectin/leptin ratio | Change from Baseline Adiponectin/Leptin ratio at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum Monocyte chemoattractant protein-1 (MCP-1) levels (pg/mL) | MCP-1 levels will be measured by standardized chemiluminescence methods | Change from Baseline serum MCP-1 levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in plasma tumor necrosis factor alpha (TNF-alpha) levels (pg/mL) | TNF-alpha levels will be measured by standardized chemiluminescence methods | Change from Baseline plasma TNF-alpha levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in plasma Interleukin 6 (IL-6) levels (pg/mL) | IL-6 levels will be measured by standardized chemiluminescence methods | Change from Baseline plasma IL-6 levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in plasma Interleukin 10 (IL-10) levels (pg/mL) | IL-10 levels will be measured by standardized chemiluminescence methods | Change from Baseline plasma IL-10 levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in plasma Intercellular Adhesion Molecule 1 (ICAM-1) levels (ng/mL) | ICAM-1 levels will be measured by standardized chemiluminescence methods | Change from Baseline plasma ICAM-1 levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in plasma Cluster of Differentiation 14 (CD14) levels (pg/mL) | CD14 levels will be measured by standardized chemiluminescence methods | Change from Baseline plasma CD14 levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in plasma oxidized low density lipoproteins (LDLox) levels (mU/L) | LDLox levels will be measured by standardized chemiluminescence methods | Change from Baseline plasma LDLox levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in serum C-Reactive protein levels (mg/L) | C-Reactive protein levels will be determined by standardized spectrophotometry methods | Change from Baseline serum C-Reactive protein levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Histidine levels in blood (umol/L) | Serum histidine levels will be determined by Liquid Chromatography coupled to tandem Mass Spectrometry | Change from Baseline Histidine levels at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Homeostatic Model Assessment from Insulin Resistance Index (HOMA-IR) | HOMA-IR will be calculated using serum glucose and insulin levels. | Change from Baseline HOMA-IR at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Fatty Liver Index (FLI) | FLI will be calculated using BMI, waist circumference, serum triglycerides and gamma glutamyl transferase levels | Change from Baseline FLI at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Triglyceride glucose index (TyG) | TyG will be calculated using serum glucose and triglycerides levels | Change from Baseline TyG at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Plasma atherogenic index | Plasma atherogenic index will be calculated as the logarithm of the TG to HDL-c ratio | Change from Baseline Plasma atherogenic index at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Dietary habits | Nutritional habits will be determined based on the results obtained from the 3-day dietary record. | Change from Baseline Dietary habits at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in Physical activity | Physical activity will be evaluated through the International Physical Activity Questionnaire (IPAQ)-short for physical activity questionnaire | Change from Baseline Physical activity at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in concomitant medication | The consumption of concomitant medication by the volunteers will be controlled by the record of concomitant medication in the case report form | Change from Baseline concomitant medication at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Change in the consumption of food supplements | The consumption of food supplements by the volunteers will be controlled by the record of food supplements in the case report form | Change from Baseline consumption of food supplements at 6 and 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Analysis of genetic polymorphisms | Single Nucleotide Polymorphisms (SNPs) in saliva samples will be analysed by Illumina sequencing | Single nucleotide polymorphisms (SNPs) in loci of genetic susceptibility to abdominal adiposity will be studied at 12 weeks after treatment for each of the two treatments (multi-ingredient and placebo) | 
| Change of Food Intake Biomarkers | Food Intake biomarkers will be evaluated in urine using Metabolomics analyses (UHPLC MS) | Change from Baseline Food Intake Biomarkers at 12 weeks for each of the two treatments (multi-ingredient and placebo) | 
| Adverse events | Possible adverse events derived from taking study's products will be recorded in the case report form | After 6 (V2) and 12 weeks (V3) of treatment period for each of the two treatments (multi-ingredient and placebo) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Nutraceuticals, Visceral Obesity, NAFLD, Metabolic risk, L-histidine, L-serine, N-Acetylcysteine, L-carnosine, Glucose metabolism, Lipid metabolism, Hepatic steatosis, Hypertension