Compositions and methods for improvement and maintenance of weight loss

Abstract

The present invention provides nutritional compositions for use in weight loss dietary interventions or in particular weight maintenance after dietary interventions, preferably after low calorie dietary interventions. The nutritional composition comprises one or more amino acids selected from the group consisting of proline, histidine, glycine and / or hydroxyproline, or a combination thereof, which can be used to assist in weight loss in obese subjects that are resistant to weight loss only by means of caloric limiting dietary intervention. In addition, the nutritional composition of the present invention may be used to help maintain the weight loss, particularly after dietary intervention.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 202080065769.5, the original application of which had an application date of September 17, 2020, and a title of “Compositions and Methods for Improvement and Maintenance of Weight Loss”. TECHNICAL FIELD

[0002] The present invention provides a nutritional composition for use in a weight loss dietary intervention or in the maintenance of weight loss, particularly after a dietary intervention, preferably after a low-calorie dietary intervention. The nutritional composition comprises one or more amino acids selected from the group consisting of proline, histidine, glycine and / or hydroxyproline, or combinations thereof, which can be used to help weight loss in obese subjects who are resistant to weight loss by means of a caloric restriction type dietary intervention alone. Furthermore, the nutritional composition of the present invention can be used to help maintain the weight loss, particularly after a dietary intervention. BACKGROUND

[0003] Obesity is a chronic metabolic disorder that has reached epidemic proportions in many parts of the world. Obesity is a major risk factor for serious co-morbidities such as type 2 diabetes, cardiovascular disease, dyslipidemia and certain types of cancer (World Health Organ Tech Rep Ser. 2000; 894:i-xii, 1-253).

[0004] It has long been recognized that low-calorie dietary interventions can be very effective in reducing body weight and that this weight loss is often accompanied by an improvement in the risk of obesity-related co-morbidities, particularly type 2 diabetes. Empirical data have shown that a weight loss of at least 10% of initial body weight leads to a significant reduction in the risk of obesity-related co-morbidities (World Health Organ Tech Rep Ser. 2000; 894:i-xii, 1-253).

[0005] However, weight loss capacity shows large inter-individual variability. Some studies show that a certain percentage of the population cannot successfully lose weight through caloric restriction diets or low-calorie diets (Ghosh, S. et al., Obesity, 2011, 19(2): 457-463). This leads to unrealistic weight loss expectations, followed by non-compliance, drop-out, and generally unsuccessful dietary interventions. In fact, studies have found that highly compliant subjects with similar BMI and undergoing the same standardized weight loss regimen show 10-fold differences in weight loss. These observations lead to the definition of an obese weight loss resistant obesity phenotype (Harper et al., Diabetes, (2002) 51:2459-2466).

[0006] Some studies suggest that subjects predicted to have a weight loss resistant obesity phenotype or a diet resistant obesity phenotype can be considered for weight loss surgery. Weight loss surgery includes various surgical procedures performed to treat obesity by altering the gastrointestinal tract to reduce nutrient intake and / or absorption.

[0007] Clearly, there is a need for more convenient and less invasive compositions and methods for weight loss resistant obese subjects or diet resistant obese subjects. SUMMARY

[0008] Nutritional compositions

[0009] In some embodiments, compositions, in particular nutritional compositions, for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention are provided. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0010] In one embodiment, the nutritional composition comprises one or more amino acids for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0011] In one embodiment, the nutritional composition comprises one or more amino acids selected from the group consisting of proline; histidine; glycine; and / or hydroxyproline; or mixtures thereof for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0012] In one embodiment, the nutritional composition comprises at least proline for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0013] In one embodiment, the nutritional composition comprises at least proline and histidine for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0014] In one embodiment, the nutritional composition comprises at least proline, histidine and glycine for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0015] In one embodiment, the nutritional composition comprises at least proline, histidine, glycine and hydroxyproline for use in weight loss during a dietary intervention or weight maintenance after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0016] In a preferred embodiment, the nutritional composition of the present application improves fat mass loss in a subject after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0017] In another preferred embodiment, the nutritional composition of the present application improves BMI in a subject after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0018] In a further preferred embodiment, the nutritional composition of the present application maintains fat mass loss in a subject after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0019] In another preferred embodiment, the nutritional composition of the present application maintains BMI in a subject after a dietary intervention. In a preferred embodiment, the dietary intervention is a low-calorie diet.

[0020] In one embodiment, the nutritional composition comprising one or more amino acids of the present application is formulated into a dietary product together with (i) a fat source, (ii) a carbohydrate source and (iii) a protein source and provided during a low-calorie dietary intervention to aid weight loss.

[0021] In one embodiment, the nutritional composition comprising one or more amino acids is administered to a subject simultaneously, sequentially or separately.

[0022] In another embodiment, the nutritional composition comprising one or more amino acids is formulated into a diet product with (i) a fat source, (ii) a carbohydrate source, and (iii) a protein source for administration together.

[0023] In one embodiment, the nutritional formulation of the diet product provides not less than 40% of the average daily caloric intake of the subject during the low-calorie meal intervention, the diet product containing the nutritional composition of the application with one or more amino acids of the application.

[0024] In one embodiment, the nutritional formulation of the diet product provides not less than 70% of the average daily caloric intake of the subject during the low-calorie meal intervention, the diet product containing the nutritional composition of the application with one or more amino acids of the application.

[0025] In one embodiment, the nutritional formulation of the diet product provides not less than 80% of the average daily caloric intake of the subject during the low-calorie meal intervention, the diet product containing the nutritional composition of the application with one or more amino acids of the application.

[0026] In one embodiment, the nutritional formulation of the diet product provides not less than 90% of the average daily caloric intake of the subject during the low-calorie meal intervention, the diet product containing the nutritional composition of the application with one or more amino acids of the application.

[0027] In one embodiment, the nutritional formulation of the diet product provides 100% of the daily caloric intake of the subject during the low-calorie meal intervention, the diet product containing the nutritional composition of the application with one or more amino acids of the application.

[0028] In one embodiment, the average daily caloric intake of the subject during the low-calorie meal intervention is from about 600 kilocalories to about 1500 kilocalories.

[0029] In another embodiment, the nutritional composition containing one or more amino acids of the application is formulated into a diet product with (i) a fat source, (ii) a carbohydrate source, and (iii) a protein source and is provided after weight loss to help maintain weight loss.

[0030] In one embodiment, the nutritional formulation of the diet product provides 40% of the daily caloric intake of the subject during the weight maintenance period after the low-calorie meal intervention, the diet product containing the nutritional composition of the application with one or more amino acids of the application.

[0031] In one embodiment, the nutritional formulation of the dietary product provides 30% of the subject's daily caloric intake during the weight maintenance period following the low-calorie meal intervention, the dietary product containing the nutritional composition of the application with one or more amino acids of the application.

[0032] In one embodiment, the nutritional formulation of the dietary product provides 20% of the subject's daily caloric intake during the weight maintenance period following the low-calorie meal intervention, the dietary product containing the nutritional composition of the application with one or more amino acids of the application.

[0033] In one embodiment, the nutritional formulation of the dietary product provides 10% of the subject's daily caloric intake during the weight maintenance period following the low-calorie meal intervention, the dietary product containing the nutritional composition of the application with one or more amino acids of the application.

[0034] In another embodiment, following the achievement of the ideal body weight, the subject can take the nutritional formulation of the dietary product during the weight maintenance period for a period of up to about 26 weeks, the dietary product containing the nutritional composition of the application with one or more amino acids of the application.

[0035] In one embodiment, the nutritional composition provides 1 g to 10 g of the subject's average daily proline intake during the meal intervention or during the weight maintenance period following the meal intervention.

[0036] In one embodiment, the nutritional composition provides 1 g to 10 g of the subject's average daily histidine intake during the meal intervention or during the weight maintenance period following the meal intervention.

[0037] In one embodiment, the nutritional composition provides 1 g to 10 g of the subject's average daily glycine intake during the meal intervention or during the weight maintenance period following the meal intervention.

[0038] In one embodiment, the nutritional composition provides 1 g to 10 g of the subject's average daily hydroxyproline intake during the meal intervention or during the weight maintenance period following the meal intervention.

[0039] In one embodiment, the subject is diagnosed with diet-resistant obesity or weight loss-resistant obesity prior to the meal intervention and is in need of the nutritional composition of the application.

[0040] In one implementation, subjects are diagnosed with diet-resistant obesity or weight-loss-resistant obesity prior to dietary intervention by measuring the levels of the following amino acids: proline, histidine, glycine, and / or hydroxyproline, in the blood, particularly in red blood cells; and when the level of any of these amino acids is below a reference value, the subject can be considered to be at risk of diet-resistant or weight-loss-resistant obesity.

[0041] Amino acids

[0042] In several embodiments, the nutritional composition of the present invention comprises one or more amino acids selected from the group consisting of proline, histidine, glycine, and / or hydroxyproline.

[0043] Methods and uses

[0044] In one embodiment, the nutritional composition of the present invention is suitable for weight loss during a low-calorie dietary intervention.

[0045] In one embodiment, the nutritional composition is suitable for improving fat loss during a low-calorie dietary intervention.

[0046] In another embodiment, the nutritional composition is suitable for improving BMI during a low-calorie dietary intervention.

[0047] In one implementation, the nutritional composition is suitable for maintaining weight loss following a dietary intervention.

[0048] In one embodiment, the nutritional composition is suitable for maintaining fat mass loss following a dietary intervention.

[0049] In another embodiment, the nutritional composition is suitable for maintaining BMI after dietary intervention.

[0050] In one embodiment, dietary intervention involves administering the nutritional composition of the present invention as part of a low-calorie diet.

[0051] In one implementation, a low-calorie diet includes reduced fat consumption.

[0052] In one implementation, a low-calorie diet includes increasing the consumption of low-fat foods.

[0053] In one embodiment, the nutritional composition and low-calorie diet of the present invention provide subjects with an average of 600 kcal to about 1500 kcal of energy per day.

[0054] In one implementation, the low-calorie diet reduces calorie intake by an average of not less than 10% per day compared to the average daily calorie intake before the low-calorie diet was administered.

[0055] In one implementation, the low-calorie diet reduces calorie intake by an average of not less than 15% per day compared to the average daily calorie intake before the low-calorie diet was administered.

[0056] In one implementation, the low-calorie diet reduces calorie intake by at least 20% per day on average compared to the average daily calorie intake before the low-calorie diet was administered.

[0057] In one implementation, the low-calorie diet reduced calorie intake compared to the average daily calorie intake during the adjacent two-week period before the low-calorie diet was administered.

[0058] In one implementation, a low-calorie diet is administered for up to 12 weeks. In another implementation, a low-calorie diet is administered for 6 to 12 weeks.

[0059] The invention also provides a method for preparing the compositions of the invention, particularly the nutritional compositions of the invention, for administering the nutritional compositions in a form suitable for subjects with weight-resistant obesity or diet-resistant obesity.

[0060] In one embodiment, the method of weight loss using the nutritional composition of the present invention is also applicable to fat loss and BMI improvement during dietary intervention.

[0061] In one embodiment, the method of weight loss using the nutritional composition of the present invention is applicable to administering a low-calorie diet to diet-resistant obesity subjects or weight loss-resistant obesity subjects.

[0062] The method of the present invention may further include determining one or more anthropometric indicators and / or lifestyle characteristics of the subject to determine whether the subject needs the composition of the present invention to achieve or maintain weight loss.

[0063] Anthropometry metrics can be selected from gender, weight, height, and age.

[0064] Lifestyle characteristics can include, for example, whether the subject is a smoker or a non-smoker, whether the subject's average alcohol consumption exceeds the recommended daily alcohol intake, or whether the subject's average exercise level is below the recommended daily exercise level.

[0065] Methods for optimizing one or more dietary interventions are also provided, including: Assess the subject’s tendency to lose weight through one or more dietary interventions and / or the subject’s tendency to maintain weight loss after one or more dietary interventions; and administer one or more dietary interventions to the subject.

[0066] In one implementation, the dietary intervention is a low-calorie diet.

[0067] In another aspect of the invention, a low-calorie diet for use in a weight loss program is provided, wherein the diet product is administered to a subject who is predicted to achieve weight loss.

[0068] In another aspect, a dietary product is provided for weight loss as part of a low-calorie diet, wherein the dietary product is administered to subjects who are predicted to achieve weight loss or maintain their weight.

[0069] In another aspect, a dietary product is provided for weight loss as part of a low-calorie diet, wherein the dietary product is administered to subjects who are predicted to achieve both weight maintenance and weight loss.

[0070] In another aspect, dietary products for use in the treatment of obesity or obesity-related diseases are provided, wherein the dietary product is administered to subjects who are predicted to achieve weight maintenance and / or weight loss.

[0071] In one embodiment, the dietary product comprises Optifast with one or more amino acids of the present invention added. ® Or Modifast ® .

[0072] Kit-of-parts

[0073] A multi-part kit for use according to the present invention is provided, the kit comprising: (i) a dietary product; (ii) a nutritional composition containing one or more amino acids of the present invention; and (iii) instructions for use thereof.

[0074] In one implementation, the dietary product comprises one or more of the following: (i) a fat source; (ii) a carbohydrate source; and (iii) a protein source.

[0075] In one implementation, the dietary product comprises two or more of (i) a fat source, (ii) a carbohydrate source, and (iii) a protein source.

[0076] In one implementation, a multipart kit comprising (i) a dietary product and (ii) a nutritional composition containing one or more of the amino acids, along with (iii) their instructions for use, is administered simultaneously, sequentially, or separately to a diet-resistant obesity subject or a weight-loss-resistant obesity subject.

[0077] In one implementation, a nutritional composition containing one or more amino acids is administered, wherein the specified one or more amino acids are administered to the subject simultaneously, sequentially, or individually.

[0078] In another embodiment, a nutritional composition containing one or more amino acids is co-formulated with a dietary product containing (i) a fat source, (ii) a carbohydrate source and (iii) a protein source for administration.

[0079] In one implementation, the kit is used during dietary interventions, particularly low-calorie dietary interventions, to improve weight loss.

[0080] In one implementation, the kit is used to maintain weight loss following dietary interventions, particularly low-calorie dietary interventions.

[0081] A method for manufacturing the multi-part sleeve of the present invention is also provided. Detailed Implementation

[0082] All percentages are by weight of the total weight of the composition unless otherwise stated. Similarly, all ratios are by weight unless otherwise stated. When pH is mentioned, the value corresponds to the pH measured using standard equipment at 25°C. As used herein, “about,” “approximately,” and “substantially” should be understood to mean a number within a range of values, such as -10% to +10% of the mentioned number, preferably -5% to +5%, more preferably -1% to +1%, and most preferably -0.1% to +0.1%.

[0083] Furthermore, all numerical ranges herein should be understood to include all integers or fractions within that range. Additionally, these numerical ranges should be understood to support claims involving any number or subset of numbers within that range. For example, disclosures of 1 to 10 should be understood to support ranges of 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, etc.

[0084] As used herein and in the appended claims, the singular form of a word includes the plural form unless the context clearly specifies otherwise. Therefore, references to “an,” “a,” and “the” generally include the plural form of the corresponding term. For example, references to “ingredient” or “method” include references to multiple such ingredients or methods. The term “and / or” used in the context of “X and / or Y” should be interpreted as “X” or “Y” or “X and Y.” Similarly, “at least one of X or Y” should be interpreted as “X” or “Y” or “both X and Y.”

[0085] Similarly, the terms “comprise,” “comprises,” and “comprising” are used to interpret them inclusively rather than exclusively. Likewise, the terms “comprise” and “or” should be considered inclusive unless the context explicitly prohibits this interpretation. However, embodiments provided in this disclosure may not contain any elements not explicitly disclosed herein. Therefore, a disclosure of one embodiment defined by the term “comprise” is also a disclosure of multiple embodiments that are “substantially composed of the disclosed components” and “composed of the disclosed components.” “Substantially composed of” means that the embodiment contains more than 50% by weight of the identified components, preferably at least 75% by weight of the identified components, more preferably at least 85% by weight of the identified components, most preferably at least 95% by weight of the identified components, for example at least 99% by weight of the identified components.

[0086] The term "example" as used herein (especially when followed by a list of terms) is for illustrative purposes only and should not be considered exclusive or comprehensive. Any embodiment disclosed herein may be combined with any other embodiment disclosed herein unless expressly indicated otherwise.

[0087] The relative terms “improved,” “enhanced,” “strengthened,” etc., refer to the effect of a composition containing one or more amino acids relative to a composition that does not contain the one or more amino acids of the present invention or has fewer of the one or more amino acids but is otherwise identical.

[0088] The terms “dietary product”, “nutritional composition”, and “nutritional formulation” mean a product or composition containing at least one amino acid of the present invention intended for consumption by an individual (such as a human) and to provide that individual with at least one nutrient. The compositions of this disclosure (including the various embodiments described herein) may comprise, consist of, or be substantially composed of the following elements: the essential elements and limitations described herein, and any other or optional ingredients, components, or limitations described herein or that may be used in a diet.

[0089] As used herein, “complete nutrition” comprises a full range of macronutrients (proteins, fats, and carbohydrates) and micronutrients in sufficient quantities to serve as the sole source of nutrition for the animal receiving the composition. From such complete nutrition compositions, an individual can obtain 100% of its nutritional needs.

[0090] "Animals" includes, but is not limited to, mammals, including but not limited to rodents; aquatic mammals; domesticated animals, such as dogs and cats; farm animals, such as sheep, pigs, cattle, and horses; and humans. When using "animals," "mammals," or their plural forms, these terms also apply to any animal capable of having a role as indicated or intended to be indicated by the context of a paragraph. As used herein, the terms "individual" or "patient" should be understood to include animals, such as mammals, and preferably humans, that receive or intend to receive treatment as defined herein. While the terms "individual" and "patient" are commonly used herein to refer to humans, this disclosure is not limited thereto.

[0091] Therefore, the terms "subject" and "patient" refer to any animal, mammal, or human who may benefit from the methods and compositions disclosed herein.

[0092] Compositions and uses thereof

[0093] Adding “compositions,” particularly “nutritional compositions” and / or “nutritional formulations,” to “dietary products” containing the amino acids of the present invention is suitable for improving weight loss during dietary interventions and maintaining weight after weight loss. Weight loss can be measured by any technique known to those skilled in the art.

[0094] As defined in this article, “weight loss” can refer to a reduction in parameters such as weight (e.g., in kilograms), body mass index (kgm-2), waist-to-hip ratio (e.g., in centimeters), body fat mass (e.g., in kilograms), hip circumference (e.g., in centimeters), or waist circumference (e.g., in centimeters).

[0095] Weight loss can be calculated by subtracting the value of one or more of the above parameters at the end of the intervention from the value of the parameters at the beginning of the intervention (e.g., according to the application of the invention).

[0096] The degree of weight loss can be expressed as a percentage change in one of the aforementioned weight phenotype parameters (e.g., the subject's weight (in kilograms) or body mass index (kgm⁻²)). For example, an individual may lose at least 10% of their initial weight, at least 8% of their initial weight, or at least 5% of their initial weight. By way of example only, a subject may lose 5% to 10% of their initial weight.

[0097] In one implementation, a weight loss of at least 10% of initial body weight results in a significant reduction in the risk of obesity-related comorbidities.

[0098] As defined in this article, “maintaining weight loss” can refer to the maintenance of parameters such as weight (e.g., in kilograms), body mass index (kgm-2), waist-to-hip ratio (e.g., in centimeters), body fat mass (e.g., in kilograms), hip circumference (e.g., in centimeters), or waist circumference (e.g., in centimeters) or body fat mass after an intervention (such as a dietary intervention).

[0099] Typically, weight loss is maintained after the period of weight loss achieved.

[0100] In one aspect, the present invention provides non-therapeutic use of the nutritional compositions of the present invention for maintaining a healthy body composition after a period of weight loss.

[0101] The degree of weight maintenance can be calculated by determining the change in one or more of the above parameters over a period of time. This period can be, for example, at least 12, 15, 20, 26, 30, 36, 40, 46, or 50 weeks.

[0102] The degree of weight maintenance can be expressed as the weight regained over a period of time after achieving weight loss, for example, as the percentage of weight lost during the weight loss period.

[0103] In one aspect, the present invention provides nutritional compositions and / or formulations to be added to dietary products of the present invention for achieving or maintaining weight loss in subjects.

[0104] The nutritional compositions and / or formulations added to the dietary products of the present invention can also improve fat loss and BMI, which can be measured by any technique known to those skilled in the art.

[0105] Individuals

[0106] The subject of this invention is preferably a mammal. In a preferred embodiment, the subject is a human. Alternatively, the individual may be a non-human mammal, including, for example, a horse, cow, sheep, or pig. In one embodiment, the individual is a companion animal, such as a dog or cat.

[0107] "Overweight" is defined as a BMI between 25 and 30 in adults. "Body Mass Index" or "BMI" is the ratio of weight (in kilograms) to the square of height (in meters). "Obesity" is a condition in which the natural energy reserves stored in the adipose tissue of animals (especially humans and other mammals) increase to a level associated with certain health conditions or increased mortality. "Obesity" is defined as a BMI greater than 30 in adults. "Normal weight" in adults is defined as a BMI between 18.5 and 25, while "underweight" is defined as a BMI less than 18.5.

[0108] Obesity-related diseases refer to any condition that an obese individual has an increased risk of developing.

[0109] Obesity-related diseases can include diabetes (e.g., type 2 diabetes), stroke, high cholesterol, cardiovascular disease, insulin resistance, coronary heart disease, metabolic syndrome, hypertension, or fatty liver.

[0110] The nutritional compositions provided for use as described herein can be used to reduce the subject's body fat mass and substantially maintain the subject's lean body mass.

[0111] "Fat mass" refers to the portion of a subject's body that is composed of fat. Fat mass can be determined using a variety of methods, such as caliper-based measurements of skinfold thickness, dual-energy X-ray absorptiometry, CT or MRI scans, or bioelectrical impedance analysis.

[0112] Reducing body fat can mean a reduction of at least 1%, at least 2%, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, or at least 50%.

[0113] "Lean body mass" or "lean body weight" refers to the body composition defined as the difference between total body weight and body fat weight. This means it includes the mass of all organs except body fat, including bones, muscles, blood, skin, and others. Maintaining lean body mass is important for optimal metabolism, normal physical activity, and overall health.

[0114] Maintaining lean body mass can basically mean that the lean body mass changes by less than 7%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1% after or during the intervention.

[0115] Preferably, most of the weight loss is due to a reduction in non-lean body mass or fat mass rather than a reduction in lean body mass.

[0116] The subjects of this invention are preferably diet-resistant obesity or weight loss-resistant obesity. “Diet-resistant obesity subjects” or “weight loss-resistant obesity subjects” can be defined as subjects who have the lowest quintile rate of weight loss (based on age and initial weight) in the subject group 6 weeks after the start of the dietary intervention (Harper et al., 2002).

[0117] Preferably, the degree of weight loss is expressed as a decrease in BMI units, where BMI decrease = ((BMI1 - BMI2)) 100) / BMI1, where BMI1 is the individual's body mass index before dietary intervention, and BMI2 is the individual's predicted body mass index after dietary intervention.

[0118] Dietary intervention

[0119] The term "dietary intervention" is understood to be an external factor applied to a subject and causing changes in the subject's diet. Preferably, the dietary intervention is a low-calorie diet.

[0120] "Low-calorie diet" includes an average calorie intake of about 600 kcal / day to about 1,500 kcal / day, more preferably about 600 kcal / day to about 1,200 kcal / day, more preferably about 700 kcal / day to about 900 kcal / day, and most preferably about 800 kcal / day.

[0121] In one implementation, the low-calorie diet may include a predetermined amount of vegetables per day, preferably up to about 400g of vegetables per day, for example, about 200g of vegetables per day. Low-fat foods may include whole wheat flour and bread, oatmeal, high-fiber breakfast cereals, whole grain rice and pasta, vegetables and fruits, dried beans and lentils, baked potatoes, dried fruit, walnuts, whitefish, herring, mackerel, sardines, salted fish, pelcherd, salmon, and lean white meat.

[0122] A low-calorie diet may include the administration of at least one “dietary product.” A “dietary product” may be formulated as a meal replacement or supplement that, for example, suppresses the subject’s appetite. Dietary products may include food products, beverage products, pet food products, food supplements, nutritional products, food additives, or nutritional formulas.

[0123] In one implementation, the dietary product may include, for example, Optifast. ® Or Modifast ® Products like these.

[0124] The "dietary products" can supplement with three servings of non-starchy vegetables, bringing the total energy intake to approximately 2.5 MJ (600 kcal / day). The diet can also be supplemented with at least 2 liters of water or other non-energy beverages daily.

[0125] In another embodiment, the "dietary product" may comprise, for example, a nutritional formulation containing the amino acid composition of the present invention, and up to at least 46.4% carbohydrates, 32.5% protein, and 20.1% fat, vitamins, minerals, and trace elements; the dietary product may be supplemented with three servings of non-starchy vegetables, resulting in a total energy intake of approximately 2.5 MJ (600 kcal / day) per meal and up to a maximum of 1500 kcal / day. The diet may also be supplemented with at least 2 liters of water or other energy-free beverages daily.

[0126] In one implementation, the low-calorie diet lasts for up to 12 weeks. Preferably, the low-calorie diet lasts for 6 to 12 weeks, more preferably 8 to 10 weeks, for example 8 weeks.

[0127] Amino acids

[0128] The term "amino acid" is a general term for organic compounds that have an amino group (-NH2) and a carboxyl group (-COOH).

[0129] In several embodiments of the present invention, specific amino acids and their derivatives are described.

[0130] Proline

[0131] In several embodiments of the present invention, "proline" or "proline derivative" is defined in nutritional compositions, methods and uses.

[0132] The amino acid "proline" is also called L-proline; L-(-)-proline; (S)-pyrrolidine-2-carboxylic acid; (2S)-pyrrolidine-2-carboxylic acid; (-)-proline, (-)-(S)-proline; proline; H-Pro-OH, 2-pyrrolidinecarboxylic acid; (S)-2-pyrrolidinecarboxylic acid; proline, (-)-2-pyrrolidinecarboxylic acid; L-pyrrolidine-2-carboxylic acid; L-α-pyrrolidinecarboxylic acid; L-proline; aminocarboxypyrrolidine.

[0133] The "proline derivative" is preferably L-proline and / or is limited to being selected from: cis-4-hydroxy-L-proline (CHP), trans-4-hydroxy-L-proline (THP), 4-hydroxy-1-methyl-proline, 1-methyl-4-anilinecarbonyloxy-proline, 1-methyl-4-anilinecarbonyloxy-proline, cis-4-hydroxymethyl-1-proline, trans-4-hydroxymethyl-D-proline, trans-4-hydroxymethyl-1-proline, trans-4-methyl-1-proline, cis-3-amino-1-proline, 1-methyl-4-phenylaminocarbonyloxy-proline-ethyl ester, 1-methyl-4-phenylaminocarbonyloxy-proline-isobutyl ester, 4-hydroxy-1-methyl-proline-ethyl ester, 4-hydroxy-1-methyl- Proline-isobutyl ester, 4-hydroxy-1-methyl-proline-ethyl ester, 4-hydroxy-proline-ethyl ester, 4-hydroxy-proline-isobutyl ester, cis-4-hydroxy-L-proline-ethyl ester, cis-4-hydroxy-L-proline-isobutyl ester, 4-hydroxy-1,1-dimethyl-proline-ethyl ester-iodine, hydroxyproline-ethyl ester, 4-hydroxy-1,1-dimethyl-proline-isobutyl ester-iodine, 4-hydroxy-1-cyclohexyl-proline-isobutyl ester, 4-hydroxy-1-diphenylmethyl-proline-isobutyl ester-hydrobromic acid, 4-hydroxy-1-methyl-proline, 4-hydroxy-1-alkyl-proline ester amides (alkyl, methyl, ethyl, propyl, pentyl, hexyl, heptyl, octyl, and nonyl), 4-hydroxy-1-diphenylmethyl-proline-isobutyl ester-hydrobromic acid. This implementation scheme covers their salts, esters, isomers, racemates, enantiomers, or prodrugs.

[0134] Histidine

[0135] In several embodiments of the present invention, "histidine" or "histidine derivative" is defined in nutritional compositions, methods and uses.

[0136] The amino acid "histidine" is also known as (S)-4-(2-amino-2-carboxyethyl)imidazolium; (S)-α-amino-1H-imidazolium-4-propionic acid; (S)-α-amino-1H-imidazolium-4-propionic acid; (S)-1H-imidazolium-4-alanine; (S)-2-amino-3-(4-imidazolyl)propionic acid; (S)-histidine; (S)-1H-imidazolium-4-alanine; 3-(1H-imidazolium-4-yl)-L-alanine; amino-1H-imidazolium-4-propionic acid ester; amino-1H-imidazolium-4-propionic acid; amino-4-imidazolium propionic acid ester; amino-4-imidazolium propionic acid; imidazolium-5-alanine.

[0137] Histidine is preferably L-histidine and / or its derivatives. L-histidine is naturally occurring and readily available from natural sources.

[0138] "Histidine derivative" is preferably selected from one or more of the following: peptides of histidine (especially dipeptides and tripeptides of histidine), peptides of histidine and one or more other amino acids (especially dipeptides and tripeptides, such as carnosine), and pharmaceutically acceptable salts of histidine.

[0139] Glycine

[0140] In several embodiments of the present invention, "glycine" or "glycine derivative" is defined in nutritional compositions, methods and uses.

[0141] The amino acid "glycine" is also known as aminoacetic acid; aminoacetic acid; aminoacetic acid; Glycocoll; Glykokoll; Glyzin; Leimzucker; 2-aminoacetic acid ester; amino-acetic acid; Glicoamin; Glycolixir; Glycosthene; Gyn-hydralin; Padil.

[0142] Glycine is preferably L-glycine and / or L-glycine ethyl ester. Non-limiting examples of suitable glycine functional derivatives include D-allylglycine; N-[bis(methylthio)methylene]glycine methyl ester; Boc-allyl-glycine-OH (dicyclohexylammonium) salt; Boc-D-cyclohexylglycine; Boc-L-cyclohexylglycine; (R)-N-Boc-(2'-chlorophenyl)glycine; Boc-L-cyclopropylglycine; Boc-L-cyclopropylglycine; (R)-N-Boc-4-fluorophenylglycine; Boc-D-propargylglycine; Amino acids; Boc-(S)-3-thienylglycine; Boc-(R)-3-thienylglycine; Da-cyclohexylglycine; La-cyclopropylglycine; N-(2-fluorophenyl)-N-(methanesulfonyl)glycine; N-(4-fluorophenyl)-N-(methanesulfonyl)glycine; Fmoc-N-(2,4-dimethoxybenzyl)-glycine; N-(2-furfuryl)glycine; La-neopentylglycine; D-propargylglycine; sarcosine; Za-phosphonoglycine trimethyl ester.

[0143] Hydroxyproline

[0144] In several embodiments of the present invention, "hydroxyproline" is defined in nutritional compositions, methods and uses.

[0145] Hydroxyproline is also known as (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid; or L-hydroxyproline. Hydroxyproline differs from proline because it contains a hydroxyl (OH) group attached to the γ-carbon atom.

[0146] Dietary products

[0147] In addition to one or more amino acids of the present invention, the dietary product may contain (i) a protein source, (ii) a fat source and (iii) a carbohydrate source.

[0148] Protein

[0149] As used herein, the term "protein" includes amino acids in their free form, molecules with between 2 and 20 amino acids (referred to herein as "peptides"), and also includes longer amino acid chains. Small peptides (i.e., chains having 2 to 10 amino acids) are suitable for this composition, either alone or in combination with other proteins. The "free form" of an amino acid is its monomeric form. Suitable amino acids include both native and non-native amino acids. The composition may comprise a mixture of one or more types of proteins, such as one or more (i) peptides, (ii) longer amino acid chains, or (iii) amino acids in their free form; and the mixture is preferably formulated to achieve a desired amino acid distribution / content.

[0150] The composition may comprise a protein providing at least a portion of one or more amino acids, and at least a portion of the protein may be derived from an animal or plant source, such as milk proteins, including one or more milk proteins, such as milk protein concentrates or milk protein isolates; caseinates or casein, such as micellar casein concentrates or micellar casein isolates; or whey proteins, such as whey protein concentrates or whey protein isolates. Alternatively or additionally, at least a portion of the protein may be a plant protein, such as one or more soy proteins or pea proteins.

[0151] Mixtures of these proteins are also suitable, for example, mixtures in which casein constitutes the majority but not all of the protein, mixtures in which whey protein constitutes the majority but not all of the protein, mixtures in which pea protein constitutes the majority but not all of the protein, and mixtures in which soy protein constitutes the majority but not all of the protein. In one embodiment, at least 10% by weight of the protein is whey protein, preferably at least 20% by weight, and more preferably at least 30% by weight. In one embodiment, at least 10% by weight of the protein is casein, preferably at least 20% by weight, and more preferably at least 30% by weight. In one embodiment, at least 10% by weight of the protein is plant protein, preferably at least 20% by weight, and more preferably at least 30% by weight.

[0152] Whey protein can be any whey protein, such as whey protein concentrate, whey protein isolate, whey protein micelles, whey protein hydrolysate, acid whey, sweet whey, modified sweet whey (of which casein-glycomacropeptide has been removed), fractions of whey protein, and any combination thereof.

[0153] Casein can be obtained from any mammal, but is preferably obtained from cow's milk, and more preferably micellar casein.

[0154] The protein can be unhydrolyzed, partially hydrolyzed (i.e., peptides having a molecular weight of 3 kDa to 10 kDa and an average molecular weight of less than 5 kDa), or fully hydrolyzed (i.e., where 90% of the peptides have a molecular weight of less than 3 kDa), for example, in the range of 5% to 95% hydrolysis. In some embodiments, the peptide distribution of the hydrolyzed protein can be in different molecular weight ranges. For example, the majority of the peptides (> 50 molar percentage or > 50 wt%) may have a molecular weight in the range of 1 kDa to 5 kDa, 5 kDa to 10 kDa, or 10 kDa to 20 kDa.

[0155] Fat

[0156] In one embodiment, the composition comprises a fat source. The fat source may include any suitable fat or mixture of fats. Non-limiting examples of suitable fat sources include vegetable fats (such as olive oil, corn oil, sunflower oil, high-oleic sunflower oil, rapeseed oil, low-erucic acid rapeseed oil, hazelnut oil, soybean oil, palm oil, coconut oil, blackcurrant seed oil, borage oil, lecithin, etc.), animal fats (such as milk fat); or combinations thereof.

[0157] Carbohydrate

[0158] In one embodiment, the composition comprises a carbohydrate source. Any suitable carbohydrate may be used in the composition, including but not limited to starch (e.g., modified starch, amylose, tapioca starch, corn starch), sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, xylitol, sorbitol, or combinations thereof.

[0159] The carbohydrate source is preferably no more than 50% of the energy of the composition, more preferably no more than 36% of the energy of the composition, and most preferably no more than 30% of the energy of the composition.

[0160] Nutritional formulation

[0161] The composition can be any kind of composition formulated to be suitable for human and / or animal consumption. For example, the composition may be selected from food compositions, dietary supplements, nutritional compositions, nutritional medicines, powdered nutritional products reconstituted with water or milk before consumption, food additives, pharmaceuticals, beverages, and drinks. In one embodiment, the composition is an oral nutritional supplement (ONS), a complete nutritional formula, a medicine, a medical product, or a food product. In a preferred embodiment, the composition is administered to an individual in the form of a beverage. The composition may be stored in powder form in sachets and then suspended in a liquid such as water for use.

[0162] In some embodiments, the composition is administered to an individual in a single dosage form, i.e., all compounds are present in one product, which is provided to the individual in combination with a dietary supplement. In other embodiments, the composition is administered co-administered in separate dosage forms, e.g., at least one component is separate from one or more other components of the composition.

[0163] Examples

[0164] The following non-limiting embodiments present scientific data supporting the concept of developing and administering compositions comprising one or more amino acids of the present invention, wherein the composition further comprises an effective amount of one or more amino acids such that the effective amount allows the composition to be used for weight loss needs or for weight maintenance after weight loss. It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. These changes and modifications may be made without departing from the spirit and scope of the subject matter of the invention and without diminishing its intended advantages. Therefore, such changes and modifications are intended to be covered by the appended claims.

[0165] Example 1: Grouping of meal replacement OPTIFAST dietary intervention

[0166] The clinical trial OPTIWIN tested the effectiveness of the OPTIFAST regimen (OP) for weight loss compared to a food-based (FB) dietary plan, which is a full meal replacement dietary intervention (Ard et al. Obesity (SilverSpring). 2019 Jan; 27(1): 22-29).

[0167] OPTIWIN is an open-label, multicenter, randomized, controlled clinical trial. Nine participating U.S. centers include five private clinics and four academic medical centers. Participants must be between 18 and 70 years of age and have a BMI of 30 kg / m². 2 Up to 55kg / m 2Non-smokers. Key exclusion criteria include recent weight loss or use of weight loss medications, previous weight loss surgery, organ failure, type 1 diabetes, or hemoglobin A1c. 1c The study was conducted in a pool of participants with a >10% chance of being hospitalized for cardiovascular disease or mental health issues within the past 6 months, alcohol or drug dependence, a positive screening for potential eating disorders, or poorly controlled depression. Volunteers were initially screened by telephone to assess eligibility, followed by in-person visits to confirm eligibility and obtain written informed consent. Eligible individuals completed a 7-day adaptation diet, which included one serving of a meal replacement (OPTIFAST; Nestlé Health Science, Bridgewater, New Jersey) daily to assess product tolerance. A final screening visit was conducted after the adaptation period to assess adherence to the adaptation diet, self-monitoring tasks, and final eligibility. Following review by the ethics review committees at each site, the study was supervised and approved by the central ethics review committee (quorum) and registered on ClinicalTrials.gov as NCT02635698.

[0168] Treatment groups

[0169] Participants were randomly assigned to either the OPTIFAST (OP) group or the food-based (FB) group.

[0170] Optifast group (OP) : Participants with a BMI < 45 were instructed to use five meal replacements (MRs), consisting of a daily Optifast diet (800 kcal) comprising 40% protein, 40% carbohydrates, and 20% fat. Participants with a BMI of 45 to 49.9 received six MRs daily (960 kcal); participants with a BMI ≥ 50 received six MRs plus one lean protein meal (3 to 4 ounces) and one serving of non-starchy vegetables daily (1,100 to 1,200 kcal). Participants adhered to their prescription for 12 to 16 weeks, as determined by the provider and according to patient preference, before gradually reintroducing food until week 26. After week 26, participants gradually increased their calorie intake to achieve weight stabilization. During this period, participants were advised to use MRs once or twice daily to promote weight loss maintenance.

[0171] Participants regularly visited their clinicians to address health issues and adjust medications, completing a total of 11 medical monitoring visits during the first 26 weeks and 4 more between weeks 27 and 52. Laboratory data (including basal metabolic or comprehensive metabolic studies) for medical monitoring were obtained at weeks 2, 4, 6, 8, 10, 12, and 16. Participants also had individual consultations with trained interventional personnel, completing 16 individual consultation visits during the first 26 weeks and 11 more between weeks 27 and 52.

[0172] Food-based group (FB) : During effective weight loss (weeks 0 to 26), FB participants were prescribed a calorie-restricted diet emphasizing low fat intake (25% to 30% of total calories). They were advised to reduce their calorie intake by 500 to 750 kcal less than their estimated total energy expenditure (calculated based on resting metabolic rate measured via indirect calorimetry (MedGem; MicrolifeUSA, Inc., Clearwater, Florida) plus an activity coefficient based on self-reported physical activity). To balance the incentive of free MR in the OP group, FB participants received a total $800 in gift certificates were given to offset food costs during the effective weight loss phase. By week 26, they had also completed two medical monitoring visits and seven individual counseling visits. During the maintenance phase (weeks 27 to 52), they completed two medical monitoring visits and five individual counseling visits.

[0173] Analysis

[0174] The sample size was calculated primarily to demonstrate the effectiveness of OP compared to FB in terms of percentage weight change at 52 weeks. To achieve 90% statistical power based on a two-sided test with the hypothesis α=0.05, 113 participants per study group were needed to detect the mean difference of 5% weight change, with an estimated standard deviation (SD) of 11.5%. Due to an estimated loss to follow-up rate of 30%, we planned to recruit 300 participants to provide 150 participants per study group.

[0175] The primary outcome of the synergistic analysis was the percentage change in body weight from baseline to weeks 26 and 52, measured using a graded process. We initially planned to test the week 26 results. If this analysis was statistically significant ( P If the significance level is < 0.05, then the week 52 endpoint will be tested at a significance level of α = 0.05. However, if the week 26 analysis is not statistically significant, then the week 52 analysis will be exploratory rather than deterministic.

[0176] The primary analysis cohort was the modified intention-to-treat (mITT) cohort, consisting of randomized subjects known to have initiated the study program and received at least one post-baseline weight assessment. For the primary analysis, we used a repeated measures model with random and fixed effects to compare percentage weight change between the OP and FB groups. The linear mixed model included the random intercept for subjects, fixed visit effects, fixed treatment effects, and fixed baseline weight effects, as well as visit-treatment interaction terms. The model also included age, race, sex, site, and diabetes status as covariates. The final model was fitted using the SAS PROCMIXED procedure (using RANDOM and LSMEANS statements) (SAS Institute, Inc., Cary, North Carolina). Sensitivity analyses employed multiple imputation (MI) for missing weight values ​​at the planned post-baseline visits (weeks 12, 26, 40, and 52) in the mITT cohort. Imputation was performed separately for each treatment group using the SAS PROC MI procedure (imputation = 50). The primary outcome was then computed from the imputed dataset. For secondary outcomes including body composition and frequency of adverse events (AEs), standard procedures were used. t Tests comparing continuous measurements using the χ² method. 2 Test and compare category measurements.

[0177] Results

[0178] We screened 463 participants and confirmed 330 participants as eligible (164 were randomly assigned to the OP and 166 to the FB). Of the 57 participants, some declined further participation after learning of their randomization assignments (9 OP; 16 FB), and others discontinued participation without weight tracking (20 OP; 12 FB). These participants were demographically similar to the mITT group, except that they were younger (42.5 years). P = 0.006) and reported less weight loss on average (0.1 kg) after previous attempts; P< 0.001). The remaining 273 participants (135 OP; 138 FB) constituted the mITT study cohort (82.7% of the randomized participants). Within the mITT study cohort, 19 participants (14.1%) in the OP group and 25 participants (18.1%) in the FB group discontinued participation. Participants were typically middle-aged (47.1 ± 11.2 years), predominantly female (82%) and white (71%) (Table 1). 52% had prediabetes or diabetes, 33% were receiving treatment for hypertension, and 21% were receiving treatment for dyslipidemia. OP and FB participants attended 53.5% and 46.2% of the weekly group behavior meetings, respectively.

[0179] Table 1

[0180] Summary of demographic and baseline characteristics by treatment group

[0181] Use of standards t Tests were used to compare continuous variables. Except for race, categorical variables were compared using the χ² test; Fisher's exact test was used for race due to small cell counts.

[0182] a Drug use includes use before the study and during the study, or use that begins during the study and continues throughout the study or ends before the study is completed.

[0183] Weight loss results

[0184] Both treatment groups achieved weight loss at weeks 26 and 52 (for OP, the percentage weight loss at week 26 was 12.4% ± 0.6%, while for FB it was 6.0% ± 0.6% (difference 6.4% ± 0.9%)). P < 0.001). At week 52, the percentage weight loss was 10.5% ± 0.6% for OP and 5.5% ± 0.6% for FB (difference 4.9% ± 0.9%). P < 0.001). Sensitivity analysis showed similar estimates for percentage weight loss. For OP, using MI to handle missing data showed weight loss of 12.1% ± 0.6% and 10.1% ± 0.7% at weeks 26 and 52, respectively. In contrast, weight loss for FB was 6.0% ± 0.6% and 5.5% ± 0.7% at weeks 26 and 52, respectively. All differences between OP and FB were within the range of 0.001. P It remains significant at < 0.001.

[0185] Relative weight change in the treatment group at weeks 26 and 52. Percentage weight change was calculated using least squares from a linear mixture model. The linear mixture model included random intercept subject effects, fixed visit effects, fixed treatment effects, fixed baseline weight effects, visit-treatment interaction terms, and covariates (including age, race, sex, site, and reported baseline diabetes status).

[0186] At week 52, a higher proportion of OP participants achieved 5% weight loss (63.7%) and 10% weight loss (43.7%) compared to FB (42% and 21.7%, respectively). P < 0.001). The percentage of people in the OP group who lost 15% or more of their initial weight was more than double that in the FB group (30% vs. 12%). P < 0.001). We also examined the proportion of participants who were unresponsive to their assigned treatment, defined as those who failed to lose ≥ 3% of their initial body weight. At week 26, 15.6% of OP participants and 39.1% of FB participants were unresponsive. By week 52, 23.7% of OP participants and 43.5% of FB participants were unresponsive.

[0187] Changes in body composition

[0188] Table 2 shows the changes in body composition outcomes. Changes in waist circumference occurred concurrently with weight loss in both treatment groups; however, at weeks 26 and 52, the reduction was greater in the OP group compared to the FB group. Changes in body composition, measured by dual-energy X-ray absorptiometry, followed a similar pattern. By week 52, for OP participants, the mean reduction in total fat mass was 9.7 ± 10.4 kg. For FB participants, the mean reduction in total fat mass at week 52 was 3.5 ± 6.6 kg. A reduction in lean body mass was also observed in both treatment groups. The absolute difference was statistically significant; however, the proportion of weight loss relative to lean body mass at week 52 was similar (23% for OP; 25.5% for FB). There were no differences in overall bone mineral density changes between the treatment groups.

[0189] Table 2: Changes in body composition results from baseline

[0190] The change relative to the baseline is defined as the post-baseline value - the baseline value. (Using standard...) t The test compares the differences.

[0191] Participants in the OP treatment group lost 10.5% of their initial weight by week 52, representing nearly double the weight loss observed in FB. A larger proportion of participants in OP lost 5%, 10%, or 15% of their weight, and a greater percentage reported at least 3% weight loss. OP participants exhibited greater fat mass loss, and OP was well-tolerated and safe. Overall, OP proved to be a more effective treatment than FB.

[0192] The OP group achieved greater weight loss and experienced less weight rebound after 12 months. Continued use of meal replacements (such as OPIFAST) for weight maintenance after weight loss may be beneficial due to the characteristics of MR associated with lower energy intake (e.g., portion control, low calories, reduced choices) and changes in individual subjective food cravings, objectively supported by functional magnetic resonance imaging (fMRI) showing contemporaneous changes in brain regions responsible for regulating food intake and reward. Continued treatment and use of meal replacements may be important for sustained weight loss.

[0193] Example 2: Method for measuring amino acids

[0194] Plasma samples from subjects in the above-mentioned OP group were analyzed at baseline, week 26, and week 52 to determine amino acid concentrations. Specifically, proline, histidine, glycine, and hydroxyproline were measured.

[0195] Amino acid analysis was performed using tandem mass spectrometry (UPLC-MS / MS). Separation and analysis were performed on an Accela UHPLC 1250 pump (Thermo Fisher Scientific Inc., Waltham, MA, USA) connected to a TSQ Quantum Vantage triple quadrupole (Thermo Fisher Scientific Inc., Waltham, MA, USA), equipped with a heated electrospray ionization (H-ESI) source. Chromatographic separation was achieved using gradient elution on a reversed-phase UPLC XSelect HSST3 2.5µm, 100 × 2.1mm inner diameter column (Waters Corporation, Milford, MA, USA). The injection volume was 10µL, and the total run time for the analysis was 13 minutes. The plasma sample was thawed and vortexed for 10 s. 50µL of plasma was transferred to a 1.5mL microcentrifuge tube. 10µL of IS solution was added to the plasma. Then, 50 μL of TCEP solution followed by 140 μL of methanol + 1% FA was added to the mixture. The tube was placed in a multi-tube vortex mixer and vortexed at 1350 rpm for 15 minutes at 4 °C, followed by centrifugation at 14,500 rpm for 5 minutes. The supernatant was pipetted, filtered through a 0.22 µm filter, and placed in a vial for LC-MS / MS analysis.

[0196] The sparse partial least squares (sPLS) method was used to test the correlation between changes in blood amino acid during weight loss and changes in anthropometric variables throughout the intervention period (i.e., after WM). The sample consisted of 71 participants with data at baseline (V1), 12 weeks later (V2), and 24 weeks later (V3) of the weight loss protocol.

[0197] The Sparse Least Squares (sPLS) method implemented in the mixOmics R package combines integration and variable selection on two datasets (amino acids and anthropometric variables) in a one-step strategy. This method is a multivariate approach involving two data matrices X (e.g., anthropometrics) and Y (amino acids). sPLS outperforms traditional multiple regression by modeling the structure of the two matrices. Unlike traditional multiple regression models, sPLS is not limited to uncorrelated variables. Among the many advantages of sPLS is its ability to handle many noisy, collinear (correlated), and missing variables, and it can also model several response variables Y simultaneously. Variable selection is achieved by introducing a LASSO penalty onto a pair of loading vectors. A Cluster Image Map (CIM) is provided to summarize and visualize the sPLS results. Missing data is imputed by bagging. This method is suitable for bagged tree models for each predictor (as a function of all other factors). CIM is based on hierarchical clustering applied simultaneously to the rows and columns of the real-valued similarity matrix obtained via the sPLS results. A network graph is also provided to represent the correlation between the two sets of variables. The network graph is inferred from the paired association scores between variables X and Y provided by sPLS. The threshold only represents variables X and Y with association scores greater than a given cutoff point.

[0198] The analysis assumes the use of a 3-component model for visualization and interpretation. The results are reported in Table 3 according to the CIM element values.

[0199] The analysis highlighted a strong correlation between changes in blood amino acids during weight loss (V2 / V1) and the body composition achieved at the end of the weight maintenance period (V3 / V1). Specifically, changes in proline were positively correlated with changes in lean body mass and negatively correlated with changes in BMI and body fat mass. Furthermore, changes in glycine and histidine were negatively correlated with changes in body fat mass and BMI. These observations are interpreted by the fact that (i) subjects exhibiting no or weak loss of proline, histidine, and glycine achieved greater loss of body fat mass and BMI, and (ii) greater lean body mass maintenance was observed in subjects exhibiting weak loss of proline.

[0200] Hydroxyproline is produced by the hydroxylation of the amino acid proline. Hydroxyproline content is a well-known marker of collagen catabolism, particularly bone resorption or tissue degradation (including muscle damage). Changes in the hydroxyproline-to-proline ratio are negatively correlated with changes in lean body mass and positively correlated with changes in body fat mass and BMI. Specifically, a higher conversion rate of hydroxyproline to proline results in greater lean body mass loss.

[0201] Table 3: CIM values

Claims

1. A nutritional composition for use in weight loss, wherein the nutritional composition comprises one or more amino acids selected from the group consisting of proline, histidine, glycine and / or hydroxyproline or mixtures thereof.

2. A nutritional composition for use in weight maintenance, wherein the nutritional composition comprises one or more amino acids selected from the group consisting of proline, histidine, glycine and / or hydroxyproline or mixtures thereof.

3. The nutritional composition for use in weight loss during a low calorie meal intervention or weight maintenance after a low calorie meal intervention according to claim 1 or 2, wherein the nutritional composition comprises one or more amino acids selected from the group consisting of proline, histidine, glycine and / or hydroxyproline or mixtures thereof.

4. The nutritional composition for use according to claim 3, wherein the low calorie meal intervention is between 600 and 1500 kcal / day for up to 12 weeks for adults.

5. The nutritional composition for use according to any one of claims 1 to 4, wherein the nutritional composition comprises at least proline.

6. The nutritional composition for use according to any one of claims 1 to 4, wherein the nutritional composition comprises at least proline and histidine.

7. The nutritional composition for use according to any one of claims 1 to 4, wherein the nutritional composition comprises at least proline, histidine and glycine.

8. The nutritional composition for use according to any one of claims 1 to 4, wherein the nutritional composition comprises at least proline, histidine, glycine and hydroxyproline.

9. The nutritional composition according to any one of claims 1 to 8, wherein the nutritional composition improves fat mass loss in a subject after a low calorie meal intervention.

10. The nutritional composition according to any one of claims 1 to 8, wherein the nutritional composition improves BMI in the subject after a low calorie meal intervention.

11. The nutritional composition according to any one of claims 1 to 8, wherein the nutritional composition maintains fat mass loss in the subject after a low calorie meal intervention.

12. The nutritional composition according to any one of claims 1 to 8, wherein the nutritional composition maintains BMI in the subject after a low calorie meal intervention.

13. The nutritional composition for use according to any one of claims 1 to 12, wherein the one or more amino acids are administered simultaneously, sequentially or separately to a diet resistant obese subject or a weight loss resistant obese subject.

14. The nutritional composition for use according to any one of claims 1 to 13, wherein the one or more amino acids are administered simultaneously to a diet resistant obese subject or a weight loss resistant obese subject in a diet product together with the low calorie meal intervention.

15. A kit-of-parts comprising (i) a dietary product and / or (ii) a nutritional composition comprising one or more amino acids according to any one of claims 1 to 14 and (iii) instructions for their simultaneous, sequential or separate administration to a diet-resistant obese subject or a weight loss-resistant obese subject.

16. A method of reducing the BMI of a subject in need thereof, the method comprising administering to the subject in need thereof a nutritional composition comprising one or more amino acids according to any one of claims 1 to 14.

17. A method of maintaining the BMI of a subject after weight loss, the method comprising administering to a subject in need thereof a nutritional composition comprising one or more amino acids according to any one of claims 1 to 14.

18. A method of reducing the fat mass of a subject in need thereof, the method comprising administering to the subject in need thereof a nutritional composition comprising one or more amino acids according to any one of claims 1 to 14.

19. A method of maintaining the fat mass of a subject after weight loss, the method comprising administering to a subject in need thereof a nutritional composition comprising one or more amino acids according to any one of claims 1 to 14.

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