MXPA06001190A - Use of pinolenic acid - Google Patents

Abstract

Method of managing weight by administering a composition including pinolenic acid or derivative such as a glyceride thereof. The composition may be pine nut oil or a derivative thereof and may be used as a food supplement or food product such as margarine or as a pharmaceutical composition, e.g. in tablet form. The composition may also include other components such as linoleic acid or other fatty acid or derivative thereof.

Description

USE OF PINOLENIC ACID DESCRIPTIVE MEMORY This invention relates to compositions that can be used for weight control, to control the food ration and appetite, and to control and / or reduce body weight. The compositions contain pinolenic acid, or a derivative thereof. Obesity is becoming more and more frequent in individuals of developed societies. In general, a condition of overweight and obesity results from an imbalance in the contribution and use of energy. The cause of the energy imbalance for each individual may be due to a combination of several factors, and depends on a myriad of genetic and environmental determinants. Obesity may be a contributing factor to the increased incidence of several diseases including coronary artery disease, hypertension, stroke, diabetes and certain cancers. Weight reduction is often recommended as the first course of action for patients suffering from these diseases related to obesity. In an attempt to control the total body weight, an individual can carry out weight control, whose objectives may differ, depending on the needs of the individual. For example, while obese or overweight individuals may wish to lose body weight, other individuals may wish to maintain a body weight at a substantially constant level. Even once a person has lost body weight, weight control is often required to prevent the person from regaining the previously lost body weight. The most effective weight loss programs typically include a reduced calorie diet and / or increased energy expenditure. Over time, many people who carry out weight control experience increased levels of hunger and / or cravings for high-sugar foods. This can lead to individuals moving away from their weight control regimen. Therefore, there is a need to develop new and effective ways to support weight control, and to help individuals continue with their weight management regimen. It is an object of the present invention to provide new means to provide this support. The pinolenic acid (ie, fatty acid 5, 9, 12 C18: 3, a fatty acid with 18 carbon atoms having three double bonds at positions 5, 9 and 12) is present, for example, in walnut oil of pine and fractions thereof (J Am OH Chem Soc 1998, 75, pp. 45-50). It is known that pine nut oil, and in this way pinolenic acid, can be applied in food products; see, for example, FR-A-2756465, which describes the use of a concentrate with 15% pinolenic acid in food additives. It is described that the presence of pinolenic acid provides a hypolipidemic effect to the composition. WO 98/43513 discloses that nail files can be coated with pinolenic acid, and that this inhibits the occurrence of infections after the use of the files. According to JP-A-61238729, pine nut oil can be used as an anti-cholesteric agent. Other documents in which effects of pinologenic acid on health are described, include JP-A-61058536, which describes a very generic activity beneficial for human health. This activity for health is not described in more detail. Sugano, Brit J. of Nutr 72 (1994) 775-783, describes many health effects of diets containing pinolenic acid. The aforementioned health effects are hypocholesterolemic effects, effects on platelet aggregation induced by ADP, on the production of aortic prostacyclin and on blood pressure. EP-A-1088552 describes the use of pinolenic acid as an anti-inflammatory agent. Matsuo et al., Prostaglandins, Leukotrienes and Essential Fatty Acids (1996), 55 (4), 223-229, describe the effects of gamma-linolenic acid and pinolenic acid on the immune parameters of Brown-Norway rats. Nevertheless, none of the prior art documents indicates that pinolenic acid or a derivative thereof could be used to treat or prevent obesity and / or for weight control. US 6,429,190, US 2002/0119948 and US 2002/0119915 disclose compositions for extending satiety, comprising a source of calcium, protein and a C 12 to C 18 fatty acid. Oleic acid is the fatty acid described in the documents. WO 02/088159 describes pharmaceutically active uridine esters, and combinations comprising their uridine compound and acid constituent, and their use in a wide variety of medical applications. There is no description of the use of free fatty acids alone, nor is any example given for the treatment of obesity. EP-A-1504778, published on February 9, 2005, describes an implantable pump for the treatment of obesity. The pump may comprise a fatty acid, but no description of pinolenic acid exists. CN-A-1377673 refers to the use of a pine nut oil to treat cerebral and cardiac vascular diseases and adiposity caused by hyperlipidemia, as well as diabetes caused by hyperglycemia. The present invention provides the use of pinolenic acid or a derivative thereof in the manufacture of a composition for weight control, reducing the feeling of hunger and / or increasing satiety. The invention also provides the use of pinolenic acid or a derivative thereof to reduce the sensation of hunger and / or increase satiety. In this manner, the compositions of the invention are suitable for treating or preventing obesity, and for weight control, and comprise pinolenic acid or a derivative thereof. Pinollenic acid derivatives that can be used in the present invention include salts of pinolenic acid and alkyl esters.

Other derivatives of pinolenic acid which can be used in the invention are isomers of pinolenic acid such as, for example, geometric isomers (having one or more double trans bonds, the double bonds in the pinolenic acid are all cis) and / or compounds having 18 carbon atoms and three double bonds with one or more of the three double bonds at a different position in the alkyl chain compared to the pinolenic acid including, for example, gamma-linolenic acid, alpha-linolenic acid, acid punic acid, eleseaaric acid, and its alkyl salts and esters. The invention also encompasses, therefore, the use of alpha-linolenic acid or a derivative thereof for weight control, reducing the feeling of hunger and / or increasing satiety; the use of gamma-linolenic acid or a derivative thereof for weight control, reducing the feeling of hunger and / or increasing satiety; the use of punic acid or a derivative thereof for weight control, reducing the sensation of hunger and / or increasing satiety; and the use of eleseaárico acid (also denominated alfa-elefesárico acid), or a derivative of same, for control of weight, reducing the sensation of hunger and / or increasing the satiety. The invention also contemplates pinolenic acid or a derivative thereof for treating or preventing obesity, and / or for weight control. The pinolenic acid or derivative thereof can be used alone, or it can be a component of a composition comprising other edible and / or pharmaceutically acceptable components. Preferably, the composition further comprises 10 to 60% by weight of linoleic acid and 5 to 52% by weight of oleic acid, such as free acids or glycerides (for example, mono-, di- or tri-glycerides). In addition or alternatively, the composition may comprise from 0.5 to 5% by weight of taxoleic acid. It has surprisingly been found that the pine nuts oil fatty acids, and thus pinolenic acid, can be used to stimulate the release of cholecystokinin (CCK). CCK is a peptide released after the consumption of food. When food is consumed, the CCK-releasing protein (CCKRP) is released into the small intestine. CCKRP stimulates the release of CCK from intestinal cells. The release of CCK generates behavioral symptoms associated with satiety. In addition, increased CCK levels can increase the levels of IgA in the intestine that can increase mucosal immunity. Increased levels of IgA in the intestinal tract may offer increased protection against invading microorganisms. Therefore, the invention also contemplates a method for enhancing immunity (eg, mucosal immunity), comprising the administration of pinolenic acid or a derivative thereof, pinolenic acid or a derivative thereof to increase immunity (e.g. mucosal immunity), and the use of pinolenic acid or a derivative thereof, in the manufacture of a composition for enhancing immunity (eg, mucosal immunity). Increased immunity in the treatment and / or prevention of infections, such as bacterial or viral infections, can be used.

In broad terms, the invention relates to the use of straight chain carboxylic acids (and their derivatives, such as salts and esters) having 18 carbon atoms and three double bonds for weight control and / or to treat or prevent obesity. Weight control can include reducing the feeling of hunger and / or increasing satiety. The compositions of the invention may be in any suitable form such as a food supplement, a pharmaceutical composition or a food composition. The term "composition" means that the pinolenic acid or derivative thereof may be present with one or more other components (for example, as they are present in pine nut oil). The one or more other components may be present in admixture with the pinolenic acid or derivative thereof, or they may be part of the product package (eg, the capsule in which the pinolenic acid or derivative thereof is encapsulated). The compositions of the invention typically comprise of 0. 3 to 100% by weight, preferably 5 to 80% by weight, more preferably 10 to 50% by weight (such as 10 to 30% by weight or 20 to 30% by weight) of pinolenic acid or a derived from it. The person skilled in the art will appreciate that the percentage of pinolenic acid or a derivative thereof in a composition of the invention will depend on the nature of the composition. For example, it is likely that the pinolenic acid or derivative thereof represents a greater percentage of the total weight of a pharmaceutical composition or a food supplement, than a food composition.

When the composition of the invention is in the form of a food supplement or a pharmaceutical product, the pinolenic acid or derivative thereof or a mixture containing one of these compounds, may be in encapsulated form in an encapsulating material of adequate quality for food . Suitable encapsulating materials are well known in the art and include, for example, gelatin and glycerol. The pinolenic acid used in the present invention may be in the form of a free fatty acid, or a pinolenic acid derivative or mixtures thereof, including mixtures of different derivatives. The derivatives are non-toxic and edible and preferably include, for example, salts and esters. Other pinolenic acid derivatives which can be used in the invention are pinolenic acid isomers such as, for example, geometric isomers (having one or more trans double bonds, the double bonds in the pinolenic acid are all cis), and / or compounds having 18 carbon atoms and three double bonds with one or more of the three double bonds at a different position in the alkyl chain compared to the pinolenic acid including, for example, gamma-linolenic acid, alpha-linolenic acid, punic acid, eleseaaric acid, and its alkyl salts and esters. Suitable salts include salts with cations of suitable quality for food, such as sodium salts. Suitable esters include alkyl esters having from 1 to 6 carbon atoms. Preferred esters are mono-, di- and tri-glycerides. Preferably, the compositions of the invention are free from (or substantially free of, ie, containing less than 0.1 mg of) uridine and / or nucleoside and / or nucleotide esters selected from the group consisting of uridine, deoxyuridine, monophosphate uridine, deoxyuridine monophosphate, and / or pharmaceutically acceptable salts thereof. A suitable source for the pinolenic acid used in the present invention is pine nut oil, or concentrates thereof. For example, pinolenic acid glycerides can be obtained from pine nut oil, or concentrates thereof. Preferably, an oil or concentrate with a pinolenic acid content or a derivative thereof, of more than 15% by weight or more than 28% by weight is used. The compositions of the invention may comprise one or more other fatty acids (ie, straight chain carboxylic acids having from 12 to 24 carbon atoms). Examples of other fatty acids suitable for use in the present invention include linoleic acid, oleic acid, taxoleic acid, juniperonic acid, sciadonic acid, saturated fatty acids, conjugated linoleic acid (optionally as a mixture of enriched isomers) and EPA (eicosapentanoic ) and DHA (docosahexanoic) (optionally as a mixture of enriched isomers of EPA or DHA). Enrichment includes alteration of the mixture of isomers normally present (eg, in a natural product), such as an alteration in the relative amounts of different geometric isomers. In these compositions, the other fatty acid or each of the other fatty acids can be present independently as a free fatty acid or as a derivative thereof (including a mono-, di- or tri-glyceride and salts thereof), or as a mixture of it. The pinolenic acid or derivative thereof is optionally mixed with these additional fatty acids or glycerides, before they are used to prepare a composition in accordance with the present invention. When the compositions of the invention contain one or more fatty acids and / or glycerides in addition to the pinolenic acid or derivative thereof, the additional fatty acids and / or glycerides are preferably selected from liquid oils, such as soybean oil, sunflower oil , rapeseed oil and cottonseed oil; cocoa butter and cocoa butter equivalents; palm oil and fractions thereof; enzymatically obtained fats; fish oils and fractions thereof; conjugated linoleic acid and mixtures of enriched isomers; gamma-linolenic acid and enriched mixtures thereof; hardened liquid oils; and mixtures thereof. Mixtures containing one or more additional fatty acids or glycerides preferably exhibit solid fat contents measured by NMR pulse over unstabilized fats of: N20 = 1-80, preferably 5-45, N35 less than 20, preferably less than 10, more preferably less than 5. These values were obtained by melting the fat at 80 ° C, cooling to 0 ° C and keeping the fat at 0 ° C for 30 minutes, after which the fat was heated to the temperature of measurement N, and kept at that temperature for 30 minutes before measuring the value of N.

Fatty acid mixtures which are used to produce the compositions of the invention, preferably comprise from 1.5 to 60% by weight, more preferably from 28 to 60% by weight of pinolenic acid, from 10 to 60% by weight of linoleic acid, and from 5 to 52% by weight of oleic acid, for example, from 25 to 85% by weight (linoleic acid plus oleic acid), from 0 to 70% by weight, for example, from 25 to 65% by weight (acid trans fat plus saturated fatty acid). The content of trans fatty acid is preferably less than 10% by weight. An example of a suitable mixture is one in which the content of trans fatty acid plus saturated fatty acid is less than 10% by weight. Alternatively, the compositions of the invention may be free or substantially free of fatty acids other than pinoleic acid (i.e., they may contain less than 1% by weight, more preferably less than 0.1% by weight, such as less than 0.01% by weight of other fatty acids of C12 to C24). The compositions of the invention may comprise calcium and / or magnesium sources. In addition or alternatively, the compositions may comprise protein (including protein hydrolysates). The combination of pinolenic acid and calcium and / or magnesium and / or protein in the compositions of the present invention can increase the release of CCK from intestinal cells. In another embodiment, the compositions of the invention may be free or substantially free of calcium and / or protein ions (ie, each of these components is present in the compositions in an amount of less than 1% by weight, more preferably less than 0.1% by weight, such as less than 0.01% by weight). Typically, the compositions of the invention contain pinolenic acid and / or a derivative thereof as the sole active component. The compositions of the present invention can be used to help control body weight, for example, to help maintain body weight at a substantially constant level, or to help reduce body weight. In other words, the use of the compositions can help to thin the body, for example, helping to induce fat loss. The use of the compositions of the invention can help reduce the intake of calories. This can help maintain the body weight at a substantially constant level, and / or can help reduce body weight and / or help lose weight. The reduction in body weight can increase energy levels. The use of the compositions of the invention can reduce the feeling of hunger and / or increase satiety and / or reduce the desire for high calorie foods, for example, by allowing less room in the stomach for high calorie foods . In particular, the use of the compositions of the invention can enhance and / or extend satiety or fullness before, during and / or after a food. The use of the compositions of the invention can reduce the feeling of hunger during the diet regime, and therefore increase the success rate of a diet. The consumption of the composition of the invention can stimulate the release of CCK and / or IgA. The use of the compositions of the invention can help reduce appetite, for example, by increasing satiety and a feeling of fullness and / or fullness. The present invention also provides a method for treating or preventing obesity. In this method, an effective amount of a composition as described above is administered to a mammal, for example, a human. The present invention also provides a method for weight control. In this method, an effective amount of a composition as described above is administered to a mammal, for example, a human. Such administration need not be carried out by a physician or under medical supervision, and may simply include the mammal ingesting the composition, for example, in the form of a food product or food supplement. Preferably, the compositions of the invention are administered (or taken) 2 hours to 3 minutes, more preferably 1 hour to 15 minutes, and even more preferably 35 to 25 minutes before consuming a food. The invention is applicable to mammals, preferably humans.

Other mammals that can benefit from the compositions of the invention, include pets (e.g., dogs, cats, horses, rabbits, hamsters and guinea pigs), and farm animals (e.g., cattle, sheep and pigs). Dogs and cats are particularly preferred. When a food product is produced, the pinolenic acid or derivative thereof can be first mixed with structuring components for use in food products. However, this is not essential. These mixtures can be applied, for example, beneficially in food products as healthy fat compositions. Pine nut oil can be used in the compositions of the invention. However, since pine nut oil may contain up to about 26 wt.% Pinolenic acid, it would be advantageous (in particular for use in food supplements, and to allow dosage forms of a smaller size), if they could obtain concentrates with higher levels of pinolenic acid. Concentrates of pinolenic acid or a derivative thereof which will be used in the present invention can be prepared by any suitable method. A suitable method is described in EP-A-1088552. In a suitable procedure, an enzymatic hydrolysis or glycerolysis is carried out using an enzyme that can discriminate between fatty acids with a delta-5 double bond and other fatty acids. This process comprises: i) reacting a glyceride material containing at least 2% by weight of fatty acid with cis5 double bond with water or glyceroi, in the presence of an enzyme capable of discriminating between fatty acids containing a delta double bond 5 and other fatty acids; ii) separating the reaction mixture into a component rich in partial glycerides and a component rich in fatty acids; iii) optionally converting the partial glycerides of step ii) to free fatty acids in the presence of a suitable enzyme; iv) optionally converting the fatty acid-rich component from step ii) to triglycerides by reaction with glycerol in the presence of a suitable catalyst, such as a suitable enzyme; v) optionally separating the partial glyceride-rich material from step ii) into components that are a) rich in monoglycerides, b) rich in diglycerides, and c) rich in triglycerides, and then optionally converting the partial glycerides a) and b) into triglycerides by reaction with fatty acids in the presence of a suitable enzyme. It is preferred to use a glyceride material with a pinolenic acid content of 5 to 50% by weight, preferably 10 to 35% by weight in step i). Examples of such materials are pinologenic oils, and concentrates thereof. This process produces a concentrate containing at least 28% by weight of pinolenic acid. Enzymes suitable for use in steps i), iii), iv) and v), are lipases. Suitable lipases include Candida rugosa lipase; lipase QL; lipase SL, lipase OF; Rhizopus delemar lipase, Rhizopus oryzae lipase; lipase B from Geotríchum candidum; and Rhizomucor miehei lipase. Preferred enzymes for step i) are lipase from Candida rugosa and lipase B from Geotrichum candidum. Suitable lipases also include Lipozyme IM (a commercial enzyme). The preferred enzyme for use in step iv) is Lipozyme M (from Rhizomucor miehei). The compositions of the invention can be food products. Food products in which pinolenic acid or derivatives thereof can be used, or mixtures containing these compounds include, but are not limited to, margarines; low fat pasta; pasta with very low fat content; bicontinuous pasta; watery continuous pastes; confectionery products, such as chocolates, toppings or fillings; ice creams; ice cream toppings; ice cream inclusions; dressings; mayonnaises; sauces; baking fats; fats; cheese; meat replacement products; healthy bars; muesli bars; drinks; dairy products; low-carbohydrate products; low-calorie products; soups; cereals; and milkshakes The addition of pinolenic acid or a derivative thereof or mixtures containing at least one of the compounds to food products can have a positive effect on texture, hardness, appearance, rheology, oral melting, impact on taste, extensibility, microstructure (crystal size, drop size), aeration properties or ease of processing of these food products. The use of a glyceride of pinolenic acid is particularly advantageous in this respect.

Other examples of compositions are pharmaceutical compositions, such as in the form of tablets, pills, capsules, caplets, multiple particulate matter including granules, beads, pellets and microencapsulated particles; powders, elixirs, syrups, suspensions and solutions. The pharmaceutical compositions will comprise a pharmaceutically acceptable diluent or carrier. The pharmaceutical compositions are preferably adapted for parenteral administration (eg, orally). Orally administrable compositions may be in solid or liquid form, and may take the form of tablets, powders, suspensions and syrups. Optionally, the compositions comprise one or more coloring and / or flavoring agents. Pharmaceutically acceptable carriers suitable for use in such compositions are well known in the pharmacy art. The compositions of the invention may contain from 0.1 to 99% by weight of pinolenic acid. The compositions of the invention are generally prepared in unit dosage form. Preferably, the unit dosage of pinolenic acid is from 1 mg to 1000 mg (more preferably from 100 mg to 750 mg). The excipients used in the preparation of these compositions are the excipients known in the art. Other examples of product forms for the composition, are food supplements (whose term includes nutritional products), such as the form of a soft gel or a hard capsule comprising an encapsulating material selected from the group consisting of gelatin, glycerol, starch, modified starch, and starch derivatives such as glucose, sucrose, lactose and fructose. The encapsulating material may optionally contain crosslinking or polymerizing agents, stabilizers, antioxidants, light absorbing agents to protect light sensitive fillers, preservatives, and the like. Preferably, the unit dosage of pinolenic acid in the food supplements is from 1 mg to 1000 mg (more preferably from 100 mg to 750 mg). The amount of pine nut oil that is used in a unit dosage form is preferably from 100 mg to 2000 mg, for example, from 250 mg to 1500 mg (eg, 750 mg), for example, to be taken four times a day. The compositions of the invention may contain other additives that are well known in the food art, and pharmaceuticals including, but not limited to, flavoring ingredients, coloring agents, sweeteners and emulsifiers. The following non-limiting examples illustrate the invention, and in no way limit its scope. In the examples and throughout this specification, all percentages, parts and ratios are by weight, unless otherwise indicated.

EXAMPLES EXAMPLE 1 An in vitro test was carried out to measure the effect of several free fatty acids on the release of CCK from intestinal cells. The study showed the effect of free oil fatty acids from pine nuts and thus pinologenic acid as a satiety ingredient (see data below).

Cell culture The STC1-1 cell line was cultured at 37 ° C in an atmosphere of 5% CO 2 and 95% air, in the medium of RPMI 1640 supplemented with 5% FCS, 2 mM glutamine, 100 U / ml of penicillin and streptomycin at 50 μM. The cells were routinely passed after reaching 70 to 80% confluence, washing the cell layer with PBS, and incubating with a trypsin-EDTA solution. The seeding density of 2 × 10 6 cells per 75 cm 2 was used for routine subculturing.

Experimental protocol 24 hours before the experiments, STC1 cells were seeded in 6 cavity culture dishes at 40 to 50% confluence. On the day of the experiment, the cells were washed in 1 ml of pH buffer of Krebs-Ringer's bicarbonate (KRBB), pH 7.4, containing BSA at 0.2% (in w / v). The STC1 cells were preincubated for 15 minutes in 2 ml of KRBB, before being incubated in an amount of 2 ml of KRBB, with or without the agents tested, for 1 hour. At the end of the incubation, the supernatant was collected, centrifuged at 1000 rpm for 5 minutes, and frozen immediately at -20 ° C for RIA. The DNA content was measured by fluorometry after extraction of the contents of the culture cells.

RIA analysis The immunoreactivity of CCK was measured using antisera .39A (1 / 300,000) that cross-reacts 100% with CCK-33 and CCK-8, 12% with sulfated gastrin-17, 5% with non-sulfated gastrin-17, and less than 0.1% with CCK-8 and gastrin-1. 34 not sulfated. CCK-8 is used as standard. The Bolton-Hunter CCK-9 (Thr, Nle) was labeled with iodine 125 by the chloramine T method, and purified by inverted phase HPLC. The test pH regulator was 0.05 M sodium phosphate, pH 7.5. Aliquots of 1 μl to 200 μl of supernatant were tested in duplicate (day 0). The mark is added on day 1, and precipitation with carbon is carried out 48 hours later. Figure 1 is a bar graph showing the levels of CCK produced using free fatty acids (FFAs) from pine nuts, and other FFAs. Figure 1 shows that pine nut oil produced a markedly higher level of CCK than the other materials tested. Each sample had a CCK level determined at three different concentrations, which were 100 μM, 500 μM and 1 mM based on a molecular weight for the FFA of 300 g / mol. Marinol is a fish oil concentrate from Loders Crokiaan BV (Wormerveer, The Netherlands) that contains EPA and DHA. Clarinol is a product of Loders Crockiaan BV (Wormerveer, The Netherlands) containing 80% conjugated linoleic acid. The other FFA samples are from a range of different natural products.

EXAMPLE 2 The following is an example is a filled gelatin capsule in accordance with the invention. A composition comprising pinolenic acid is encapsulated in a gelatin capsule according to methods well known in the art. The resulting encapsulated product contains 500 mg of pinolenic acid, and one capsule can be taken up to four times daily by an adult human.

EXAMPLE 3 The following figures of the drawings are referred to in example 3, and are summarized as follows: Figure 2 shows levels of CCK in blood (pmol / L) after ingestion of 3 g of pinolenic acid FFA (graphic above interrupted line) or placebo (lower graph, solid line); Figure 3 shows levels of GLP1 in blood (pmol / L) after ingestion of 3 g of Finolenic acid FFA (upper graph, interrupted line) or placebo (lower graph, solid line); Figure 4 shows the desire to eat for 4 hours after the ingestion of 3 g of Finolenic acid FFA (lower graph, interrupted line) or placebo (upper graph, solid line); Figure 5 shows the prospective food ration for 4 hours after the ingestion of 3 g of pinolenic acid FFA (lower graph, interrupted line) or placebo (upper graph, solid line); Figure 6 shows the levels of CCK in blood (pmoles / L) after ingestion of 3 g of TG of pinolenic acid of example 3 (ii) (upper graph, interrupted line) or placebo (lower graph, solid line); Figure 7 shows the desire to eat for 4 hours after the ingestion of 3 g of TG of pinolenic acid (lower graph, interrupted line) or placebo (upper graph, solid line); and Figure 8 shows the prospective food ration for 4 hours after the ingestion of 3 g of TG of pinolenic acid (lower graph, interrupted line) or placebo (upper graph, solid line). In the figures, an asterisk indicates a significant difference. Two compositions comprising pinolenic acid were tested: example 3 (i), a composition comprising pinolenic acid in the form of free fatty acid (FFA); and Example 3 (ii), a composition comprising pinolenic acid in the form of triglyceride (TG). The fatty acid methyl ester (FAME) analysis of the two compositions is as follows: Four. Five- A double-blind, placebo-controlled, crossover, randomized study was conducted with 18 overweight women (BMI = 25-30) who received 3 g of FFA composition from Example 3 (i), 3 g of TG composition of example 3 (¡) or 3 g of placebo (olive oil) in capsules, in combination with a light breakfast of low fat consisting of two slices of white bread and jam. At 0, 30, 60, 90, 120, 180 and 240 minutes, blood samples were taken for analysis of CCK, GLP-1, glucose, insulin, free fatty acids and triglycerides. Subjective sensations of satiety were evaluated using visual analog scales (VAS) for desire to eat, and prospective food consumption. Each subject received all three treatments in a randomized order within a two-week period with a one-week rest period, according to a Latin square design. The VAS scales consisted of horizontal lines of 150 mm with sentences anchored at each end that expressed the most positive or negative sensation. Subjects traced a vertical line at the point on the horizontal line that corresponded to their feeling of hunger. The distances on the VAS scales were converted to scores between 0 and 100. The concentrations of CCK and GLP-1 in the blood samples were measured using optimized and validated commercial human RIA equipment. The treatments were evaluated statistically with ANOVA, and were considered as significant with a P <; 0.05.

Adverse effects (AEs) were monitored. Ingestion of 3 g of FFA composition of Example 3 (i) induced a maximum value in the release of CCK after 30 minutes, which was significantly greater than the release of CCK in response to placebo (Figure 2). The release of GLP-1 peaked at 60 minutes after ingestion of the FFA composition of Example 3 (i), and was also significantly higher than the level of GLP-1 in response to placebo (Figure 3). Over a period of 4 hours, the total amount of CCK released into the blood stream in response to ingestion of the FFA composition of Example 3 (i) was 60% higher, and for GLP1 levels, it was 25% higher than the placebo, measured by the areas under the curve. The FFA composition of Example 3 (i) also decreased the "desire to eat" and the "prospective ingestion" scores during the 4 hours after ingestion (Figures 4 and 5). These scores were lower at 30 minutes after ingestion of the FFA composition of Example 3 (i), and the differences with placebo were significant. The TG composition of Example 3 (ii) affected appetite sensations and the release of CCK, similarly to the FFA composition of Example 3 (i) (Figures 6 to 8). In summary, the CCK and GLP1 data showed clear and significant treatment effects. The FFA and TG compositions increased satiety levels - inducing hormones, CCK and GLP1 in the blood within 30 to 60 minutes, and the levels continued to be higher for up to 4 hours after ingestion. In addition, the desire to eat and the prospective ingestion scores at 30 minutes were lower after ingestion of the FFA compositions of example 3 (i) and TG of example 3 (ii), than after the placebo. No adverse events or serious adverse events were reported. In addition, the levels of glucose, insulin, free fatty acids and triglycerides in the blood were measured, measured from 0 to 4 hours after the ingestion of the capsules, and showed that the levels of triglycerides and free fatty acids did not vary significantly at any point. of time between the groups. Glucose and insulin levels increased in both groups due to the ingestion of the light breakfast containing sugar that all participants consumed. In all groups, responses to glucose and insulin during the 4 hours were similar, indicating that the FFAs of Example 3 (i) and the TGs of Example 3 (ii) were well tolerated. However, glucose levels did not increase much after ingestion of the FFA composition of example 3 (i) that after ingestion of the TG composition of example 3 (ii) and placebo, and decreased more gradually after of the ingestion of the FFA composition of example 3 (i) and the ingestion of the TG composition of example 3 (ii) than after the placebo (olive oil). As a result, the insulin level increased more slowly, and then decreased rapidly after ingestion of the FFA composition of Example 3 (i) than after the placebo, and after ingestion of the TG composition of Example 3 (ii) ). In this way, the FFA composition of example 3 (i) caused more moderate blood level changes in glucose and insulin than placebo, the TG composition of example 3 being intermediate (ii).

EXAMPLE 4 The release of relative CCK by many different compounds was determined in an in vitro test to measure the effect of various free fatty acids on the release of CCK from intestinal cells. The study showed the effect of oil-free fatty acids of pine nuts, and thus pinolenic acid, as a satiety ingredient (see data below). Other fatty acids present in high and low amounts in the oil of pine nuts (such as oleic acid, linoleic acid, taxoleic acid, sciadonic acid and juniperonic acid), are not capable of inducing high amounts of CCK. A composition with 27% pinolenic acid induces slightly higher amounts of CCK with 16% pinolenic acid.

Cell culture The enteroendocrine STC1-1 cell line was cultured at 37 ° C in an atmosphere of 5% CO 2 and 95% air, in standard culture medium (DMEM). The cells were routinely passed after reaching 70 to 80% confluence, washing the cell layer with PBS, and incubating with a trypsin-EDTA solution. A seeding density of 2 × 10 6 cells per 75 cm 2 was used for routine subculturing.

Experimental Protocol STC1 cells were seeded in 6-well culture plates, and incubated with control culture medium, with or without the agents tested, for 1 hour. All agents were tested at a concentration of 100 μM in the free fatty acid form. Since the fatty acids were diluted in ethanol, the effect of ethanol was tested, and the CCK levels of the reference line were indicated. Capric acid was used as a negative control fatty acid, because it is already known that capric acid does not induce CCK. The effects of bombesin were used as a positive control. At the end of the incubation, the supernatant was collected, centrifuged and frozen immediately at -20 ° C for RIA. The viability of the cells was verified by microscopic analysis, analyzing the DNA content, which was measured by fluorometry after the extraction of the content of culture cells, and in addition the LDH release was measured.

RIA analysis The release of CCK was measured using a RIA of standard CCK, and the effects of fatty acids were measured 6 times. The following table shows that the samples with 16% and 27% of pinolenic acid, respectively, produced an almost 4-fold and 5-fold higher level of CCK than the negative control fatty acid, capric acid. An oil derived from Pinus pinea containing less than 1% pinolenic acid, was less able to induce release of CCK (only about twice as much as capric acid). Other fatty acids present in the oil of pine nuts, such as oleic acid, linoleic acid, taxoleic acid, juniperonic acid and sciadonic acid, were also not able to induce high amounts of CCK (almost twice as much as capric acid). In contrast, other 18: 3 fatty acids, such as punic acid, gamma-linoleic acid, alpha-linoleic acid, and alpha-elefesaric acid, are good inducers of CCK in a similar way to pinolenic acid, but it is noteworthy that these fatty acids are they measure in a pure form, whereas pinolenic acid was tested only at 16% and 27% purity. 95% pure CLA (a 50:50 mixture of c9, t11 and t10, c12 isomers) was not able to induce high amounts of CCK in a CLA-like manner with pinolenic acid. A commercial product that consisted of a mixture of fractionated palm oil and oat oil was also not very active to induce CCK (twice as much as capric acid).

EXAMPLE 5 TG compositions of pinolenic acid and pinolenic acid FFA compositions of examples 3 (i) and 3 (¡) are produced, using "crude pine nut oil" as raw material. Pine nuts are crushed by suppliers in an ejector, applying high pressure at room temperature. The only heat generated during the ejection is caused by the crushing of the seeds in the ejector, and never reaches more than 50 ° C. The extracted oil is then filtered by passing it through canvas. The product obtained is "crude pine nut oil". The crude oil is further processed to obtain the TG and FFA compositions. The processing consists of the following main steps: refining, hydrolysis and distillation. The refining step is a physical refining consisting of a bleaching step using rare bleaching earth that removes residues of contaminants, followed by a deodorization step (steam distillation). The oil of refined pine nuts, is the composition of TG. The refined pine nut oil can be further hydrolyzed using a quality lipase suitable for food, to obtain free fatty acids, glycerol and not fully hydrolyzed oil residues (di- and tri-glycerides). Finally, the hydrolyzed mixture is distilled to obtain the FFA composition.

EXAMPLE 6 Soft gel capsules are produced by rotary matrix processing. The material for the outer cover of the capsules, the gel and the filling, are formulated separately. Once the gel mass and the mass of the filling are ready, the gel is spread in a thin film to form two gelatin strips which are then rolled over two separate matrices which determine the size and shape of the capsules. As the gelatin films are adapted to the matrices, the filling is carefully dosed at a level of 500 mg, 750 mg or 1000 mg of oil per capsule, and injected between the two gelatin strips that are sealed after immediately applying heat and pressure. The capsules fall from the machine, and then dry under a stream of hot air.

Claims (16)

NOVELTY OF THE INVENTION CLAIMS

1. - The use of pinolenic acid or a derivative thereof, in the manufacture of a composition for weight control, reducing the feeling of hunger and / or increasing satiety.

2. The use claimed in claim 1, wherein the composition is in the form of a food supplement, a pharmaceutical composition or a food composition.

3. The use claimed in claim 1 or 2, wherein the composition comprises pinolenic acid as a free fatty acid, a mono-, di- or tri-glyceride, or a mixture thereof.

4. The use claimed in any of claims 1 to 3, wherein the composition is pine nut oil, or is derived from pine nut oil.

5. The use claimed in claim 1, wherein the derivative is selected from alpha-linolenic acid, gamma-linolenic acid, punicic acid, eleastearic acid, and alkyl salts or esters thereof.

6. The use claimed in claim 5, wherein the alkyl esters are mono-, di- or tri-glycerides, or mixtures thereof.

7. The use claimed in any of the preceding claims, wherein the composition further comprises a fatty acid or derivative thereof selected from the group consisting of linoleic acid, oleic acid, conjugated linoleic acid, mixtures of enriched isomers of acid conjugated linoleic, EPA and DHA, or a mixture thereof.

8. The use claimed in claim 7, wherein the fatty acid or derivative thereof is present as a free fatty acid, or a mono-, di- or tri-glyceride, or a mixture thereof.

9. The use claimed in claim 1, wherein the composition further comprises at least one glyceride formed from linoleic acid, oleic acid, trans acids and saturated fatty acids, and the composition is a food composition.

10. The use claimed in claim 9, wherein the glyceride is selected from liquid oils selected from soybean oil, sunflower oil, rapeseed oil and cottonseed oil; cocoa butter and cocoa butter equivalents; palm oil and fractions thereof; enzymatically obtained fats; fish oils and fractions thereof; conjugated linoleic acid and mixtures of enriched isomers thereof; gamma-linoleic acid and enriched mixtures thereof; hardened liquid oils; and mixtures thereof.

11. The use claimed in any of the preceding claims, wherein the composition is a food product selected from the group consisting of: margarines; low fat pasta; pasta with very low fat content; bicontinuous pasta; watery continuous pastes; confectionery products, such as chocolates, toppings or fillings; ice creams; ice cream toppings; ice cream inclusions; dressings; mayonnaises; sauces; baking fats; fats; cheese; meat replacement products; healthy bars; muesli bars; drinks; dairy products; low-carbohydrate products; low-calorie products; soups; cereals; and milkshakes

12. The use claimed in any of claims 1 to 8, wherein the composition is a pharmaceutical composition in a form selected from the group consisting of tablets, pills, capsules, caplets, multiple particulate matter including: granules , beads, pellets and microencapsulated particles; powders, elixirs, syrups, suspensions and solutions.

13.- The use claimed in any of the claims 1 to 8, wherein the composition is a food supplement in the form of a soft gel or a hard capsule comprising an encapsulating material selected from the group consisting of gelatin, starch, modified starch, and starch derivatives.

14. The use claimed in any of the preceding claims, wherein the composition is for weight control, helping to maintain body weight and / or reduce body weight and / or help to thin the body and / or reduce the ingestion of calories and / or allow less room for high-calorie foods.

15. The use claimed in any of claims 1 to 14, wherein the mammal is a human.

16. The use of a composition as defined in any of claims 1 to 14, in the manufacture of a composition for stimulating the production of cholecystokinin (CCK) in a mammal.