Composition comprising L-leucine and fish oil

Abstract

The present invention relates to a composition comprising L-leucine and fish oil, which composition can be used to supplement leucine and [omega] 3 in a subject in need thereof. The invention also relates to the use of said composition in the treatment of muscle tissue pathologies, in particular the use of said composition for the prevention of muscle pathologies characterized by loss of muscle mass and / or muscle strength, such as sarcopenia.

Description

Field of the Invention

[0001] The present invention relates to a composition comprising l-leucine and fish oil, which can be used to supplement leucine and omega 3 in subjects in need thereof.

[0002] The present invention also relates to the use of the composition in treating muscle tissue pathologies, in particular the use of the composition for preventing muscle pathologies characterized by loss of muscle mass and / or muscle strength, such as sarcopenia. Background Art

[0003] L-Leucine is an amino acid that belongs to the group of the so-called essential amino acids, ie those which cannot be synthesized by the organism and therefore have to be taken up from the diet.

[0004] The primary function of L-leucine is as a precursor for protein synthesis and is particularly important at the muscle level as it directly stimulates myofibrillar muscle protein synthesis.

[0005] Leucine has been found to slow down muscle tissue degeneration by increasing muscle protein synthesis and therefore helping to maintain muscle mass and strength.

[0006] Leucine is an aliphatic branched-chain amino acid, nonpolar and poorly soluble in water. Two enantiomers of leucine can be identified: 2(S)-amino-3-methylvaleric acid and 2(R)-amino-3-methylvaleric acid. From an optical point of view, these two molecules are defined as L-leucine and D-leucine. The L-enantiomer is used by organisms to produce proteins.

[0007] Omega-3 also plays a beneficial role in muscle mass.

[0008] ω3 (n-3 PUFA) are polyunsaturated fats that are considered essential. In fact, their precursor (α-linolenic acid, ALA) cannot be synthesized by the body and must therefore be ingested from food.

[0009] The main animal sources of omega 3 are fatty fish, such as anchovies, herring, mackerel, salmon, sardines, sturgeon, trout, cod and tuna, and the oils prepared from them. These oils are particularly rich in the omega 3 EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid), which are easily used by the body once ingested in the diet.

[0010] The composition comprising L-leucine and fish oil can be advantageously used to supplement nutrients necessary for muscles.

[0011] However, even in the presence of leucine, the characteristic fishy smell of fish oil makes its consumption unappealing.

[0012] To overcome this problem encountered with fish oils, formulations have been proposed over time, such as those coated with capsules, which aim to facilitate the intake of omega 3, but unfortunately, these solutions do not solve the unpleasant fishy smell of the oils, which remains in the mouth even after ingestion, causing bad breath; in some cases, the fishy smell can also be smelled on the skin.

[0013] In order to overcome the problem of the adverse organoleptic properties of fish oil, attempts have been made to use plant-based ω3 sources, such as walnuts, flax seeds, chia seeds and their oils, in addition to soybean oil. These sources are particularly rich in ALA, which must first be converted to EPA and DHA in order to be utilized by the body. However, some recent studies (Lane et al., 2014) report that, in reality, ALA from plant sources is actually converted to EPA and DHA very little, or not at all.

[0014] Patent application EP3511010 describes a tasteless fish oil obtained by mixing gum, starch or methylcellulose with water at high temperature (about 80°C-85°C) to obtain a colloid, which is then mixed with fish oil to obtain a tasteless emulsion. However, this preparation procedure is complicated and requires energy consumption to heat the mixture to high temperature. In addition, in the industrial preparation of compositions containing oils, problems are encountered related to the processability of the mixture. Leucine also encounters these processability problems, because this amino acid is poorly soluble in water.

[0015] Thus, compositions comprising leucine and fish oil suffer from processability problems, and even more so when gums are included in order to improve the organoleptic properties of the product.

[0016] It is therefore an object of the present invention to provide a composition comprising L-leucine and fish oil which has pleasant organoleptic properties and is adequately processible so as to overcome the problems and limitations of the prior art. SUMMARY OF THE INVENTION

[0018] This object has been achieved by a composition comprising L-leucine, fish oil, polyoxyethylene sorbitan monooleate (or polysorbate) and at least one gum, the composition being as defined in the appended claims.

[0019] The invention also relates to a food or dietary supplement comprising the composition according to the invention and a suitable food carrier, preferably in the form of a powder or granules to be reconstituted in water for oral administration.

[0020] In another aspect, the invention relates to the use of a composition and a food or a supplement according to the invention for treating muscle tissue pathologies, such as muscle atrophy or sarcopenia. DETAILED DESCRIPTION OF THE INVENTION

[0022] Therefore, the present invention relates to a composition comprising L-leucine, fish oil, polyoxyethylene sorbitan monooleate and at least one gum, wherein the weight ratio of gum to fish oil is higher than 0.3 and lower than 0.8, and wherein the weight ratio of polyoxyethylene sorbitan monooleate to L-leucine is higher than 0.01 and lower than 0.06.

[0023] For the purposes of the present invention, the term "L-leucine" refers indifferently to pure enantiomer L-leucine or to enantiomer L-leucine mixed with preferably lower concentrations than L-leucine, and more preferably at most trace amounts of D-leucine.

[0024] Preferably, the L-leucine according to the present invention is pure L-leucine, ie L-leucine which does not contain the enantiomer D-leucine.

[0025] Fish oils suitable for use in the composition according to the invention include fish oils obtained from anchovies, herring, mackerel, salmon, sardines, sturgeon, trout, cod and tuna as well as other fish oils obtained according to prior art methods.

[0026] The term "gum" generally denotes a natural or synthetic polymeric material characterized by high elasticity. For the purposes of the present invention, the term "gum" means a natural gum comprising or consisting of a polysaccharide.

[0027] Examples of gums that may be used in the present invention are gum arabic, acacia gum, xanthan gum, ghatti gum, tragacanth gum, fenugreek gum, mesquite gum or mixtures thereof.

[0028] Preferably, the at least one gum is selected from gum arabic, xanthan gum, mesquite gum and mixtures thereof.

[0029] In a preferred embodiment, the composition of the present invention comprises gum arabic, or a mixture of gum arabic and xanthan gum.

[0030] Preferably, the weight ratio between gum and fish oil is between 0.4 and 0.7, more preferably between 0.5 and 0.6.

[0031] Preferably, the weight ratio between polyoxyethylene sorbitan monooleate and L-leucine is between 0.02 and 0.05, more preferably between 0.02 and 0.03.

[0032] In fact, it has surprisingly been found that the selection of these specific ingredients in the specified amounts and ratios allows to achieve satisfactory processability at the production stage, as well as equally pleasant organoleptic properties, adequately masking the smell and flavour of fish oil.

[0033] Surprisingly, it was also observed that the use of water-soluble flavors alone did not reduce the fishy smell and taste of the oil, whereas the addition of fat-soluble flavors (fat flavors) to the composition was able to improve its organoleptic properties, further increasing palatability. Therefore, preferably, the composition according to the invention also comprises at least one fat-soluble flavor, optionally in combination with at least one water-soluble flavor.

[0034] Fragrances that may be used in preferred compositions according to the present invention include orange + carrot + lemon (ACE) butter flavor, mint butter flavor, mint flavor, lemon flavor and mixtures thereof.

[0035] Preferably, the composition also comprises at least one sweetener to further improve its palatability.

[0036] The composition may also optionally contain an excipient.

[0037] The term "excipient" means a compound or mixture of compounds suitable for pharmaceutical or food use, respectively. For example, an excipient used in a pharmaceutical or food formulation should generally not cause adverse reactions in a subject, nor should it significantly inhibit the efficacy of the cholecalciferol contained therein.

[0038] Suitable excipients are acidulants, acidity regulators, anti-caking agents, antioxidants, fillers, resistance agents, gelling agents, coating agents, modified starches, chelating agents, thickeners, sweeteners, diluents, deagglomerators, glidants, colorants, binders, lubricants, stabilizers, adsorbents, wetting agents, flavoring agents, film formers, emulsifiers, wetting agents, release retardants and mixtures thereof.

[0039] In some embodiments, the composition according to the invention consists essentially of L-leucine, fish oil, polysorbate and at least one gum. The expression "consists essentially of" means that the additional components in the composition do not interfere with the effects and properties of the specified ingredients.

[0040] In other embodiments, the composition according to the invention consists of L-leucine, fish oil, polysorbate and at least one gum and suitable excipients. The expression "consisting of" means that no further ingredients are present in the composition.

[0041] It is also to be understood that all aspects stated above as favourable and advantageous for the composition and its components are to be considered equally preferred and advantageous for those embodiments defined by the expressions "consisting essentially of" and "consisting of", respectively.

[0042] The compositions of the present invention can be prepared using prior art methods. In practice, for administration via the oral route, the components can be mixed, for example, as is, or mixed with one or more excipients, sealed in a soft gel capsule or in a solid form, such as tablets, minitablets, microtablets, granules, microparticles, pellets, multi-particulates, micronized particles, powders, or in the form of solutions, emulsions, gels, ampoules or drops or sprays.

[0043] Preferably, the composition of the invention is in solid form.

[0044] When the composition is in solid form, the composition may further comprise one or more coating agents.

[0045] The term "coating agent" refers to a film-forming polymer capable of coating the surface of a solid form, thereby forming an outer layer that not only allows the release of the ingredient to be regulated but also further masks the flavor of the fish oil and leucine.

[0046] Suitable film-forming polymers are cellulose acetate phthalate, cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate, polyvinyl acetate phthalate, hydroxyethylcellulose phthalate, cellulose acetate tetrahydrophthalate, methacrylate-methacrylic acid copolymers, methyl methacrylate-methacrylic acid copolymers, sodium alginate, stearic acid, cellulose acetate trimellitate, shellac, hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, ethyl cellulose, cross-linked carboxymethyl cellulose and its salts, polyvinyl alcohol, pH-independent polymethacrylates, polyvinyl acetate, polyvinyl pyrrolidone, fatty acids, medium chain triglycerides, or mixtures thereof.

[0047] Preferably, the coating agent comprises ethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, cellulose, fatty acids, medium chain triglycerides or mixtures thereof.

[0048] When the composition in solid form comprises at least one coating agent, the composition may advantageously be prepared by a process comprising the following steps:

[0049] i) weighing L-leucine, sieving, and then placing the L-leucine in a mixer for about 15 minutes;

[0050] ii) spraying an emulsion comprising gum, fish oil, polyoxyethylene sorbitan monooleate, water and optionally a fat flavor onto the L-leucine and mixing,

[0051] iii) adding an aqueous solution of the coating agent with stirring and allowing the solid form thus coated to dry.

[0052] In all embodiments, the compositions according to the invention were found to taste palatable and pleasant, have excellent organoleptic properties, be stable and easy to prepare and manufacture.

[0053] The invention also relates to a dietary supplement or food comprising the composition according to the invention and a suitable food carrier.

[0054] Preferably, suitable food carriers are sweeteners, stabilizers, thickeners, acidulants, colorants, flavors, fillers and mixtures thereof.

[0055] Preferably, the supplement or food according to the invention comprises a filler, such as calcium phosphate, calcium carbonate, mannitol, maltodextrin or a mixture thereof.

[0056] Preferably, the dietary supplement or food according to the present invention is in solid form, such as tablets, minitablets, microtablets, granules, microparticles, pellets, multiparticulates, micronized particles, powders or mixtures thereof.

[0057] Advantageously, the supplement or food according to the invention may also contain probiotics, vitamins, mineral salts, carbohydrates, proteins, phytonutrients and any other ingredients that generally have a supplementary function.

[0058] In a particularly advantageous embodiment, the supplement or food according to the invention comprises at least one probiotic.

[0059] Preferably, the probiotics suitable for the supplement or food according to the invention belong to the Lactobacillus genus or the Bifidobacterium genus.

[0060] Even more preferably, the probiotic belongs to the genus Lactobacillus.

[0061] In a particularly preferred embodiment, the supplement or food product according to the invention comprises at least one probiotic of the L. paracasei species.

[0062] In a particularly preferred embodiment, the supplement or food comprises L-leucine, fish oil, polyoxyethylene sorbitan monooleate, gum and a probiotic belonging to the genus Lactobacillus, wherein:

[0063] The gum and the fish oil have a weight ratio higher than 0.3 and lower than 0.8, preferably from 0.4 to 0.7, more preferably from 0.4 to 0.5, and

[0064] The weight ratio of polyoxyethylene sorbitan monooleate to L-leucine is higher than 0.01 and lower than 0.06, preferably from 0.02 to 0.05, more preferably from 0.02 to 0.03.

[0065] In a particularly advantageous embodiment of the invention, the supplement or food is in powder form and comprises fat flavors, L-leucine, gum arabic, fish oil, polyoxyethylene sorbitan monooleate, a coating agent (preferably comprising hydroxypropyl cellulose, hydroxypropyl methylcellulose or a mixture thereof), probiotics of the genus Lactobacillus, sucralose, water-soluble flavors, citric acid, calcium phosphate, xanthan gum and mannitol.

[0066] In a preferred embodiment, the composition according to the invention and likewise the dietary supplement or food according to the invention is in unit dosage form.

[0067] Preferably, the unit dose contains 2 g to 10 g of the composition, more preferably 4 g to 6 g, most preferably about 5 g of the composition.

[0068] For example, the composition according to the invention and likewise the dietary supplement or food according to the invention may be in the unit dosage form of a powder or granules.

[0069] Advantageously, said unit dose in powder or granules is contained in a single-dose sachet.

[0070] In another aspect, the present invention relates to the use of a composition and likewise a supplement or food for treating muscle tissue pathologies such as muscle atrophy or sarcopenia. For the purposes of the present invention, the term "treatment" means administering a composition, food or supplement according to the invention to a subject at risk of or already affected by a muscle tissue pathology, with the goal of improving the overall condition of their muscle tissue, as well as with the goal of slowing down, alleviating, reducing and / or preventing any changes in the function of their muscle tissue, in particular preventing and counteracting the loss of muscle mass and muscle strength.

[0071] Preferably, the muscle tissue pathology is muscle atrophy or sarcopenia.Risk factors for the development or worsening of muscle tissue pathology include physiological aging.

[0072] Sarcopenia is a syndrome characterized by a progressive and systemic loss of skeletal muscle mass and strength (muscle wasting), and it is strongly associated with physical disability, poor quality of life, and death.

[0073] In clinical practice, case identification can be initiated when a patient reports symptoms or signs of sarcopenia (i.e., falls, feeling weak, slow walking speed, difficulty getting up from a chair, or weight loss / muscle wasting).

[0074] Sarcopenia can be diagnosed using standardized questionnaires or by measuring handgrip strength using a calibrated handheld dynamometer under well-defined testing conditions, together with interpretative data from an appropriate reference population.

[0075] Preferably, in the treatment of muscular tissue pathologies, the composition and likewise the supplement or food must be administered by the oral, buccal or sublingual route, preferably by the oral route.

[0076] More preferably, the composition and likewise the supplement or food is in solid form and has to be reconstituted before administration and more preferably has to be dissolved in water before administration.

[0077] In a preferred embodiment, the solid form is a powder or granules.

[0078] More preferably, in the treatment of muscle tissue disorders, the composition and likewise the supplement or food must be administered in a daily dose sufficient to provide 1 g to 5 g of L-leucine and up to 1 g of fish oil.

[0079] It is to be understood that all possible combinations of preferred aspects of the compositions and supplements or foods comprising said compositions, and the corresponding uses as set out above are described herein and are therefore also preferred.

[0080] It will also be appreciated that all aspects identified as preferred and advantageous for the compositions according to the invention must be considered to be similarly preferred and advantageous for the preparation and use of said compositions and likewise for dietary supplements and foods.

[0081] The following illustrative, non-limiting examples are provided in support of the present invention. Example

[0082] Example 1. Processability Evaluation

[0083] Various compositions containing gum arabic or mesquite gum and fish oil were prepared and it was determined which compositions provided acceptable processability. The test results are summarized in Table 1a below:

[0084] Table 1a

[0085]

[0086] It is clear that tests have shown that the weight ratio between gum and fish oil must be above 0.3 and below 0.8 to be acceptable. In fact, when this ratio is above a value of 0.7, the viscosity of the solution increases, and at 0.8, the solution almost turns into a gel and can no longer be sprayed. On the contrary, when this value is below 0.3, the solution is no longer stable.

[0087] Then the test was carried out at different weight ratios of polyoxyethylene sorbitan monooleate and leucine. The tests carried out are shown in Table 1b below:

[0088] Table 1b

[0089]

[0090] From the tests conducted, it can be seen that the ratio between polyoxyethylene sorbitan monooleate and leucine must be above 0.01 and below 0.06 to have acceptable processability. If this value is below 0.01, a "cap effect" is produced, with all the leucine being on the surface instead of being dispersed in the aqueous solution. Conversely, a ratio above 0.06 will produce a foaming solution.

[0091] Example 2. Evaluation of sensory properties

[0092] Once acceptable ranges for the ratios of gum / fish oil and polyoxyethylene sorbitan monooleate / L-leucine had been determined to achieve good processability during the industrial production process, experiments were conducted with different types and combinations of flavors to further mask the smell and taste of the fish oil.

[0093] Typical formulations with different types of flavor combinations having a gum / fish oil ratio of about 0.53 and a polyoxyethylene sorbitan monooleate / L-leucine ratio of about 0.023 were tested, as shown in Table 2 below:

[0094] Table 2

[0095]

[0096] Tests have shown that fat flavoring alone is able to mask both the smell and taste of fish oil. Furthermore, by adding water-soluble flavorings in addition to fat-soluble flavorings, even more pleasant results are obtained from a palatability perspective.

[0097] Example 3. Preparation of the composition according to the present invention

[0098] The compositions were prepared as shown in Table 3 below:

[0099] Table 3

[0100] Components Weight (g) Fat spices 0.12 L-Leucine 2.50 Gum Arabic 0.31 fish oil 0.59 Polyoxyethylene sorbitan monooleate 0.06 Sucralose 0.02 Water soluble spices 0.20 Citric Acid 0.10 Calcium phosphate 0.40 Xanthan gum 0.10 Mannitol 0.44

[0101] The composition is prepared by weighing an appropriate amount of L-leucine, and then sieving, mixing in a mixer for 15 minutes, and storing the L-leucine in a suitable container. A uniform emulsion comprising gum arabic, fish oil, polyoxyethylene sorbitan monooleate, fat-soluble spices and water is then sprayed onto the leucine. This step is performed while the mixture obtained is in constant, uniform motion.

[0102] Finally, once properly weighed, the remaining ingredients are added and mixed until a composition in the form of a powder or granules is obtained.

[0103] Example 4. Preparation of the composition according to the present invention

[0104] The compositions were prepared as shown in Table 4 below:

[0105] Table 4

[0106] Components Weight (g) Fat spices 0.12 L-Leucine 2.50 Gum Arabic 0.31 fish oil 0.59 Polyoxyethylene sorbitan monooleate 0.06 Coating agents (including hydroxypropyl cellulose and hydroxypropyl methylcellulose) 0.23 Sucralose 0.02 Water soluble spices 0.20 Citric Acid 0.10 Calcium phosphate 0.40 Xanthan gum 0.10 Mannitol 0.44

[0107] The composition is prepared by weighing an appropriate amount of L-leucine, and then sieving, mixing in a mixer for 15 minutes, and storing the L-leucine in a suitable container. A uniform emulsion comprising gum arabic, fish oil, polyoxyethylene sorbitan monooleate, fat-soluble spices / flavorings and water is then sprayed onto the leucine. This step is performed while the mixture obtained is in constant, uniform motion.

[0108] Subsequently, an aqueous solution of coating agents (comprising hydroxypropylcellulose and hydroxypropylmethylcellulose) was added to the mixture also under constant uniform motion.

[0109] Finally, once properly weighed, the remaining ingredients are added and mixed until a composition in the form of a powder or granules is obtained.

[0110] Example 5. Preparation of food according to the invention

[0111] A food product comprising the composition of Example 4 and probiotics was prepared as shown in Table 5 below:

[0112] Table 5

[0113]

[0114] The food is prepared by weighing an appropriate amount of L-leucine, and then sieving, mixing in a mixer for 15 minutes, and storing the L-leucine in a suitable container. Then a uniform emulsion comprising gum arabic, fish oil, polyoxyethylene sorbitan monooleate, fat spices and water is sprayed onto the leucine. This step is carried out while the mixture obtained is constant and uniformly moving.

[0115] Subsequently, an aqueous solution of coating agents (comprising hydroxypropylcellulose and hydroxypropylmethylcellulose) was added to the mixture, still under constant uniform motion.

[0116] Finally, once properly weighed and mixed, add the remaining ingredients, taking care to add the probiotics last to minimize mechanical stress.

[0117] The compositions according to the invention of Examples 3 and 4 and the food product of Example 5 were found to be tasty and pleasant tasting, to have excellent organoleptic properties, and to be stable and easy to prepare and manufacture.

Claims

1. A composition comprising L-leucine, fish oil, polyoxyethylene sorbitan monooleate and at least one gum, wherein the weight ratio of gum to fish oil is higher than 0.3 and lower than 0.8, and wherein the weight ratio of polyoxyethylene sorbitan monooleate to L-leucine is higher than 0.01 and lower than 0.

06.

2. The composition according to claim 1, wherein the at least one gum is selected from gum arabic, xanthan gum, mesquite gum and mixtures thereof, preferably the at least one gum is gum arabic, or a mixture of gum arabic and xanthan gum.

3. A composition according to claim 1 or 2, wherein the weight ratio of gum to fish oil is from 0.4 to 0.7, preferably from 0.5 to 0.

6.

4. The composition according to any one of claims 1 to 3, wherein the weight ratio of polyoxyethylene sorbitan monooleate to L-leucine is 0.02 to 0.05, preferably 0.02 to 0.

03.

5. The composition according to any one of claims 1 to 4, further comprising fat-soluble flavors and optionally water-soluble flavors.

6. The composition according to any one of claims 1 to 5, further comprising a coating agent, wherein the coating agent preferably comprises ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, cellulose, fatty acid, medium chain triglyceride or a mixture thereof.

7. A dietary supplement or food comprising the composition according to any one of claims 1 to 6 and a suitable food carrier.

8. The dietary supplement or food product of claim 7, further comprising at least one probiotic.

9. The composition according to any one of claims 1 to 6 or the dietary supplement or food according to any one of claims 7 to 8, in a solid form selected from the group consisting of tablets, minitablets, microtablets, granules, microparticles, pellets, multiparticulates, micronized particles and powders.

10. The composition, dietary supplement or food according to claim 9, in the form of a unit dose, the unit dose comprising 2g-10g of the composition according to any one of claims 1-6, preferably 4g-6g of the composition according to any one of claims 1-6.

11. The composition according to any one of claims 1 to 6 or the dietary supplement or food according to any one of claims 7 to 10 for use in treating muscle tissue pathology.

12. The composition for use, dietary supplement or food product according to claim 11, wherein the muscle tissue disorder is sarcopenia.

13. Composition for use, dietary supplement or food according to claim 11 or 12, administered via the oral, buccal or sublingual route, preferably when dissolved in water.

14. A composition for use, a dietary supplement or a food product according to any one of claims 11 to 13, administered in a daily dose so as to provide 1 g to 5 g of L-leucine and up to 1 g of fish oil.

Images