CA3135250C

CA3135250C - Stable emulsified vitamin and omega fatty acid compositions and process for preparing same

Info

Publication number: CA3135250C
Application number: CA3135250A
Authority: CA
Inventors: Behnam RAHMANLOO; Mohammadreza Tahajomi; Sattar Tahajomi; Amirhossein Tahajomi; Meysam Araghi; Alireza Naeimi Panjaki
Original assignee: Zecto Inc
Current assignee: Zecto Inc
Priority date: 2021-10-21
Filing date: 2021-10-21
Publication date: 2024-02-13
Anticipated expiration: 2041-10-21
Also published as: CA3135250A1

Abstract

The present invention provides a stable emulsified oil-soluble vitamin and omega fatty acid composition, oral dosage forms comprising the emulsified composition, and a process for preparing the emulsified composition.

Description

STABLE EMULSIFIED VITAMIN AND OMEGA FATTY ACID COMPOSITIONS AND
PROCESS FOR PREPARING SAME
FIELD OF THE INVENTION
[0001] The present invention pertains to the field of pharmaceutical compositions and in particular to stable emulsions of oil-soluble vitamins and fatty acids.
BACKGROUND

[0002] Vitamins and fatty acids are organic compounds that humans need in small quantities.
Many of them need to come from food because our body either does not produce them or produces very little.

[0003] Vitamins D, E, and K are fat-soluble vitamins. The body stores fat-soluble vitamins in fatty tissue and the liver.

[0004] Vitamin D is a group of fat-soluble secosteroids. Several forms (vitamers) of vitamin D
exist. The two major forms are vitamin D2 or ergocalciferol, and vitamin D3 or cholecalciferol.
Chemically, the various forms of vitamin D are secosteroids, i.e., steroids in which one of the bonds in the steroid rings is broken. The structural difference between vitamin D2 and vitamin D3 is the side chain of D2 that contains a double bond between carbons 22 and 23, and a methyl group on carbon 24. Vitamin D without a subscript refers to either D2 or D3 or both.

[0005] Vitamin D (also referred to as "calciferol") is a fat-soluble vitamin that is naturally present in a few foods, added to others, and available as a dietary supplement. It is also produced endogenously when ultraviolet (UV) rays from sunlight strike the skin and trigger vitamin D synthesis.

Date recue/date received 2021-10-21

[0006] Vitamin D obtained from sun exposure, foods, and supplements is biologically inert and must undergo two hydroxylations in the body for activation. The first hydroxylation, which occurs in the liver, converts vitamin D to 25-hydroxyvitamin D [25(OH)D], also known as "calcidiol." The second hydroxylation occurs primarily in the kidney and forms the physiologically active 1,25-dihydroxyvitamin D [1,25(OH)2D], also known as "calcitriol".

[0007] Vitamin D is essential for strong bones, because it helps the body use calcium from the diet. Traditionally, vitamin D deficiency has been associated with rickets, a disease in which the bone tissue doesn't properly mineralize, leading to soft bones and skeletal deformities. But increasingly, research is revealing the importance of vitamin D in protecting against a host of health problems.

[0008] Vitamin D has been shown to be useful for the prevention and treatment multiple sclerosis, the prevention and treatment type 1 and type 2 diabetes, the prevention and treatment hypertension, the prevention and treatment glucose intolerance, the decreased risk of some types of cancer, and for to decrease the risk of death from cardiovascular disease.

[0009] Vitamin E is a group of eight fat-soluble compounds that include four tocopherols and four tocotrienols. There are also water-soluble derivatives of Vitamin E.
Vitamin E is a fat-soluble vitamin. It is found in many foods including vegetable oils, cereals, meat, poultry, eggs, fruits, vegetables, and wheat germ oil. It is also available as a supplement.

[0010] Vitamin E can be used for treating vitamin E deficiency, as well as for treating, preventing or managing many other conditions such as: Alzheimer disease, low levels of red blood cells in people with long-term illness (anemia of chronic disease), a blood disorder that reduces levels of protein in the blood called hemoglobin (beta-thalassemia), leakage of intravenous (IV) drug from the vein into the surrounding skin and tissue (extravasation), menstrual cramps (dysmenorrhea), scarring or hardening of blood vessels in the kidney (glomerulosclerosis), an inherited disorder that causes red blood cells to break down in response to stress (G6PD deficiency), a type of non-cancerous skin sore (granuloma annulare), an inherited brain disorder that affects movements, emotions, and thinking (Huntington disease), bleeding within the skull (intracranial hemorrhage), bleeding into or around the fluid-filled areas (ventricles) of the brain (intraventricular hemorrhage), conditions in a man that prevent him from getting a woman pregnant within a year of trying to conceive (male infertility), Date recue/date received 2021-10-21 reduced benefit of nitrate therapy that happens when nitrates are used all day (nitrate tolerance), swelling (inflammation) and buildup of fat in the liver in people who drink little or no alcohol (nonalcoholic steatohepatitis or NASH), Parkinson's disease, recovery from laser eye surgery (photoreactive keratectomy), premenstrual syndrome (PMS), physical performance in elderly adults, scarring of tissue caused by radiation therapy, rheumatoid arthritis (RA), movement disorders often caused by antipsychotic drugs (tardive dyskinesia), and swelling (inflammation) of the eye (uveitis).

[0011] "Vitamin K" is the generic name for a family of fat-soluble compounds with a common chemical structure of 2-methyl-1,4-naphthoquinone, that is naturally present in some vegetables like spinach, asparagus, and broccoli or in eggs and also is available as a dietary supplement.

[0012] "Vitamin K" include several chemical compounds. These are similar in structure in that they share a quinone ring, but differ in the length and degree of saturation of the carbon tail and the number of repeating isoprene units in the side chain. Plant-sourced forms are primarily vitamin K1, whereas animal-sourced forms are primarily vitamin K2.

[0013] Vitamin K plays a key role in helping the blood clot, preventing excessive bleeding.
Unlike many other vitamins, vitamin K is not typically used as a dietary supplement.

[0014] Matrix Gla-protein, a vitamin K-dependent protein present in vascular smooth muscle, bone, and cartilage, is the focus of considerable scientific research because it might help reduce abnormal calcification. Osteocalcin is another vitamin K-dependent protein that is present in bone and may be involved in bone mineralization or turnover

[0015] In the circulation, vitamin K is carried mainly in lipoproteins.
Compared to the other fat-soluble vitamins, very small amounts of vitamin K circulate in the blood.
Vitamin K is rapidly metabolized and excreted. This rapid metabolism accounts for the relatively low levels of vitamin K in blood and tissue stores compared to those of the other fat-soluble vitamins.

[0016] Low levels of vitamin K can raise the risk of uncontrolled bleeding.
While vitamin K
deficiencies are rare in adults, they are very common in newborn infants. A
single injection of vitamin K for newborns is standard. Vitamin K is also used to counteract an overdose of the blood thinner coumadin.

Date recue/date received 2021-10-21

[0017] The omega-fatty acids include, for example, omega-3-, omega-6- and omega-9-fatty acids.

[0018] Fatty acids are long-chain (monounsaturated, polyunsaturated, and saturated) acids found mainly in plants and fish. Fatty acids are organic, meaning that they contain both carbon and hydrogen molecules. Chemically, fat molecules are made up of four parts: a molecule of glycerol on one end of the molecule chain, and three molecules of fatty acids on an opposite end of the molecule chain. Each fatty acid consists of a hydrocarbon chain with a carboxyl group at one end. Fatty acids are typically referred to as omega fatty acids and the number following the term "omega-" represents the position of the first double bond, counting from the terminal methyl group on the molecule.

[0019] Omega 3 is a polyunsaturated fatty acid found primarily in fish, nuts and vegetable oils.
The most common recommendation for adults is to eat at least 900 milligrams of it per day.
Children need around half of this amount. It is also useful for pregnant women. There are three types of omega 3: DHA (docosahexaenoic acid), EPA (eicosapentaenoic acid), and ALA (alpha-linolenic acid). Even though the emphasis is usually on the former two, in fact, you require all three. Both lemon juice and fish oil provide the body with numerous minerals and health benefits that strengthen the heart and kidneys and cardiovascular and other systems.

[0020] Pregnant women, in particular, have an increased need for omega-3 fatty acids.
Studies have shown that fatty acids are needed for fetal growth, brain development, learning, and behavior. Thus, lactating women are encouraged to increase their intake of fatty acids since infants receive their essential fatty acids through breast milk as more research is undertaken, people become more aware of the possible health benefits of omega-3 fatty acids and there is added incentive to supplement a person's diet with omega-3 fatty acids. Some medical experts recommend at least 500 mg per day of combined EPA and DHA.

[0021] In addition to omega-3 fatty acids, other fatty acids such as omega-6 and omega-9 fatty acids are also known to provide health benefits. Omega-6 fatty acids are a family of unsaturated fatty acids having in common a final carbon-carbon double bond in the n-6 position;
i.e., the sixth bond from the methyl end of the fatty acid. There are nine members in this family of fatty acids, with the two most important being linoleic acid (LA), which may be found in Date recue/date received 2021-10-21 soybean oil, corn oil, safflower oil, peanut oil, cottonseed oil, and rice bran oil; and Arachidonic acid (AA), which may be found in certain meats, eggs, and dairy products. Like omega-3 fatty acids, omega-6 fatty acids are polyunsaturated fats that cannot be produced in the human body.
Thus, omega-3 and omega-6 fatty acids are classed as essential fatty acids (EFAs) to the human diet that can only be obtained by consuming foods such as meat, poultry and eggs, as well as nut and plant-based oils.

[0022] Omega-9 fatty acids are a family of unsaturated fatty acids having in common a final carbon-carbon double bond in the n-9 position; i.e., the ninth bond from the methyl end of the fatty acid. There are five members in this family of fatty acids, with the two most important being oleic acid (OA), which is commonly found in olive oil, canola oil, and other monounsaturated fats; and erucic acid (EA), which can be extracted from rapeseed, wallflower seed, and mustard seed.

[0023] The most widely available source of fatty acids, namely EPA and DHA, is cold-water oily fish, such as mackerel, herring, tuna, salmon, anchovies, and sardines.
Most people in the Western world do not get enough omega fatty acids in their diet, especially those with low fish diets. Therefore, there is a need to supplement the human diet with the omega fatty acids.

[0024] The omega fatty acids in the form of fish oil are safe for most people when used appropriately. However, fish oil can have a "fishy" aftertaste and can also cause belching, nosebleeds, nausea, and loose stools. A major concern with fish oil is that because fish do not produce the omega fatty acids themselves but accumulate it in their bodies from the ingestion of microalgae and seaweed, the fish oil may contain traces of heavy metals, in particular, mercury and other contaminants. Thus, a need exists for a safe, easily digestible, and palatable dietary supplement that will directly supply a sufficient dosage of the omega fatty acids to the human body, without the drawbacks of fish oil mentioned above.

[0025] As a result, the rate of oxidation of EPA/DHA fatty acids added to the aqueous suspension via the supplementation oil can be reduced at least two-fold or even much more compared to the rate of oxidation of an equal quantity of the same EPA/DHA
fatty acid-containing enriching oil homogenized into the same aqueous suspension without having been first combined and diluted with the oxidative stabilization oil.
Date recue/date received 2021-10-21

[0026] Omegas can be dissolved in normal non-carbonated beverages (drinks) with varying degrees of good stability and shelf life. Historically, omegas when put into a carbonated drink have significant time limited taste and short shelf-life issues. By time limited taste, it is meant that over time the taste changes and becomes unsatisfactory for the taster. By short shelf life, it is mean that in certain cases the carbonated drink product may not survive 48 hours after production.

[0027] US2008/274175 describes emulsions or liquids in which are mixed oils comprising omega-3 fatty acids. These compositions may optionally contain vitamin B3 and iron but without using oil-soluble Vitamins.

[0028] WO 2011/060084 describes stable, water soluble formulations comprising omega-type fatty acids in combination with stabilizing agents and one or more additives selected from metal chelators, water-soluble reducing agents, water-insoluble reducing agents, bisulfite and metabisulfite salts, or mixtures thereof.

[0029] EP3188715B1 describes oil-in-water emulsion comprising omega-3 fatty acids, chelated iron, vitamin B and an emulsifier, however without density control agent.

[0030] Therefore there is a need for emulsions of omega fatty acids and oil-soluble vitamins that are shelf-stable for long periods of time under ambient storage conditions.

[0031] This background information is provided to reveal information believed by the applicant to be of possible relevance to the present invention. No admission is necessarily intended, nor should be construed, that any of the preceding information constitutes prior art against the present invention.
SUMMARY OF THE INVENTION

[0032] An object of the present invention is to provide a stable emulsified vitamin and omega fatty acid compositions and process for preparing same. In accordance with an aspect of the present invention, there is provided an emulsified oil-soluble vitamin and omega fatty acid composition comprising: one or more oil-soluble vitamins; one or more omega fatty acids; at Date recue/date received 2021-10-21 least one emulsifying agent; a density control agent; an antioxidant and/or a preservative agent;
and water, wherein the composition is a stable water-in-oil emulsion or oil-in-water emulsion.

[0033] In accordance with another aspect of the present invention, there is provided an oral dosage form comprising the emulsified composition of the present invention.

[0034] In accordance with another aspect of the present invention, there is provided a process for preparing a stable emulsified oil-soluble vitamin and omega fatty acid composition comprising the following steps: providing a mixture comprising one or more oil-soluble vitamins, one or more omega fatty acids, and an antioxidant and/or preservative agent and blending the combination to provide a first blended mixture; adding at least one emulsifying agent to the first blended mixture and blending to provide a second blended mixture; adding water to the second blended mixture and blending in a high shear mixer to provide a third blended mixture; adding a density control agent to the third blended mixture and blending to provide a fourth blended mixture; and passing the fourth blended mixture through a high-pressure homogenizer to provide the emulsified oil-soluble vitamin and omega fatty acid composition.
DETAILED DESCRIPTION OF THE INVENTION

[0035] The present invention provides a stable, oil-in-water (or water-in-oil) emulsion formulation comprising one or more oil-soluble vitamins; one or more omega fatty acids; at least one emulsifying agent; a density control agent; an antioxidant and/or a preservative agent; and water.

[0036] The term "emulsion" as used herein refers to a lipophilic molecule of the present application emulsified (solubilized) in an aqueous medium using a solubilizing agent of the present application. In one example, the emulsion includes micelles formed between the lipophilic molecule(s) and the solubilizing agent. When those micelles are sufficiently small, the emulsion is essentially clear. Typically, the emulsion will appear clear (e.g., transparent) to the normal human eye, when those micelles have a median particle size of about 100 nm. In one embodiment, the micelles in the emulsions of the present invention have median particle sizes between about 30 nm to about 170 nm, preferably between about 30 nm to about 130 nm, more preferably between about 40 nm to about 100 nm.

Date recue/date received 2021-10-21

[0037] Emulsions are not thermodynamically stable, so achieving stable emulsified compositions is of great commercial interest, particularly oil-in-water emulsions comprising both fat-soluble vitamins and fatty acids that can be stored for long periods of time under ambient conditions. A stable emulsion is one in which separation between the aqueous phase and the lipophilic component does essentially not occur (e.g., the emulsion stays clear).

[0038] The term "micelle" is used herein according to its art-recognized meaning and includes all forms of micelles, including, for example, spherical micelles, cylindrical micelles, worm-like micelles and sheet-like micelles, and vesicles, formed in water, or mostly water.

[0039] The terms "fat-soluble" and "oil-soluble" are used herein interchangeably and refer to molecules that are generally hydrophobic and soluble in nonpolar solvents and insoluble in polar solvents.

[0040] As used herein, the term "weight percent" or "wt%" refers to the weight of a component relative to the weight of the total composition.

[0041] As used herein, the term "about" refers to a +/-10% variation from the nominal value. It is to be understood that such a variation is always included in a given value provided herein, whether or not it is specifically referred to.

[0042] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0043] In accordance with the present invention, the emulsified composition comprises one or more omega fatty acids.

[0044] The term "omega fatty acid" is used herein to define a family of polyunsaturated fats having health benefits, and is intended to include omega-3, -6 and -9 fatty acids. These fats are differentiated by the position of the unsaturation. Useful omega fatty acids include any of those known in the art. In any of the embodiments described herein, the omega fatty acids (e.g., omega-3, omega-6, and/or omega-9 fatty acids) can be used in the form of fatty acids or salts thereof (e.g., free carboxylic acids or sodium salts thereof), esters (e.g., a methyl ester, an ethyl Date recue/date received 2021-10-21 ester, etc.), and/or glycerides (e.g., a triglyceride, a diglyceride, or a monoglyceride). In some embodiments, the omega fatty acids are used in the form of triglycerides. In some embodiments, the omega fatty acids are used in the form of ethyl esters.

[0045] Accordingly, in one embodiment, the one or more omega fatty acids used in the compositions of the present invention are selected from omega-3, -6 and -9 fatty acids, and pharmaceutically acceptable salts and esters thereof, in any combination.

[0046] Omega-3, -6 and -9 fatty acids are obtained from natural sources like rapeseed, almonds, cashews, walnut, soybean, pecans, macadamia nuts, pistachios, peanuts, perilla seeds, sesame seeds, flax seeds, chia seeds, camelina seeds, pine nuts, hemp, sunflower seeds, pumpkin seeds and olive oils or other oils containing eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), alpha-linolenic acid (ALA), gamma-linolenic acid (GLA), dihomo-gamma-linolenic acid (DGLA), conjugated linoleic acid (CLA) and oleic acid.

[0047] In one embodiment, the concentration of omega fatty acids in the emulsion is at least about 1 mg/mL and can be as high as about 500 mg/mL.

[0048] In one embodiment, the composition comprises from about 5 to about 15 wt% of the one or more omega fatty acids.

[0049] In accordance with the present invention, the emulsified composition comprises one or more oil-soluble vitamins. In one embodiment, the oil-soluble vitamins are selected from vitamins D, E and K. Vitamins D, E and K may be obtained from natural or synthetic sources. In a preferred embodiment, vitamins D, E and K are used in their synthetic form.

[0050] As used in the present disclosure, the term "vitamin K" is used to describe several chemical compounds that are similar in structure in that they share common structural feature of a 2-methyl-1,4-naphthoquinone, but differ in the length and degree of saturation of the carbon tail and the number of repeating isoprene units in the side chain. Plant-sourced forms are primarily vitamin K1, whereas animal-sourced forms are primarily vitamin K2.
In accordance with the present disclosure, either form may be employed in the emulsified compositions of the Date recue/date received 2021-10-21 present invention. Vitamin K without a subscript should be interpreted to refer to either K1 or K2 or both.

[0051] Vitamin E is a group of eight fat-soluble compounds that include a-tocopherol, 8-tocopherol, y-tocopherol, 5-tocopherol, a-tocotrienol, 8-tocotrienol, y-tocotrienol, and 5-tocotrienol.

[0052] As used in the present disclosure, unless otherwise specified, the term "vitamin E" may be understood to refer to any of the eight fat-soluble compounds or any combination thereof.

[0053] The various forms of vitamin E may be used as food additive in oily food to deter rancidity caused by peroxidation. Accordingly, in accordance with the present disclosure, vitamin E may be used in the compositions of the present invention as an antioxidant agent.

[0054] Vitamin D is a group of fat-soluble secosteroids, i.e., steroids in which one of the bonds in the steroid rings is broken. The two major forms are vitamin D2 or ergocalciferol, and vitamin D3 or cholecalciferol. The structural difference between vitamin D2 and vitamin D3 is the side chain of D2 that contains a double bond between carbons 22 and 23, and a methyl group on carbon 24. As used in the present disclosure, unless otherwise specified, the term "vitamin D"
without a subscript refers to either D2 or D3 or both.

[0055] In one embodiment, the concentration of vitamins in the emulsion is at least about 0.1 mg/mL and can be as high as about 50 mg/mL.

[0056] In one embodiment, the composition comprises from about 0.03 to about 0.08 wt% of the one or more oil-soluble vitamins.

[0057] In accordance with the present invention, the emulsified composition comprises at least one emulsifying agent/surfactant.

[0058] The terms "surfactant" and "emulsifier" are used herein interchangeably and refer to molecules that have a hydrophobic portion and a hydrophilic portion. A
surfactant is a substance that stabilizes an emulsion by increasing its kinetic stability. A surfactant may be non-ionic, ionic or amphoteric.

[0059] Emulsifying agents/surfactants suitable for this purpose can be chosen from Polysorbate 20 (polyoxyethylene (20) sorbitan monolaurate), Polysorbate 40 (polyoxyethylene Date recue/date received 2021-10-21 (20) sorbitan monopalmitate), Polysorbate 60 (polyoxyethylene (20) sorbitan monostearate), Polysorbate 80 (polyoxyethylene (20) sorbitan monooleate), sorbitan monolaurate (Span 20), sorbitan monostearate (Span 60), sorbitan tristearate (Span 65), lecithin, and polyethylene glycol 1000.

[0060] These emulsifying agents/surfactants have HLB value between 2.1 (for sorbitan tristearate) to 16.7 (for polysorbate 20). In one embodiment, at least one surfactant is used in the emulsified compositions of the present invention. It has been observed that good stability is achieved when a combination of surfactants is used. Accordingly, in a preferred embodiment, a mixture of surfactants is used in the emulsified compositions of the present invention.

[0061] In one embodiment, the concentration of emulsifying agents/surfactants in the emulsion is at least about 1 mg/mL and can be as high as about 20 mg/mL.

[0062] In one embodiment, the composition comprises from about 1 to about 2 wt% of the emulsifying agent.

[0063] In accordance with the present invention, the emulsified composition further comprises antioxidant and/or preservative agents.

[0064] The phrase "antioxidant agent" as used herein means an agent that can prevent or reduce an oxidation, degradation and/or other decomposition that would otherwise occur to components or ingredients of the compositions of the invention, such as the omega fatty acids or the oil-soluble vitamins.

[0065] Examples of suitable antioxidant and/or preservative agents include, but are not limited to, ascorbic acid, citric acid, ascorbic acid palmitate, vitamin E (a-tocopherol, aT) and water-soluble precursors of Vitamin E (a-tocopherol, aT), such as its esters with acetate (aTA), succinate (aTS), or phosphate (aTP), and synthetic antioxidants, such as tertiary butylhydroquinone (TBHQ), butylated hydroxyamisole (BHA), and butylated hydroxytoluene (BHT) and combinations thereof. A wide variety of antioxidant agents are commercially available from sources known by those of skill in the art.

Date recue/date received 2021-10-21

[0066] In one embodiment, the concentration of antioxidant in the emulsified composition is at least about 0.05 mg/mL and can be as high as about 1 mg/mL.

[0067] In one embodiment, the composition comprises from about 0.01 to about 2.0 wt% of the antioxidant.

[0068] In accordance with the present invention, in a preferred embodiment, the emulsified composition comprises a density control agent. As used herein, the term "density control agent"
refers to a substance used to control the density of emulsion in order to ensure stability and minimize the tendency for the different components to separate due to a density differential between micelle and water. In a preferred embodiment, glycerol is employed as a density control agent in the compositions of the present invention. When glycerol is added to the formulation, stability increases and also the size of the resulting micelles decreases.

[0069] In one embodiment, concentration of glycerol in the emulsion is at least about 10 mg/mL
and can be as high as about 1000 mg/mL.

[0070] In one embodiment, the composition comprises from about 1 to about 2 wt% of the density control agent.

[0071] In view of the significant nutritional, medical and/or other health benefits provided by omega fatty acids and oil-soluble vitamins such as vitamins D, E and K, it would be beneficial to provide oral dosage forms for oral administration to mammals that disintegrate upon ingestion, thereby allowing the oils and oil-soluble vitamins to become bioavailable, permitting the oils and oil-soluble vitamins to be absorbed by the body without being substantially or fully degraded in the acidic environment of the stomach, and which have a long shelf life under ambient (room temperature) storage conditions.

[0072] The phrase "oral dosage forms" as used herein means edible dosage forms that are suitable for an oral administration to a mammal. These may include hard or soft gel or other capsules, microcapsules and caplets, and the like, containing the emulsified composition. Such oral dosage forms may include in their outer structure gelatin or some other suitable or Date recue/date received 2021-10-21 conventional material. Such materials are known by those of skill in the art, and are commercially available from sources known by those of skill in the art.

[0073] The oral dosage forms of the present invention may also be provided in the form of a beverage or a liquid supplement, or may be incorporated into any suitable foodstuff.

[0074] In accordance with the present invention, the emulsified compositions are prepared using a stepwise process that ensures the complete mixing and emulsification of the mixture under conditions that do not damage or degrade the individual components to provide a final emulsified composition having the desired long-term stability.

[0075] In a first step, a mixture comprising one or more oil-soluble vitamins, one or more omega fatty acids, and an antioxidant and/or preservative agent is blended in a mixer to provide a first mixture, then the emulsifying agent(s) is added to the first mixture and blended to provide a second mixture. Water is then added to the second mixture, and the resulting mixture is blended in a high shear mixer to provide a third mixture. A density control agent is then added, followed by a final blending step to provide an emulsified mixture, which is then passed through a high-pressure homogenizer to provide the final emulsified oil-soluble vitamin and omega fatty acid composition.

[0076] Care is taken at each step of the process to ensure that the temperature of the mixture does not exceed 40 C, thereby minimizing degradation of the components of the composition, assuring the high quality of the components of the final composition as well as its long-term stability.

[0077] The invention will now be described with reference to specific examples. It will be understood that the following examples are intended to describe embodiments of the invention and are not intended to limit the invention in any way.
EXAMPLES
EXAMPLE 1:

Date recue/date received 2021-10-21

[0078] A mixture of 625 g of combination of fatty acids (flaxseed oil, olive oil), 1g of a mixture of fat-soluble vitamins (2 mg vitamin K, 2 mg vitamin D, and 0.98 g vitamin E) and 1 g of ascorbic acid palmitate was mixed at 1000 rpm for 5 minutes with an IKA Eurostar 400 control overhead mixer, then 100 g of a surfactant blend (50g Tween 20, 40g Lecithin, 10g Span 65) was added and the resulting mixture was mixed at 1000 rpm for 10 minutes. Then 5000 gram of water was added, and the resulting mixture was mixed in a stainless steel container at 2000 rpm for 10 minutes with a high shear SIIverson L5M-A mixer. The temperature was checked throughout the process to ensure that it never exceeded 40 C. 100 gram of glycerol was then added and the resulting mixture was mixed for another 10 minutes.

[0079] The resulting emulsion was passed one time through a high-pressure homogenizer (Homolab 2.20 laboratory homogenizer from FBF ITALIA S.r.I.) under 20000 psi pressure. The homogenizer included a cooling system in order to control temperature, to ensure that it did not exceed 40 C.

[0080] Then stability of the resulting emulsion was tested by spinning a sample of the emulsion in a centrifuge at 3000 rpm for 30 minutes, and by incubating a sample of the emulsion in an oven at a temperature of 50 C for 60 days.

[0081] DLS tests showed that the size of micelles in the resulting emulsion were below 100 nm.
No changed in the size of the micelles was observed after centrifugation. The emulsion was also observed to be stable after 60 days in the oven and no separation between the two phases of the emulsion occurred.
EXAMPLE 2:

[0082] The emulsions were prepared using the process as described in Example 1, except the amount of the fatty acids was varied. The amount of fatty acid and the resulting micelle size in the emulsions are summarized in Table 1.
Table 1:

Date recue/date received 2021-10-21 Amount Emulsion size Type (g) (nm) rapeseed, oleic acid 600 85 chia, camelina, olive oil 725 100 sunflower, olive oil 650 90 EPA, DHA, olive oil 300 65 alpha-linolenic acid (ALA), olive oil 300 60

[0083] The stability of the resulting emulsions was tested in a centrifuge at 3000 rpm for 30 minutes and also in an oven with temperature of 50 C for 60 days. The resulting micelle size as determined using DLS is shown in Table 1. No change in micelle size was observed after centrifugation. The emulsions were again shown to be stable after 60 days in the oven and no separation between the two phases was observed.

[0084] These results appear to indicate that varying the composition of the fatty acid component does not impact the size of micelles but varying the amount of the fatty acid component in the composition is important. It was also observed that when using pure fatty acids (i.e., not provided as a mixture of fatty acids within an extracted oil), less oil was used and the resulting stability of the final emulsions and the size of droplets is improved.
EXAMPLE 3:

[0085] The emulsions were prepared using the process as described in Example 1, except the amount of the surfactants was varied. The amounts of each surfactant and the resulting micelle size in the emulsions are summarized in Table 5.
Table 2:
Date recue/date received 2021-10-21 Sorbitan Sorbitan Sorbitan PEG Emulsion Polysorbate Polysorbate Polysorbate Polysorbate LECITHIN
monolaurate monostearate tristearate 1000 size 20 (g) 40 (g) 60 (g) 80 (g) (9) (9) (9) (9) (9) (nm)

[0086] The stability of the resulting emulsions was tested in a centrifuge at 3000 rpm for 30 minutes and also in oven with temperature of 50 C for 60 days. The resulting micelle size as determined using DLS is shown in Table 2. No change in micelle size was observed after centrifugation. The emulsions were also shown to be stable after 60 days in the oven and no separation between the two phases was observed.

[0087] These results appear to indicate that varying the amount of surfactants is important in size of micelles, and that with an increase of the overall relative amount of surfactants, the size of the micelles is smaller. Polyethylene glycol 1000 was observed to have a large effect in reducing the size of the micelles.
EXAMPLE 4:

[0088] The emulsions were prepared using the process as described in Example 1, except the amount of the glycerol was varied. The amounts of glycerol and the resulting micelle size in the emulsions are summarized in Table 3.
Table 3:
Amount Emulsion size (g) (nm) Date recue/date received 2021-10-21

[0089] The stability of the resulting emulsions was tested in a centrifuge at 3000 rpm for 30 minutes and also in an oven with temperature of 50 C for 60 days.

[0090] The resulting micelle size as determined using DLS is shown in Table 3.
No change in micelle size was observed after centrifugation. The emulsions were also shown to be stable after 60 days in the oven and no separation between the two phases was observed.

[0091] These results appear to indicate that varying the amount of glycerol is important in the size of micelles, and that with an increase of the overall relative amount of glycerol, the size of the micelles is smaller.

[0092] It is obvious that the foregoing embodiments of the invention are examples and can be varied in many ways. Such present or future variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

Date recue/date received 2021-10-21

Claims

THE EMBODIMENTS OF THE INVENTION FOR WHICH AN EXCLUSIVE PROPERTY OR
PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. An emulsified oil-soluble vitamin and omega fatty acid composition comprising:
one or more oil-soluble vitamins;
one or more omega fatty acids;
at least one emulsifying agent;
a density control agent;
an antioxidant and/or a preservative agent; and water wherein the composition is a stable water-in-oil emulsion or oil-in-water emulsion.

2. The composition of claim 1, comprising from about 0.03 to about 0.08 wt%
of the one or more oil-soluble vitamins.

3. The composition of claim 1 or 2, wherein the one or more oil-soluble vitamins are selected from a vitamin D compound, a vitamin E compound, a vitamin K compound, and any combination thereof.

4. The composition of claim 3, wherein the vitamin D compound is vitamin D2, vitamin D3 or a combination thereof.

5. The composition of claim 3, wherein the vitamin E compound is selected from a-tocopherol, p-tocopherol, y-tocopherol, 6-tocopherol, a-tocotrienol, 8-tocotrienol, y-tocotrienol, 6-tocotrienol, and any combination thereof.

6. The composition of claim 3, wherein the vitamin K compound is vitamin K1 or vitamin K2.

7. The composition of any one of claims 1 to 6, comprising from about 5 to about 15 wt% of the one or more omega fatty acids.

8. The composition of any one of claims 1 to 7, wherein the one or more omega fatty acids are selected from omega-3 fatty acid, omega-6 fatty acid, omega-9 fatty acid, and pharmaceutically acceptable salts and esters thereof, or a combination thereof.

Date Recue/Date Received 2023-09-05

9. The composition of claim 8, wherein the one or more omega fatty acids are provided in the form of an acid, a salt, a monoglyceride, a diglyceride, a triglyceride, or a combination thereof.

10. The composition of any one of claims 1 to 9, wherein the omega fatty acids are obtained from rapeseed, almonds, cashews, walnut, soybean, pecans, macadamia nuts, pistachios, peanuts, perilla seeds, sesame seeds, flax seeds, chia seeds, camelina seeds, pine nuts, hemp, sunflower seeds, pumpkin seeds or olive oils.

11. The composition of any one of claims 1 to 10, wherein the omega fatty acids are selected from eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), alpha-linolenic acid (ALA), gamma-linolenic acid (GLA), dihomo-gamma-linolenic acid (DGLA), conjugated linoleic acid (CLA) and oleic acid, and combinations thereof.

12. The composition of any one of claims 1 to 11, comprising from about 1 to about 2 wt% of the emulsifying agent.

13. The composition of any one of claims 1 to 12, wherein the emulsifying agent is one or more agents selected from Polysorbate 20, Polysorbate 40, Polysorbate 60, Polysorbate 80, sorbitan monolaurate, sorbitan monostearate, sorbitan tristrearate, lecithin, and polyethylene glycol 1000.

14. The composition of any one of claims 1 to 13, comprising from about 1 to about 2 wt% of the density control agent.

15. The composition of any one of claims 1 to 14, wherein the density control agent is glycerol.

16. The composition of any one of claims 1 to 15, comprising from about 0.01 to about 2.0 wt% of the antioxidant and/or preservative agent.

17. The composition of any one of claims 1 to 16, wherein the antioxidant and/or preservative agent are selected from ascorbic acid, ascorbic acid palmitate, a-tocopherol (aT), a-tocopherol acetate (aTA), a-tocopherol succinate (aTS), a-tocopherol phosphate (aTP), butylated hydroxytoluene (BHT), and combinations thereof.

Date Recue/Date Received 2023-09-05

18. The composition of any one of claims 1 to 17, comprising from about 80 to about 90 wt%
of the water.

19. The composition of any one of claims 1 to 18, wherein the emulsion comprises micelles having a median particle size between about 30 nm to about 170 nm.

20. The composition of any one of claims 1 to 18, wherein the emulsion comprises micelles having a median particle size between about 30 nm to about 130 nm.

21. The composition of any one of claims 1 to 18, wherein the emulsion comprises micelles having a median particle size between about 40 nm to about 100 nm.

22. An oral dosage form comprising the emulsified composition as defined in any one of claims 1 to 21.

23. The oral dosage form of claim 22, wherein the oral dosage form is a hard capsule, a soft gel capsule, a microcapsule or a caplet containing the emulsified composition.

24. The oral dosage form of claim 22, wherein the oral dosage form is in the form of a beverage.

25. The oral dosage form of claim 22, wherein the oral dosage form is in the form of a liquid supplement.

26. A process for preparing a stable emulsified oil-soluble vitamin and omega fatty acid composition comprising the following steps:
providing a mixture comprising one or more oil-soluble vitamins, one or more omega fatty acids, and an antioxidant and/or preservative agent and blending the combination to provide a first blended mixture;
adding at least one emulsifying agent to the first blended mixture and blending to provide a second blended mixture;
adding water to the second blended mixture and blending in a high shear mixer to provide a third blended mixture;
Date Recue/Date Received 2023-09-05 adding a density control agent to the third blended mixture and blending to provide a fourth blended mixture; and passing the fourth blended mixture through a high-pressure homogenizer to provide the emulsified oil-soluble vitamin and omega fatty acid composition.

27.
The process of claim 26, wherein the processing temperature in each step does not exceed 40 C.

Date Recue/Date Received 2023-09-05