Composition for preventing or treating dyslipidemia or fatty liver comprising ursodioxycholic acid and protamine

Abstract

The present invention relates to a composition for the prevention or treatment of dyslipidemia or fatty liver comprising ursodeoxycholic acid and protamine. When a mixture of ursodeoxycholic acid and protamine according to the present invention is administered, it has an excellent effect of significantly reducing blood triglycerides and alleviating the symptoms of non-alcoholic fatty liver disease, and can be used as a pharmaceutical and / or food composition for the prevention, improvement, and treatment of dyslipidemia or fatty liver-related diseases.

Description

Technology Field

[0001] The present invention relates to a composition for the prevention or treatment of dyslipidemia or fatty liver comprising ursodeoxycholic acid and protamine. Background Technology

[0003] Dyslipidemia refers to a condition in which total cholesterol, LDL cholesterol, and triglycerides in the blood are elevated, or HDL cholesterol is decreased. Hyperlipidemia, a type of dyslipidemia, is a state in which blood triglyceride and cholesterol levels are elevated; it primarily occurs due to excessive fat accumulation or impaired metabolic activity within the body. If blood triglyceride and cholesterol levels remain high, the risk of atherosclerotic or cerebrovascular diseases increases significantly, which can lead to other complications and potentially fatal consequences. Consequently, various hyperlipidemia treatments are being developed to address this condition, primarily focusing on therapies that regulate blood cholesterol and triglyceride levels. Hyperlipidemia treatments can be broadly classified into cholesterol biosynthesis inhibitors and cholesterol absorption inhibitors, with types including statins and fibrates. Statins, the most widely used class of drugs, can reduce blood LDL or cholesterol levels by inhibiting HMG-CoA reductase, but their efficacy is limited. Cholesterol absorption inhibitors can reduce blood cholesterol levels by blocking cholesterol absorption, thereby preventing cholesterol accumulation in the body, and increasing its consumption. However, cholesterol absorption inhibitors can cause side effects by also inhibiting the absorption of other beneficial components of the human body, such as vitamins, and can lead to side effects such as diarrhea due to malabsorption. Therefore, there is a need to develop drugs suitable for treating hyperlipidemia.

[0004] Ursodeoxycholic acid (3α,7β-dihydroxy-5β-cholanoic acid, hereinafter referred to as 'UDCA') has a molecular weight of 392.58 and a molecular formula of C 24 As H4OO4, it possesses pharmacological activities that cleanse the micro-bile ducts in vivo, thereby expelling waste products and toxic bile acids from the micro-bile ducts; stabilize hepatocyte membranes and protect hepatocytes; increase hepatic blood flow; inhibit cholesterol absorption and biosynthesis; dissolve gallstones and inhibit their formation; and normalize immune activity. Consequently, it is used for major clinical indications including cholelithiasis and biliary tract diseases, chronic liver disease and improvement of liver function, and indigestion and fatty liver after small bowel resection (Republic of Korea Registered Patent No. 10-0115555). Although UDCA is one of the components of animal bile, it is almost absent in human bile; instead, chenodeoxycholic acid (CDCA), an optical isomer of UDCA, exists in bile mainly conjugated with glycine and taurine.

[0005] Meanwhile, protamine is a strongly positively charged basic protein that binds to DNA and is found in the sperm nuclei of vertebrates. Protamine is known to protect genetic information by compressing and shielding DNA. Although the amino acid sequence of protamine varies somewhat depending on the species, arginine residues account for approximately two-thirds of the total amino acids. Primarily extracted from fish testes, protamine is widely used in food preservatives and other applications due to its antibacterial activity against common bacteria. Furthermore, due to its strong positive charge, protamine forms a complex with insulin when mixed with it, regulating insulin release and potentially increasing the duration of insulin action. In addition, protamine is used as a modulator in cardiac and vascular surgeries to neutralize the anticoagulant effects of heparin. In addition, protamine sulfate can be used as an antidote for medically used heparin overdose, and it has been reported that protamine effectively reduces bitter / intense astringent tastes when added to beverages containing catechins and inhibits lipase, an enzyme involved in fat absorption. (Japanese Patent Application Publication No. 05-339168)

[0006] As such, ursodeoxycholic acid is mainly used alone or in combination with other vitamins for the treatment and prevention of liver disease, and protamine is also used as a food preservative, insulin release regulator, and heparin neutralizer. However, there have been no reports to date that ursodeoxycholic acid and protamine can be effective in treating dyslipidemia or fatty liver-related diseases by reducing triglycerides, and in particular, no research results have been reported anywhere showing that blood triglycerides are significantly reduced when the two are used in combination. Prior art literature

[0008] Japanese Patent Application Publication No. 05-339168, Republic of Korea Registered Patent No. 10-0115555 The problem to be solved

[0009] The present invention aims to provide a pharmaceutical or food composition for the treatment of dyslipidemia or fatty liver. means of solving the problem

[0010] To achieve the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of dyslipidemia comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine or a pharmaceutically acceptable salt thereof.

[0011] In the present invention, the pharmaceutical composition for preventing or treating dyslipidemia may be characterized by having an effect of reducing blood triglycerides.

[0012] In the present invention, the pharmaceutically acceptable salt of the protamine may be characterized as being a protamine sulfate.

[0013] In the present invention, the pharmaceutical composition for the prevention or treatment of dyslipidemia may be characterized as being a composition for oral administration.

[0014] In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof may be mixed to form a nanocomposite.

[0015] The present invention also provides a pharmaceutical composition for the prevention or treatment of fatty liver comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine or a pharmaceutically acceptable salt thereof.

[0016] In the present invention, the pharmaceutically acceptable salt of the protamine may be characterized as being a protamine sulfate.

[0017] In the present invention, the pharmaceutical composition for the prevention or treatment of fatty liver may be characterized as being a composition for oral administration.

[0018] In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof may be mixed to form a nanocomposite.

[0019] The present invention also provides a food composition for preventing or improving dyslipidemia comprising ursodeoxycholic acid or a food-grade acceptable salt thereof and protamine or a food-grade acceptable salt thereof.

[0020] In the present invention, the food composition for preventing or improving dyslipidemia may be characterized by having an effect of reducing blood triglycerides.

[0021] In the present invention, the food-grade acceptable salt of the protamine may be characterized as being protamine sulfate.

[0022] In the present invention, the ursodeoxycholic acid or a food-grade acceptable salt thereof and the protamine or a food-grade acceptable salt thereof may be mixed to form a nanocomposite.

[0023] The present invention also provides a food composition for preventing or improving fatty liver, comprising ursodeoxycholic acid or a food-grade acceptable salt thereof and protamine or a food-grade acceptable salt thereof.

[0024] In the present invention, the food-grade acceptable salt of the protamine may be characterized as being protamine sulfate.

[0025] In the present invention, the ursodeoxycholic acid or a food-grade acceptable salt thereof and the protamine or a food-grade acceptable salt thereof may be mixed to form a nanocomposite. Effects of the invention

[0026] When a mixture of ursodeoxycholic acid and protamine according to the present invention is administered, it has an excellent effect of significantly reducing blood triglycerides and alleviating symptoms of non-alcoholic fatty liver disease, and can be used as a pharmaceutical and / or food composition for the prevention, improvement, and treatment of dyslipidemia or fatty liver-related diseases. Brief explanation of the drawing

[0028] Figure 1 is a TEM image of the structure of a nanoparticle complex formed by mixing ursodeoxycholic acid and protamine. Figure 2 shows the results of confirming the cytotoxicity of a mixture of ursodeoxycholic acid and protamine using colon cells. Figure 3 shows the results of observing changes in body weight after administering distilled water, protamine, ursodeoxycholic acid, and a mixture of ursodeoxycholic acid and protamine to dyslipidemia mice, respectively. Figure 4 shows the results of administering distilled water, protamine, ursodeoxycholic acid, and a mixture of ursodeoxycholic acid and protamine to dyslipidemia mice, respectively, and checking the serum low-density lipoprotein (LDL) levels. Figure 5 shows the results of administering distilled water, protamine, ursodeoxycholic acid, and a mixture of ursodeoxycholic acid and protamine to dyslipidemia mice, respectively, and checking the blood glucose concentration. Figure 6 shows the results of administering distilled water, protamine, ursodeoxycholic acid, and a mixture of ursodeoxycholic acid and protamine to dyslipidemia mice, respectively, and checking the blood triglyceride levels. Specific details for implementing the invention

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled expert in the art to which this invention pertains. In general, the nomenclature used herein and the experimental methods described below are well known and commonly used in the art.

[0031] In this invention, it was confirmed that when ursodeoxycholic acid and protamine are used in combination, the effect of reducing triglycerides in the blood is excellent, and thus it can exert effects in preventing, improving, and / or treating dyslipidemia.

[0032] Accordingly, in one aspect, the present invention relates to a pharmaceutical composition for the prevention or treatment of dyslipidemia comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine or a pharmaceutically acceptable salt thereof.

[0033] In addition, the present invention relates, in another aspect, to a pharmaceutical composition for the prevention or treatment of fatty liver comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine or a pharmaceutically acceptable salt thereof.

[0034] In the present invention, the pharmaceutical composition may be characterized by having an effect of reducing blood triglycerides, but the effect is not limited thereto.

[0035] In the present invention, the pharmaceutically acceptable salt of the protamine may be characterized as being a protamine sulfate or a chloride salt (-Cl), but is not limited thereto.

[0036] In the present invention, the pharmaceutical composition may be characterized as being a composition for oral administration, but the route of administration is not limited thereto.

[0037] In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof may be mixed to form a nanocomposite.

[0038] In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof may be administered simultaneously or sequentially for the prevention or treatment of dyslipidemia and / or fatty liver.

[0039] In the present invention, the dyslipidemia may be characterized as hyperlipidemia, hypercholesterolemia, or hypertriglyceridemia, but is not limited thereto.

[0040] In the present invention, the fatty liver may be characterized as non-alcoholic fatty liver disease (NAFLD), simple steatosis, steatohepatitis, or hepatocirrhosis, but is not limited thereto.

[0041] In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof may be contained in an amount of 0.1 to 75 weight%, preferably 1 to 50 weight%, based on the total weight of the composition, but is not limited thereto.

[0042] In the present invention, the protamine or a pharmaceutically acceptable salt thereof may be contained in an amount of 0.1 to 75 weight%, preferably 1 to 50 weight%, based on the total weight of the composition, but is not limited thereto.

[0043] In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof may be administered in a weight ratio of 1:0.01 to 10, preferably 1:0.1 to 5, more preferably 1:0.5 to 2, and most preferably 1:1, but are not limited thereto.

[0044] In the present invention, the terms “pharmaceutical composition” or “pharmaceutical formulation” refer to a mixture comprising pharmaceutically acceptable excipients, such as diluents or carriers, that make the composition of the present invention particularly suitable for therapeutic use. According to some embodiments, a pharmaceutical composition comprising the composition of the present invention may be provided by a method of administering a therapeutically effective amount to a subject as needed. In some embodiments, the composition of the present invention may be administered to a human.

[0045] As used in the present invention, "effective amount" or "therapeutically effective amount" refers to an amount of a compound or composition (e.g., a compound or composition of the present invention) sufficient to achieve a beneficial or desired result. The effective amount may be administered in one or more doses, applications, or dosages and is not intended to be limited to a specific formulation or route of administration.

[0046] The pharmaceutical composition provided in the present invention relates, in principle, to a pharmaceutical composition intended for administration to humans; however, a person skilled in the art will understand that such composition is generally suitable for administration to all kinds of animals. That is, the pharmaceutical composition according to the present invention may also be administered to other mammals requiring veterinary treatment, such as livestock (e.g., dogs, cats, etc.), farm animals (e.g., cattle, sheep, pigs, horses, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). A skilled veterinary pharmacologist who has a good understanding of variations of the pharmaceutical composition for administration to various animals can design and / or perform such variations simply through routine experimentation if necessary.

[0047] The pharmaceutical compositions described in the present invention may be prepared by any method known in the pharmacological field or described below. Generally, such methods for tablets include the step of associating an active ingredient with an excipient and / or one or more other auxiliary ingredients, and then, if necessary or desired, forming and / or packaging the product into desired single- or multiple-dose units.

[0048] The pharmaceutical composition of the present invention may be manufactured, packaged, and / or sold unpackaged as a single unit dose and / or multiple single unit doses. As used herein, a “unit dose” is an individual amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient administered to a subject and / or a convenient fraction of such dose, e.g., 1 / 2 or 1 / 3 of the dose.

[0049] The relative amounts of the active ingredient, pharmaceutically acceptable excipients, and / or any additional ingredients in the pharmaceutical composition of the present invention will vary depending on the identity, size, and / or disorder of the patient being treated and the route of administration of the composition. For example, the composition may contain 0.001% to 100% (w / w) of the active ingredient.

[0050] As used in the present invention, pharmaceutically acceptable excipients include any solvent, dispersion medium, diluent, or other liquid vehicle, dispersion or suspension aid, surfactant, isotonic agent, thickener or emulsifier, preservative, solid binder, lubricant, etc. suitable for a specific dosage form. The literature of Remington [The Science and Practice of Pharmacy, 21st Edition, AR Gennaro, (Lippincott, Williams & Wilkins, Baltimore, MD, 2006)] discloses various excipients used in the preparation of pharmaceutical compositions and known techniques for their preparation. Except where any ordinary carrier medium cannot coexist with the substance or its derivatives by, for example, providing any undesirable biological effect or otherwise interacting in a harmful manner with any other component of the pharmaceutical composition, its use is considered to be within the scope of the present invention. Pharmaceutically acceptable excipients are at least 95%, 96%, 97%, 98%, 99%, or 100% pure.

[0051] The excipients are approved for human and veterinary use. In some embodiments, the excipients are approved by the U.S. Food and Drug Administration. In some embodiments, the excipients are pharmaceutical grade. In some embodiments, the excipients meet the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia (EP).

[0052] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersants and / or granulators, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, and / or oils.

[0053] These excipients may optionally be included in the formulation of the present invention. Excipients such as cocoa butter and suppository wax, coloring agents, coating agents, sweeteners, flavoring agents, and perfumes may be present in the composition at the discretion of the pharmacist.

[0054] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dical calcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugars, and combinations thereof.

[0055] Exemplary granulators and / or dispersants include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginate, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponge, cation-exchange resin, calcium carbonate, silicate, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pre-gelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (bi gum), sodium lauryl sulfate, quaternary ammonium compounds, and combinations thereof.

[0056] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and bee gum [magnesium aluminum silicate]), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, and cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethyl Cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate [Twin 20], polyoxyethylene sorbitan [Twin 60], polyoxyethylene sorbitan monooleate [Twin 80], sorbitan monopalmitate [Span 40], sorbitan monostearate [Span 60], sorbitan tristearate [Span 65], glyceryl monooleate, sorbitan monooleate [Span 80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [Mirz 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., cremophor), polyoxyethylene Ether, (e.g., polyoxyethylene lauryl ether [Breeze 30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laureate, sodium lauryl sulfate, Pluronic F 68,Poloxamer 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, sodium docusate, and / or combinations thereof, including but not limited to these.

[0057] Exemplary binders include, but are not limited to, starch (e.g., corn starch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol); natural and synthetic gums (e.g., acacia, sodium alginate, Irish moss extract, Panwer gum, Shaty gum, Isapol husks mucilage, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (V-gum), and larch arabogalactan); alginates; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silica; polymethacrylates; waxes; water; alcohols; and combinations thereof.

[0058] Exemplary preservatives may include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives. Exemplary antioxidants include, but are not limited to, alpha-tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and trisodium edetate. Exemplary antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimid, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexatidine, imidourea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal. Exemplary antifungal preservatives include, but are not limited to, butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid. Exemplary alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol. Exemplary acid preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.Other preservatives include, but are not limited to, tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, fenonib, methylparaben, Germol 115, Germaben II, neolon, carton, and euxyl. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0059] Exemplary buffers include citrate buffer solution, acetate buffer solution, phosphate buffer solution, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentacarbonate, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixture, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixture, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixture, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and combinations thereof, but not limited to these.

[0060] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0061] Exemplary oils include almond, apricot kernel, avocado, babassu, bergamot, black currant seed, borage, cade, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cottonseed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, pumpkin, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, cucui nut, lavandin, lavender, lemon, Lythea cubeba, macadamia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange lapis lazuli, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sazquana, savory, sea buckthorn, sesame, and shea. Butter, silicone, soybean, sunflower, tea tree, milk thistle, tsubaki, vetiver, walnut, and wheat germ oils are included, but not limited thereto. Exemplary oils include butyl stearate, caprylic acid triglyceride, capric acid triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and combinations thereof, but are not limited thereto.

[0062] Liquid formulations for oral and parenteral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, the liquid formulation may include inert diluents commonly used in the art, such as, for example, water or other solvents; solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide; oils (particularly cottonseed, peanut, corn, sprout, olive, castor, and sesame oils); polyethylene glycol and fatty acid esters of glycerol, tetrahydrofurfuryl alcohol, and sorbitan; and mixtures thereof. In addition to the inert diluent, the oral composition may include ajuvants, such as humectants, emulsifiers and suspending agents, sweeteners, flavoring agents, and perfumes. In a specific embodiment for parenteral administration, the pharmaceutical composition of the present invention is mixed with a solubilizing agent such as cremophor, alcohol, oil, denatured oil, glycol, polysorbate, cyclodextrin, polymer, and combinations thereof.

[0063] Injectable formulations, e.g., sterile injectable aqueous or oily suspensions, may be prepared according to the known art using dispersants or wetting agents and suspending agents. Sterile injectable formulations may be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, e.g., solutions in 1,3-butanediol. Among acceptable vehicles and solvents, water, Ringer's solution, USP, and isotonic sodium chloride solutions may be adopted. Additionally, sterile fixing oils are typically adopted as solvents or suspension media. For this purpose, any non-irritating fixing oil containing synthetic mono- or diglycerides may be adopted. Furthermore, fatty acids, such as oleic acid, are used in the preparation of injectable formulations.

[0064] Injectable formulations may be sterilized, for example, by filtration through a bacteria-retaining filter, or by including a sterile agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable media before use.

[0065] To prolong the effect of a drug, it is often desirable to slow down its absorption from subcutaneous or intramuscular injections. This is achieved by using a liquid suspension of a crystalline or amorphous material with low water solubility. The absorption rate of the drug then depends on the solubility, which can ultimately be influenced by crystal size and crystalline form. Alternatively, delayed absorption of parenterally administered drugs is achieved by dissolving or suspending the drug in an oil vehicle.

[0066] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active ingredient comprises one or more inert pharmaceutically acceptable excipients or carriers, such as sodium citrate or dical phosphate and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, such as, for example, carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) humectants, such as glycerol; d) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) dissolution retardants, such as paraffin; f) absorption promoters, such as quaternary ammonium compounds; g) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; h) absorbents, such as kaolin and bentonite clay; and i) lubricants, such as talc, calcium stearate, magnesium stearate, It is mixed with solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof.

[0067] In the case of capsules, tablets, and pills, the dosage forms may include a buffer. Similar types of solid compositions may also be adopted as fillers in soft and hard-filled gelatin capsules using lactose or lacsose, as well as high molecular weight polyethylene glycol, etc., as such excipients. Solid dosage forms of tablets, coated tablets, capsules, pills, and granules may be manufactured with coatings and shells, such as enteric coatings and other coatings well known in the field of pharmaceutical formulation. These may optionally include an opacifying agent and may be compositions that release only the active ingredient, or preferentially, to a specific part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric materials and waxes. Similar types of solid compositions may also be adopted as fillers in soft and hard-filled gelatin capsules using lactose or lacsose, as well as high molecular weight polyethylene glycol, etc., as such excipients.

[0068] The active ingredient may be in a micro-encapsulated form with one or more of the excipients described above. Solid dosage forms such as tablets, coated tablets, capsules, pills, and granules may be manufactured with coatings and shells such as enteric coatings, controlled-release coatings, and other coatings well known in the field of pharmaceutical formulation. In these solid dosage forms, the active ingredient may be mixed with one or more inert diluents, such as sucrose, lactose, or starch. These dosage forms may include additional substances other than inert diluents as in ordinary practice, such as tablet lubricants and other tablet aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may include buffers. They may optionally include opacifiers and may be compositions that release only the active ingredient, or preferentially, to a specific part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymeric materials and waxes.

[0069] Dosage forms for topical and / or transdermal administration of the pharmaceutical composition of the present invention may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and / or patches. Generally, the active ingredient is mixed with a pharmaceutically acceptable carrier and / or any necessary preservative and / or a buffer that may be required under sterile conditions. Additionally, the present invention considers the use of transdermal patches, which have the additional advantage of providing controlled delivery of the active ingredient to a body. Such dosage forms may be prepared, for example, by melting and / or dispersing the active ingredient in a suitable medium. Alternatively, or additionally, the ratio may be controlled by providing a ratio-controlling membrane and / or by dispersing the active ingredient in a polymer matrix and / or gel.

[0070] Formulations for topical administration include, but are not limited to, liquid and / or semi-liquid formulations such as ointments, lotions, oil-in-water and / or water-in-oil emulsions such as creams, ointments / or pastes, and / or solutions and / or suspensions. Although the concentration of the active ingredient may be as high as the limit of the solubility of the active ingredient in the solvent, the topically administered formulation may contain, for example, about 1% to about 10% (w / w) of the active ingredient. The formulation for topical administration may additionally contain one or more additional ingredients as described herein.

[0071] The pharmaceutical compositions of the present invention described herein are typically prepared in the form of dosage units for easy and uniform administration. However, it will be understood that the total daily dosage of the compositions of the present invention will be determined by the attending physician within the scope of reasonable medical judgment. The specific therapeutically effective dose level for any particular subject will depend on various factors including disease, disorder, or the disorder and severity of the disorder being treated; the activity of the specific active ingredient adopted; the specific composition adopted; the subject's age, weight, overall health, gender, and diet; the time of administration, route of administration, and elimination rate of the specific active ingredient adopted; the duration of treatment; drugs used in combination with or simultaneously with the specific active ingredient adopted; and factors well known in the medical field, etc.

[0072] The pharmaceutical composition of the present invention may be administered by any route. In some embodiments, the pharmaceutical composition is administered by various routes including oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, intradermal, rectal, vaginal, intraperitoneal, topical (by powder, ointment, cream, and / or droplet), mucosa, nose, mouth, enteral, sublingual; tracheal drip infusion, bronchial drip infusion, and / or inhalation; and / or oral spray, nasal spray, and / or aerosol. Routes specifically considered are invasive intravenous injection, local administration via blood and / or lymphatic supply, and / or direct administration to the affected area. In general, the most suitable route of administration will depend on various factors including the characteristics of the agent (e.g., stability in the environment of the gastrointestinal tract) and the impairment of the subject (e.g., whether the subject can tolerate oral administration).

[0073] In certain embodiments, the pharmaceutical composition of the present invention may be administered at a dosage level sufficient to deliver about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.1 mg / kg to about 40 mg / kg, about 0.5 mg / kg to about 30 mg / kg, about 0.01 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 10 mg / kg, or about 1 mg / kg to about 25 mg / kg of the subject's body weight at least once a day to achieve the desired therapeutic effect. The target dosage may be delivered three times a day, twice a day, every day, every two days, every three days, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the target dose may be delivered through multiple administrations (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more administrations).

[0074] It will be understood that the dosage ranges described in the present invention provide guidance for the administration of pharmaceutical compositions provided to adults. For example, the amount administered to children or adolescents may be determined by a specialist or a person skilled in the art and may be less or equal to that administered to adults. The exact amount of peptide according to the present invention required to achieve an effective dose will vary from subject to subject, for example, depending on the subject's species, age, and overall disorder, side effects or severity of disorder, identity of the characteristic compound, mode of administration, etc.

[0075] It will be understood that the pharmaceutical compositions of the present invention may be used in combination therapy. A specific combination of treatments (therapeutics or procedures) to be used in combination therapy will be chosen considering the intended therapeutic effect to be achieved and the suitability of the intended therapeutic agents and / or procedures.

[0076] The pharmaceutical composition of the present invention may be administered alone or in combination with one or more therapeutic agents. In the case of "combination," while the following delivery methods fall within the scope of the present invention, it is not intended to imply that the agents must be administered at the same time and / or formulated to be delivered together. The composition may be administered simultaneously with, before, or after one or more other therapeutic agents or medical procedures. Generally, each agent will be administered at a prescribed dosage and / or time schedule for that agent. Additionally, the present invention encompasses delivering the pharmaceutical composition of the present invention in combination with an agent capable of improving its bioavailability within the body, reducing and / or modifying its metabolism, inhibiting its secretion, and / or modifying its distribution. It will be further understood that the pharmaceutical composition and therapeutic agent of the present invention used in this combination may be administered together as a single composition or separately as different compositions.

[0077] The specific combination used in combination therapy will take into account the intended therapeutic effect to be achieved and / or the suitability of the procedure and / or therapeutic agent comprising the peptide of the present invention. It will be understood that the combination used may achieve the intended effect for the same disorder (e.g., the pharmaceutical composition of the present invention may be administered in combination with another therapeutic agent (e.g., a second therapeutic agent) used to treat the same disorder), and / or these may achieve different effects (e.g., control of any side effects).

[0078] As used herein, “therapeutic agent” refers to any substance used as a medicine to treat, prevent, delay, reduce, or improve a disorder, and refers to a substance used for treatment, including preventive and curative treatments.

[0079] In some embodiments, the pharmaceutical composition of the present invention may be administered in combination with any therapeutic agent or procedure (e.g., surgery, radiation therapy) useful for treating, alleviating, improving, or mitigating one or more symptoms or features, delaying their onset, inhibiting their progression, reducing their severity, and / or reducing their incidence.

[0080] In one embodiment, the present invention may be provided as a kit for treating said disease in a subject suffering from dyslipidemia and / or fatty liver.

[0081] In some embodiments, the kit comprises i) instructions for administering the pharmaceutical composition according to the present invention to a subject suffering from the disease, and ii) the pharmaceutical composition according to the present invention. In some embodiments, the kit may comprise one or more unit dosage forms containing a dosage of the pharmaceutical composition as described in the present invention that is effective in treating the disease in a subject. In some embodiments, the subject is a human patient.

[0082] In some embodiments, the kit further comprises one or more selected from the group comprising a sterile syringe, a sterile needle, a sterile IV bag, an infusion pump, or any combination thereof.

[0083] The term "about" as used in the present invention may be interpreted as meaning approximately, roughly, or to a certain degree. When the term "about" is used in conjunction with a numerical range, it is interpreted as modifying the range by extending the boundaries above and below the specified numerical value. Generally, the term "about" is used in the present invention to modify numerical values ​​above and below the value specified by a 10% variance.

[0084] "Individual," "patient," and "subject" are used interchangeably and include any animal, including mammals, e.g., mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates including humans.

[0085] In the present invention, "treatment" means reversing, alleviating, inhibiting the progression of, or preventing one or more symptoms of a disease or illness to which the term applies, or said disease or illness, unless otherwise stated; and as used herein, the term "treatment" refers to the act of treating when "treating" is defined as above. Accordingly, "treatment" or "therapeutic regimen" of a disease in mammals may include one or more of the following:

[0086] (1) Preventing the development of the disease,

[0087] (2) Preventing the spread of disease,

[0088] (3) Relieves disease,

[0089] (4) Preventing the recurrence of the disease and

[0090] (5) Palliating the symptoms of the disease

[0091] In the present invention, "prevention" means any act of suppressing or delaying the occurrence, spread, and recurrence of dyslipidemia or fatty liver by administering a pharmaceutical composition according to the present invention, unless otherwise stated.

[0093] In another aspect, the present invention relates to a food composition for preventing or improving dyslipidemia comprising ursodeoxycholic acid or a food-grade acceptable salt thereof and protamine or a food-grade acceptable salt thereof.

[0094] In another aspect, the present invention relates to a food composition for preventing or improving fatty liver, comprising ursodeoxycholic acid or a food-grade acceptable salt thereof and protamine or a food-grade acceptable salt thereof.

[0095] In the present invention, the food composition is characterized by exhibiting an effect of reducing blood triglycerides, but the effect of action is not limited thereto.

[0096] In the present invention, the food-grade acceptable salt of the protamine may be characterized as being protamine sulfate, but is not limited thereto.

[0097] In the present invention, the ursodeoxycholic acid or a food-grade acceptable salt thereof and the protamine or a food-grade acceptable salt thereof may be mixed to form a nanocomposite.

[0098] In the present invention, the dyslipidemia may be hyperlipidemia, hypercholesterolemia, or hypertriglyceridemia, but is not limited thereto.

[0099] In the present invention, the fatty liver may be characterized as non-alcoholic fatty liver disease (NAFLD), simple steatosis, steatohepatitis, or hepatocirrhosis, but is not limited thereto.

[0100] In the present invention, the ursodeoxycholic acid or a food-grade acceptable salt thereof may be contained in an amount of 0.1 to 75 weight%, preferably 1 to 50 weight%, based on the total weight of the composition, but is not limited thereto.

[0101] In the present invention, the protamine or a food-grade acceptable salt thereof may be contained in an amount of 0.1 to 75 weight%, preferably 1 to 50 weight%, based on the total weight of the composition, but is not limited thereto.

[0102] In the present invention, the ursodeoxycholic acid or a food-grade acceptable salt thereof and the protamine or a food-grade acceptable salt thereof may be administered in a weight or volume ratio of 1:0.01 to 10, preferably 1:0.1 to 5, more preferably 1:0.5 to 2, and most preferably 1:1, but are not limited thereto.

[0103] The food composition of the present invention may be a food or a food additive.

[0104] There are no special restrictions on the above food as long as it contains the above food composition, and it may also be used as an additive to other foods. Examples of foods to which the above food composition may be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense.

[0105] The food of the present invention may include functional foods, particularly health functional foods, and the functional food of the present invention may additionally contain various auxiliary ingredients as needed in addition to the active ingredients. In the case of the functional food of the present invention, it may include vitamins such as vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, folic acid, vitamin C, vitamin D3, and vitamin E, as well as minerals such as copper, calcium, iron, magnesium, potassium, and zinc, or lactic acid bacteria.

[0106] In addition, among the functional foods of the present invention, health drinks may include various flavoring agents or natural carbohydrates as additional ingredients, just like ordinary beverages. Flavoring agents may include natural sweeteners such as taumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame. Natural carbohydrates may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.

[0108] Examples

[0109] The present invention will be described in more detail below through examples. These examples are intended solely to illustrate the present invention, and it will be obvious to those skilled in the art that the scope of the present invention is not to be interpreted as being limited by these examples.

[0111] Example 1: Preparation of a mixture of ursodeoxycholic acid and protamine

[0112] Ursodeoxycholic acid (Sigma-Aldrich, U5127) was stored at room temperature. For the experiment, 10 mg was weighed and slowly dissolved in 1 mL of DMSO solvent, followed by sonification for approximately 1 minute to ensure maximum dissolution. For protamine (P3369, Sigma-Aldrich, USP testing specifications), 10 mg was weighed and slowly dissolved in 10 mL of distilled water. The two solutions were slowly mixed in a 1:10 volume ratio, and distilled water was used as an additional solvent to achieve a final concentration of 1 mg / mL. For samples administered to animals, purified water was used as the dilution solution.

[0114] Example 2: Analysis of a mixture of ursodeoxycholic acid and protamine

[0115] The particles of the mixture of ursodeoxycholic acid and protamine prepared in Example 1 were analyzed using a Scanning Transmission Electron Microscope (STEM).

[0116] The sample solution was sonicated for 1 minute immediately before measurement, and then 10 μL of the mixture prepared in Example 1 was applied onto a TEM grid (HC400-Cu-100 Holey Carbon 400 Mesh) and dried at 65°C for about 8 hours. The microstructure of the sample was analyzed using an electron beam with an EVO MA 10 (Carl Zeiss) scanning electron microscope.

[0117] As a result, as shown in Figure 1, it was observed that a mixture of ursodeoxycholic acid and protamine formed a nanocomposite with a size of 30 to 300 nm.

[0119] Example 3. Analysis of cytotoxicity of a mixture of ursodeoxycholic acid and protamine

[0120] To evaluate the potential cytotoxicity of the ursodeoxycholic acid and protamine mixture, E. coli cells (CT26, Korean Cell Line Bank) were cultured for one week at 37°C under 5% CO2 conditions in DMEM medium (Sigma-Aldrich) supplemented with 10% FBS. Subsequently, cells were detached using trypsin-EDTA solution and transferred to a 96-well plate at a rate of 1 × 10⁶ cells. 3 After dispensing into cells / wells, a mixture of 10 or 100 µM ursodeoxycholic acid and protamine was administered. After 24 hours of incubation, cytotoxicity was calculated by adding 10 µL of EZ-Cytox (Dogenbio) solution and measuring the absorbance at 450 nm, according to the EZ-cytox measurement protocol provided by the manufacturer.

[0121] As a result, as shown in Figure 2, no significant cytotoxicity was observed when the ursodeoxycholic acid and protamine mixture was treated compared to the control group that was not treated.

[0123] Example 4. Effect of improving blood lipid levels in a mouse model of dyslipidemia

[0124] Four-week-old C57BL / 6J mice (Orient Bio, Korea) were supplied, and the following day, their diet was switched to a high-cholesterol diet (Atherogenic Diet, D12336) to construct an animal model of dyslipidemia. The mouse model described above is an experimental animal model capable of pathologically mimicking dyslipidemia in humans through a high-cholesterol diet. After approximately 10 days of feeding, administration of a mixture of ursodeoxycholic acid and protamine was initiated while maintaining the high-cholesterol diet. The mixture of ursodeoxycholic acid and protamine (1:1) was administered daily at 20 mg / kg / day (UDCA 10 mg + Protamine 10 mg). Only purified water was administered as the control group, while the comparative experimental groups used were a group administered protamine alone at 10 mg / kg / day or a group administered ursodeoxycholic acid alone at 10 mg / kg / day. Administration was performed orally daily using an oral sonde for mice, and the dosage per mouse was 200 µl.

[0126] 4-1. Changes in body weight

[0127] As a result of measuring changes in mouse body weight over 28 days, as shown in Figure 3, the protamine-alone group, the ursodeoxycholic acid-alone group, and the ursodeoxycholic acid and protamine mixture group all showed no significant difference in body weight compared to the control group.

[0129] 4-2. Changes in Blood Levels of Low-Density Lipoprotein, Glucose, and Triglycerides

[0130] Blood was collected from mice on the 28th day of administration. The collected blood was left at room temperature for 1 hour, and then centrifuged at 3000 rpm for 20 minutes to obtain serum. Subsequently, blood tests were conducted by sending samples to a testing laboratory (KP&T) to check for changes in low-density lipoprotein, glucose, and triglyceride levels in the blood.

[0131] As for blood low-density lipoprotein, as shown in Figure 4, the levels of low-density lipoprotein (LDL) specifically increased in the group administered protamine alone compared to the control group, while the group administered ursodeoxycholic acid alone and the group administered a mixture of ursodeoxycholic acid and protamine did not show a significant difference in low-density lipoprotein levels compared to the control group.

[0132] As a result of measuring blood glucose levels, as shown in Figure 5, the group administered with a mixture of ursodeoxycholic acid and protamine showed slightly lower values ​​compared to the group administered with distilled water, the group administered with protamine alone, and the group administered with ursodeoxycholic acid alone.

[0133] Meanwhile, as a result of serum triglyceride analysis, as shown in Figure 6, it was confirmed that the blood triglyceride levels of the group administered a mixture of ursodeoxycholic acid and protamine were significantly reduced compared to the group administered distilled water, the group administered protamine alone, and the group administered ursodeoxycholic acid alone.

[0135] Foregoing, specific parts of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents.

Claims

Claim 1 A pharmaceutical composition for the prevention or treatment of dyslipidemia comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine or a pharmaceutically acceptable salt thereof. Claim 2 A pharmaceutical composition according to claim 1, characterized in that the composition exhibits an effect of reducing blood triglycerides. Claim 3 A pharmaceutical composition according to claim 1, characterized in that the pharmaceutically acceptable salt of the protamine is protamine sulfate. Claim 4 A pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition is a composition for oral administration. Claim 5 A pharmaceutical composition according to claim 1, characterized in that the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof are mixed to form a nanocomposite. Claim 6 A pharmaceutical composition for the prevention or treatment of fatty liver comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine or a pharmaceutically acceptable salt thereof. Claim 7 A pharmaceutical composition according to claim 6, characterized in that the pharmaceutically acceptable salt of the protamine is protamine sulfate. Claim 8 A pharmaceutical composition according to claim 6, characterized in that the above pharmaceutical composition is a composition for oral administration. Claim 9 A pharmaceutical composition according to claim 6, characterized in that the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine or a pharmaceutically acceptable salt thereof are mixed to form a nanocomposite. Claim 10 A food composition for preventing or improving dyslipidemia comprising ursodeoxycholic acid or a food-grade acceptable salt thereof and protamine or a food-grade acceptable salt thereof. Claim 11 A food composition according to claim 10, characterized in that the above composition exhibits an effect of reducing blood triglycerides. Claim 12 A food composition according to claim 10, characterized in that the food-grade acceptable salt of the protamine is protamine sulfate. Claim 13 A food composition according to claim 10, characterized in that the ursodeoxycholic acid or a food-grade acceptable salt thereof and the protamine or a food-grade acceptable salt thereof are mixed to form a nanocomposite. Claim 14 A food composition for preventing or improving fatty liver, comprising ursodeoxycholic acid or a food-grade acceptable salt thereof and protamine or a food-grade acceptable salt thereof. Claim 15 A food composition according to claim 14, characterized in that the food-grade acceptable salt of the protamine is protamine sulfate. Claim 16 A food composition according to claim 14, characterized in that the ursodeoxycholic acid or a food-grade acceptable salt thereof and the protamine or a food-grade acceptable salt thereof are mixed to form a nanocomposite.

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