KR20230131694A - Composition for preventing or treating dyslipidemia or fatty liver comprising ursodioxycholic acid and protamine - Google Patents

Abstract

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

Description

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

The present invention relates to a composition for preventing or treating dyslipidemia or fatty liver comprising ursodeoxycholic acid and protamine.

Dyslipidemia refers to a state in which total cholesterol, LDL cholesterol, and triglycerides in the blood are increased or HDL cholesterol is decreased. Hyperlipidemia, a type of dyslipidemia, is a condition in which the levels of triglycerides and cholesterol in the blood are elevated and usually occurs when excessive fat accumulation or metabolic activity in the body is not carried out properly. If the levels of neutral fat and cholesterol in the blood remain high, the risk of arteriosclerosis or cerebrovascular disease greatly increases, which can cause other complications, which can lead to fatal consequences for the human body. Therefore, various treatments for hyperlipidemia are being developed to treat this condition, mainly treatments that control blood cholesterol and triglyceride levels. Hyperlipidemia treatments can be broadly divided into cholesterol biosynthesis inhibitors and cholesterol absorption inhibitors, and types include statins and fibrates. The most widely used statin class of drugs can reduce blood LDL or cholesterol levels by inhibiting HMG-CoA reductase, but their efficacy is limited. Cholesterol absorption inhibitors block the absorption of cholesterol, preventing accumulation of cholesterol in the body and increasing consumption, thereby reducing the concentration of cholesterol in the blood. However, in the case of cholesterol absorption inhibitors, they can cause side effects by inhibiting the absorption of other useful ingredients in the human body, such as vitamins, and can cause side effects such as diarrhea due to malabsorption, so the development of drugs suitable for treating hyperlipidemia is difficult. It is necessary.

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 H ₄ 0O ₄ , which acts on microbiliary tract in vivo. It has a cleaning effect, discharging waste products and toxic bile acids in the microbiliary tract, stabilizing the liver cell membrane and protecting liver cells, increasing liver blood flow, inhibiting the absorption and biosynthesis of cholesterol, dissolving gallstones and inhibiting their formation. It has pharmacological activity that normalizes action and immune activity, and is used for major clinical indications in cholelithiasis, biliary tract diseases, chronic liver disease, improvement of liver function, indigestion after small bowel resection, and fatty liver (Korea Registered Patent No. 10-0115555) . UDCA is one of the bile components of animals, but it is almost absent in human bile, and CDCA (chenodeoxycholic acid), an optical isomer of UDCA, is mainly present in bile conjugated with glycine and taurine.

Meanwhile, protamine is a basic protein with a strong positive charge that exists in the sperm nucleus of vertebrates and binds to DNA. Protamine is known to play a role in protecting genetic information by compressing and protecting DNA. The amino acid sequence of protamine is somewhat variable depending on the species, but arginine residues make up about two-thirds of the total amino acids. Protamine, which is mainly extracted from the testes of fish, is widely used in food preservatives, etc. because it exhibits antibacterial activity against common bacteria. In addition, protamine has a strong positive charge, so when mixed with insulin, it forms a complex with insulin and can increase the duration of insulin action by controlling the release of insulin. In addition, protamine is used as a regulator in heart surgery, vascular surgery, etc. to neutralize the anticoagulant effect of heparin. Protamine sulfate can also be used as an antidote for medically used heparin overdose. Protamine can be added to beverages containing catechins to effectively reduce bitterness/intense astringency, and lipase, an enzyme involved in fat absorption. It has been reported to inhibit (Japanese Patent Application Publication No. 05-339168)

Likewise, 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 for purposes such as a food preservative, insulin release regulator, and heparin neutralizer. There has been no report that deoxycholic acid and protamine can be effective in treating dyslipidemia or fatty liver-related diseases by reducing triglycerides. In particular, there are no research results showing that blood triglycerides are significantly reduced when the two are used together. It has not been reported.

Japanese Patent Application Publication No. 05-339168 Republic of Korea Patent No. 10-0115555

The purpose of the present invention is to provide a pharmaceutical or food composition for the treatment of dyslipidemia or fatty liver.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating dyslipidemia comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine and a pharmaceutically acceptable salt thereof. provides.

In the present invention, the pharmaceutical composition for preventing or treating dyslipidemia may be characterized as having a blood neutral fat reducing effect.

In the present invention, the pharmaceutically acceptable salt of protamine may be protamine sulfate.

In the present invention, the pharmaceutical composition for preventing or treating dyslipidemia may be a composition for oral administration.

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

The present invention also provides a pharmaceutical composition for preventing or treating fatty liver comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine and a pharmaceutically acceptable salt thereof.

In the present invention, the pharmaceutically acceptable salt of protamine may be protamine sulfate.

In the present invention, the pharmaceutical composition for preventing or treating fatty liver disease may be a composition for oral administration.

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

The present invention also provides a food composition for preventing or improving dyslipidemia containing ursodeoxycholic acid or a foodologically acceptable salt thereof, and protamine and a foodologically acceptable salt thereof.

In the present invention, the food composition for preventing or improving dyslipidemia may be characterized as having an effect of reducing triglycerides in the blood.

In the present invention, the foodologically acceptable salt of protamine may be protamine sulfate.

In the present invention, the ursodeoxycholic acid or its food-acceptable salt and the protamine and its food-acceptable salt may be mixed to form a nanocomposite.

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

In the present invention, the foodologically acceptable salt of protamine may be protamine sulfate.

In the present invention, the ursodeoxycholic acid or its food-acceptable salt and the protamine and its food-acceptable salt may be mixed to form a nanocomposite.

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

Figure 1 is a TEM photograph 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 administering distilled water, protamine, ursodeoxycholic acid, ursodeoxycholic acid, and protamine mixture to dyslipidemic mice, respectively, and observing body weight changes.
Figure 4 shows the results of administering distilled water, protamine, ursodeoxycholic acid, ursodeoxycholic acid, and protamine mixture to dyslipidemic mice, respectively, and confirming the blood low-density lipoprotein level (LDL).
Figure 5 shows the results of administering distilled water, protamine, ursodeoxycholic acid, ursodeoxycholic acid, and protamine mixture to dyslipidemic mice, respectively, and checking the blood sugar concentration.
Figure 6 shows the results of administering distilled water, protamine, ursodeoxycholic acid, ursodeoxycholic acid, and protamine mixture to dyslipidemic mice, respectively, and checking blood triglyceride levels.

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

In the present invention, it was confirmed that when ursodeoxycholic acid and protamine are used in combination, the effect of reducing neutral lipids in the blood is excellent, and thus it can be effective in preventing, improving, and/or treating dyslipidemia.

Therefore, in one aspect, the present invention relates to a pharmaceutical composition for preventing or treating dyslipidemia comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine and a pharmaceutically acceptable salt thereof. It's about.

In addition, from another aspect, the present invention relates to a pharmaceutical composition for preventing or treating fatty liver comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and protamine and a pharmaceutically acceptable salt thereof. .

In the present invention, the pharmaceutical composition may be characterized as having a blood neutral fat reduction effect, but its effect is not limited thereto.

In the present invention, the pharmaceutically acceptable salt of protamine may be protamine sulfate or chloride (-Cl), but is not limited thereto.

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

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

In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine and a pharmaceutically acceptable salt thereof are used simultaneously or sequentially for the prevention or treatment of dyslipidemia and/or fatty liver. It may be characterized as being administered.

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

In the present invention, the fatty liver refers to non-alcoholic fatty liver disease (NAFLD), simple steatosis in the liver, steatohepatitis, or hepatocirrhosis. ), but is not limited to this.

In the present invention, the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof may be contained in an amount of 0.1 to 75% by weight, preferably 1 to 50% by weight, based on the total weight of the composition, but is limited thereto. It doesn't work.

In the present invention, the protamine and its pharmaceutically acceptable salt may be contained in an amount of 0.1 to 75% by weight, preferably 1 to 50% by weight, based on the total weight of the composition, but is not limited thereto.

In the present invention, the ratio of the ursodeoxycholic acid or its pharmaceutically acceptable salt and the protamine and its pharmaceutically acceptable salt is 1:0.01 to 10, preferably 1:0.1 to 5, more preferably. It may be administered at a weight ratio of 1:0.5 to 2, most preferably 1:1, but is not limited thereto.

As used herein, the term “pharmaceutical composition” or “pharmaceutical formulation” refers to a mixture comprising pharmaceutically acceptable excipients such as diluents or carriers that render the compositions of the present invention particularly suitable for therapeutic use. means. According to some embodiments, a pharmaceutical composition containing the composition of the present invention may be provided by administering a therapeutically effective amount to a subject according to need. In some embodiments, compositions of the present invention can be administered to humans.

As used herein, “effective amount” or “therapeutically-effective amount” refers to the amount of a compound or composition (e.g., a compound or composition of the invention) sufficient to achieve a beneficial or desired result. Refers to an amount. An effective amount may be administered in one or more administrations, applications, or dosages and is not intended to be limited to a particular agent or route of administration.

Although the pharmaceutical compositions provided by the present invention relate in principle to pharmaceutical compositions for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to all types of animals. That is, the pharmaceutical composition according to the present invention is suitable for use in animals requiring veterinary treatment, such as livestock (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.) and laboratory animals. It can also be administered to other mammals such as rats, mice, guinea pigs, etc. A skilled veterinary pharmacologist with a good understanding of the modifications of pharmaceutical compositions for administration to various animals can design and/or perform such modifications, if necessary, simply by routine experimentation.

The pharmaceutical compositions described in the present invention may be prepared by any method known in the field of pharmacology or as discussed later in the text. Generally, these methods for tableting involve the steps of associating the active ingredient with excipients and/or one or more other auxiliary ingredients, followed by shaping and/or packaging the product into the desired single- or multi-dose units, if necessary or desired. Includes steps.

The pharmaceutical compositions 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 a discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of active ingredient is generally equal to the dosage of active ingredient administered to the subject and/or a convenient fraction of such dosage such as, for example, one-half or one-third of the dosage.

The relative amounts of the active ingredient, pharmaceutically acceptable excipients, and/or any additional ingredients in the pharmaceutical compositions of the invention will vary depending on the identity, size, and/or disorder of the subject being treated and the route by which the composition is administered. . By way of example, the composition may include 0.001% to 100% (w/w) active ingredient.

As used herein, a pharmaceutically acceptable excipient is any solvent, dispersion medium, diluent, or other liquid vehicle, dispersion or suspension aid, surface active agent, isotonic agent, thickener or emulsifier suitable for the purpose of the particular dosage form; Contains preservatives, solid binders, lubricants, etc. Remington's (The Science and Practice of Pharmacy, 21st Edition, A. R. Gennaro, (Lippincott, Williams & Wilkins, Baltimore, MD, 2006) describes various excipients used in the preparation of pharmaceutical compositions and known methods for their preparation. The technology is disclosed, except that any conventional carrier medium is incompatible with the substance or derivative thereof, for example by providing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component of the pharmaceutical composition. and their uses are considered to be within the scope of the present invention. Pharmaceutically acceptable excipients are at least 95%, 96%, 97%, 98%, 99%, or 100% pure.

The excipients are approved for human and veterinary use. In some embodiments, the excipient is approved by the Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopeia (USP), European Pharmacopoeia (EP), British Pharmacopoeia, and/or International Pharmacopoeia (EP).

Pharmaceutically acceptable excipients used in the preparation of pharmaceutical compositions include, but are not limited to, inert diluents, dispersants and/or granulating agents, surface active agents and/or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants, and/or oils. Not limited.

Such excipients may optionally be included in the formulations of the present invention. Excipients such as cocoa butter and suppository waxes, colorants, coating agents, sweeteners, flavors, and perfumers may be present in the composition at the discretion of the formulator.

Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dried starch, Including, but not limited to, corn starch, powdered sugar, and combinations thereof.

Exemplary granulating and/or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation-exchange resins. , 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, pregelatinized starch (Starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (vegeum), sodium lauryl sulfate, quaternary ammonium compounds, and these. Including, but not limited to, combinations of.

Exemplary surface active agents 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 veegum [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. carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymers), carrageenans, cellulose derivatives (e.g. carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g. polyoxymethylcellulose) Ethylene Sorbitan Monolaurate [Tween 20], Polyoxyethylene Sorbitan [Tween 60], Polyoxyethylene Sorbitan Monooleate [Tween 80], Sorbitan Monopalmitate [Span 40], Sorbitan Monostearate [ span 60], sorbitan tristearate [span 65], glyceryl monooleate, sorbitan monooleate [span 80]), polyoxyethylene ester (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 ethers, ( For example, polyoxyethylene lauryl ether [Brise 30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl. Including, but not limited to, lauray, sodium lauryl sulfate, pluronic F 68, poloxamer 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and/or combinations thereof. No.

Exemplary binders include starches (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, extract of Irish moss, Farnwer gum, Shatty gum, Isapol Husk's slime, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl) cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (bigum), and lachi arabogalactan); alginate; polyethylene oxide; polyethylene glycol; inorganic calcium salt; silicic acid; polymethacrylate; wax; water; Alcohol; and combinations thereof.

Exemplary preservatives may include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives. Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, bisulfite. Including, but not limited to, sodium sulfate, sodium metabisulfite, and sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, disodium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and trisodium edetate. . Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, Includes, but is not limited to, hexetidine, imidurea, 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 acidic 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 tocopherol, tocopherol acetate, deteroxyme mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluende (BHT), ethylenediamine, sodium lauryl sulfate (SLS), Sodium Lauryl Ether Sulfate (SLES), Sodium Bisulfite, Sodium Metabisulfite, Potassium Sulfite, Potassium Metabisulfite, Glydant Plus, Fenonib, Methylparaben, Low Mol 115, Germaben II, Neolon, Katon, and E Including, but not limited to, Uxil. In certain embodiments, the preservative is an antioxidant. In another embodiment, the preservative is a chelating agent.

Exemplary buffering agents include citrate buffer solution, acetate buffer solution, phosphate buffer solution, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium gluvionate, calcium gluceptate, calcium gluconate, D-gluconic acid, glyceroside. Calcium phosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixture, dibasic potassium phosphate, monobasic Basic 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 -Includes, but is not limited to, free water, isotonic saline, Ringer's solution, ethyl alcohol, and combinations thereof.

Exemplary lubricants include 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. Including, but not limited to, lauryl sulfate, and combinations thereof.

Exemplary oils include almond, apricot kernel, avocado, babassu palm, bergamot, black currant seed, borage, cayenne, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee. , corn, cotton seed, emu, eucalyptus, evening primrose, fish, flax seed, geraniol, pumpkin, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litthea cucumber. beba, macadamia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange-orange rappi, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower. , sandalwood, sasquana, savory, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germ oil. Exemplary oils include butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil. , and combinations thereof.

Liquid dosage forms 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, liquid dosage forms may contain inert diluents such as, for example, water or other solvents, solubilizers and emulsifiers commonly used in the art such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl. Alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed, peanut, corn, sprout, olive, castor, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, It may also include polyethylene glycol and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the pharmaceutical compositions of the invention are mixed with solubilizing agents such as cremophor, alcohols, oils, denatured oils, glycols, polysorbates, cyclodextrins, polymers, and combinations thereof.

Injectable preparations, for example, sterile injectable aqueous or oily suspensions, may be prepared according to the known art using dispersing or wetting agents and suspending agents. The sterile injectable preparation may be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, such as solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. Additionally, sterile, fixed oils are commonly employed as solvents or suspending media. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. Additionally, fatty acids such as oleic acid have been used in the preparation of injectables.

Injectable preparations may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. .

To prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This is achieved by using liquid suspensions of crystalline or amorphous substances with low water solubility. The absorption rate of the drug then depends on the dissolution rate, which may ultimately depend on the crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug is achieved by dissolving or suspending the drug in an oil vehicle.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active ingredient may be combined with one or more inert pharmaceutically acceptable excipients or carriers such as sodium citrate or dicalcium 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 retarders such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) humectants 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, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof.

In the case of capsules, tablets and pills, dosage forms may contain buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using lactose or milk sugar as well as high molecular weight polyethylene glycols and the like as such excipients. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and may be compositions that release the active ingredient only, or preferentially, in a specific part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using lactose or milk sugar as well as high molecular weight polyethylene glycols and the like as such excipients.

The active ingredient may be in micro-encapsulated form with one or more of the excipients described above. Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, controlled release coatings, and other coatings well known in the pharmaceutical formulation art. 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 contain additional substances other than inert diluents as in common practice, such as tablet lubricants and other tablet auxiliaries such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, dosage forms may also contain buffering agents. They may optionally contain opacifying agents and may be compositions that release the active ingredient only, or preferentially, in a specific part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of the pharmaceutical compositions of the present invention may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and/or patches. Generally, the active ingredient is admixed under conditions of sterility with a pharmaceutically acceptable carrier and/or any necessary preservatives and/or buffering agents that may be required. Additionally, the present invention contemplates the use of transdermal patches, which often have the additional advantage of providing controlled delivery of the active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and/or dispersing the active ingredient in a suitable medium. Alternatively or additionally, the rate may be controlled by providing a rate controlling membrane and/or dispersing the active ingredient in a polymer matrix and/or gel.

Formulations for topical administration include, but are not limited to, liquid and/or semi-liquid preparations such as liniments, 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 solubility limit of the active ingredient in the solvent, topically-administrable formulations may also comprise, for example, from about 1% to about 10% (w/w) of the active ingredient. Formulations for topical administration may further comprise one or more additional ingredients described herein.

Pharmaceutical compositions of the invention described herein are typically prepared in dosage unit form for easy administration and uniform administration. However, it will be understood that the total daily dosage of the composition of the present invention will be determined by the attending physician within the scope of sound medical judgment. The particular therapeutically effective dose level for any particular subject will depend on a variety of factors, including the disease, disorder, or disorder being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; The subject's age, weight, general health, gender, and diet; the time of administration, route of administration, and rate of excretion of the particular active ingredient employed; duration of treatment; Drugs used in combination or simultaneously with the specific active ingredients employed; and factors well known in the medical field.

The pharmaceutical composition of the present invention may be administered by any route. In some embodiments, the pharmaceutical composition can be administered orally, intravenously, intramuscularly, intraarterially, intramedullarily, intrathecally, subcutaneously, intracerebroventricularly, transdermally, intradermally, rectally, intravaginally, intraperitoneally, or topically (powder, ointment, cream). , and/or by droplets), mucous membranes, nose, mouth, enteral, sublingual; endotracheal instillation, bronchial instillation, and/or inhalation; and/or administered by various routes, including oral spray, nasal spray, and/or aerosol. Particularly contemplated routes are permeable intravenous injection, local administration via the blood and/or lymphatic supply, and/or direct administration to the affected area. In general, the most appropriate route of administration will depend on a variety of factors, including the properties of the agent (e.g., stability in the environment of the gastrointestinal tract), and the disorder of the subject (e.g., whether the subject can tolerate oral administration). will be.

In certain embodiments, the pharmaceutical compositions of the invention can be administered at a daily dose of about 0.001 mg/kg to about 100 mg/kg, about 0.01 mg/kg to about 50 mg/kg, or about 0.1 mg/kg to about 40 mg/kg of the subject's body weight. 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. The desired therapeutic effect may be obtained by administering the drug at a dosage level sufficient to deliver it more than once a day. The intended dosage may be delivered three times a day, twice a day, daily, every two days, every three days, weekly, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dosage may be delivered via multiple administrations (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or more administrations). .

It will be understood that the dosage ranges described herein provide guidance for administration of pharmaceutical compositions provided to adults. For example, the amount administered to a child or adolescent may be determined by a physician or person skilled in the art, and may be less or the same as that administered to an adult. The exact amount of a peptide according to the invention required to achieve an effective amount will vary from subject to subject, depending, for example, on the subject's species, age, and overall disorder, severity of side effects or disorders, identity of the specific compound, mode of administration, etc.

It will be understood that the pharmaceutical compositions of the present invention can be used in combination therapy. The particular combination of treatments (treatments or procedures) for use in combination therapy will take into account the desired therapeutic effect to be achieved and the suitability of the desired treatments and/or procedures.

The pharmaceutical composition of the present invention can be administered alone or in combination with one or more therapeutically active agents. As for "combination", it is not intended to imply that the agents must be administered at the same time and/or be formulated for delivery together, although the following delivery methods are within the scope of the present invention. The composition may be administered concurrently with, prior to, or subsequent to one or more other therapeutic agents or medical procedures. Generally, each agent will be administered at a dose and/or time schedule established for that agent. Additionally, the present invention provides a pharmaceutical form of the present invention in combination with agents capable of improving its bioavailability, reducing and/or modifying its metabolism, inhibiting its secretion, and/or modifying its distribution within the body. It encompasses delivering the composition. It will be further understood that the pharmaceutical composition and the therapeutically active agent of the invention used in this combination may be administered together in a single composition or may be administered separately in different compositions.

The particular combination used in combination therapy will take into account the desired therapeutic effect to be achieved and/or the suitability of the procedure and/or therapeutically active agent comprising the peptide of the invention. The combination used may achieve the desired effect for the same disorder (e.g., the pharmaceutical composition of the present invention may be administered in combination with another therapeutically active agent (e.g., a second therapeutic agent) used to treat the same disorder. may be), and/or they may achieve different effects (e.g., controlling any side effects).

As used herein, “therapeutically active agent” refers to any substance used as a medicine to treat, prevent, delay, reduce or ameliorate a disorder, and refers to substances used in treatment, including prophylactic and curative treatments. .

In some embodiments, the pharmaceutical compositions of the invention treat, alleviate, ameliorate, alleviate, delay the onset, inhibit the progression, reduce the severity, and/or the incidence of one or more symptoms or characteristics. Can be administered in combination with any therapeutically active agent or procedure (e.g., surgery, radiation therapy) useful for reducing .

In one aspect, the present invention may be provided to a subject suffering from dyslipidemia and/or fatty liver as a kit for treating the disease.

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 include one or more unit dosage forms containing a dosage of the pharmaceutical composition as described herein effective for treating the disease in a subject. In some embodiments, the subject is a human patient.

In some embodiments, the kit further includes 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.

The term “about” used in the present invention may be interpreted to mean approximately, roughly, approximately, or to a certain extent. When the term "about" is used with a numeric range, it is interpreted as modifying that range by extending the boundaries above and below the specified numeric value. Generally, the term "about" is used herein to modify a numerical value above or below a specified value by a variance of 10%.

“Individual,” “patient,” and “subject” are used interchangeably and refer to mammals, such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, and sheep. , horses, or any animal, including primates, including humans.

In the present invention, unless otherwise stated, "treatment" means reversing, alleviating, inhibiting the progression of, or preventing the disease or disease to which the term applies, or one or more symptoms of the disease or disease. means, and the term treatment as used herein refers to the act of treating when “treating” is defined as above. Accordingly, “treatment” or “therapy” of a disease in a mammal may include one or more of the following:

(1) arresting the development of the disease;

(2) preventing the spread of disease;

(3) alleviating disease;

(4) preventing disease recurrence and

(5) Alleviating symptoms of disease (palliating)

In the present invention, unless otherwise specified, “prevention” means all actions that inhibit or delay the occurrence, spread, and recurrence of dyslipidemia or fatty liver by administering the pharmaceutical composition according to the present invention.

From another perspective, the present invention relates to a food composition for preventing or improving dyslipidemia comprising ursodeoxycholic acid or a foodologically acceptable salt thereof, and protamine and a foodologically acceptable salt thereof. .

From another perspective, the present invention relates to a food composition for preventing or improving fatty liver comprising ursodeoxycholic acid or a foodologically acceptable salt thereof, and protamine and a foodologically acceptable salt thereof.

In the present invention, the food composition is characterized in that it exerts an effect of reducing neutral fat in the blood, and its effect is not limited thereto.

In the present invention, the foodologically acceptable salt of protamine may be protamine sulfate, but is not limited thereto.

In the present invention, the ursodeoxycholic acid or its food-acceptable salt and the protamine and its food-acceptable salt may be mixed to form a nanocomposite.

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

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 to this.

In the present invention, the ursodeoxycholic acid or a foodologically acceptable salt thereof may be contained in an amount of 0.1 to 75% by weight, preferably 1 to 50% by weight, based on the total weight of the composition, but is limited thereto. It doesn't work.

In the present invention, the protamine and its food-acceptable salt may be contained in an amount of 0.1 to 75% by weight, preferably 1 to 50% by weight, based on the total weight of the composition, but is not limited thereto.

In the present invention, the ratio of the ursodeoxycholic acid or its foodologically acceptable salt and the protamine and its foodologically acceptable salt is 1:0.01 to 10, preferably 1:0.1 to 5, more preferably. It may be administered at a weight or volume ratio of 1:0.5 to 2, most preferably 1:1, but is not limited thereto.

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

There is no particular limitation to the food as long as it contains the food composition, and it can also be used as an additive to other foods. Examples of foods to which the food composition can be added include meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, and drink preparations. , alcoholic beverages, vitamin complexes, etc., and includes all health foods in the conventional sense.

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

In addition, among the functional foods of the present invention, health drinks may contain various flavoring agents or natural carbohydrates as additional ingredients like ordinary drinks. Flavoring agents include natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame. Natural carbohydrates 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.

Example

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Example 1: Preparation of a mixture of ursodeoxycholic acid and protamine

Ursodeoxycholic acid (Sigma-Aldrich, U5127) is stored at room temperature. During the experiment, 10 mg was weighed and slowly dissolved in 1 mL of DMSO solvent, followed by sonification for about 1 minute to maximize 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 at a volume ratio of 1:10, and distilled water was used as an additional solvent to bring the final concentration to 1 mg/mL. For samples administered to animals, purified water was used as a dilution solution.

Example 2: Analysis of a mixture of ursodeoxycholic acid and protamine

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

The sample solution was sonicated for 1 minute just before measurement, and then 10 uL of the mixture prepared in Example 1 was applied on 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 using the EVO MA 10 (Carl Zeiss) model of a scanning electron microscope.

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

Example 3. Cytotoxicity analysis of ursodeoxycholic acid and protamine mixture

To evaluate the potential cytotoxicity of the ursodeoxycholic acid and protamine mixture, colonocytes (CT26, Korean Cell Line Bank) were cultured in DMEM medium (Sigma-Aldrich) supplemented with 10% FBS at 37°C and 5% CO. Cultured for 1 week under ₂ conditions. Afterwards, the cells were removed using a trypsin-EDTA solution, distributed into 96 well plates at 1×10 ³ cells/well, and 10 or 100 uM of a mixture of ursodeoxycholic acid and protamine was administered. After 24 hours of incubation, according to the EZ-cytox measurement protocol provided by the manufacturer, 10 uL of EZ-Cytox (Dogenbio) solution was added and the absorbance was measured at 450 nm to calculate cytotoxicity.

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

Example 4. Effect of improving blood lipid levels in a mouse model of dyslipidemia disease

A dyslipidemia animal model was created by receiving 4-week-old C57BL/6J mice (Orient Bio, Korea) and replacing the feed with a high-cholesterol diet (Atherogenic Diet, D12336) the next day. The mouse model is an animal experiment model that can pathologically resemble dyslipidemia that occurs in humans through a high-cholesterol diet. After about 10 days of feed administration, administration of a mixture of ursodeoxycholic acid and protamine was started while maintaining the high-cholesterol diet. Ursodeoxycholic acid and protamine mixture (1:1) was administered daily at 20 mg/kg/day (UDCA 10 mg + Protamine 10 mg). As a control group, only purified water was administered, and as a comparison group, a group administered only 10 mg/kg/day protamine or a group administered only 10 mg/kg/day ursodeoxycholic acid was used. Administration was conducted daily orally using an oral sonde for mice, and the administration dose for each mouse was 200㎕.

4-1. weight change

As a result of measuring the change in mouse body weight over 28 days, as shown in Figure 3, there was a significant difference in body weight in all groups administered with protamine alone, ursodeoxycholic acid alone, and ursodeoxycholic acid and protamine mixture group compared to the control group. did not indicate

4-2. Changes in low-density lipoprotein, sugar, and triglyceride levels in the blood

Blood was collected from mice on the 28th day of administration. The collected blood was left at room temperature for 1 hour, then centrifuged at 3000 rpm for 20 minutes to obtain serum. Afterwards, a blood test was conducted at a testing agency (KP&T) to check changes in low-density lipoprotein, sugar, and neutral fat levels in the blood.

In the case of low-density lipoprotein in the blood, as shown in Figure 4, the low-density lipoprotein (LDL) level was specifically increased in the protamine-only group compared to the control group, and in the ursodeoxycholic-only group and the ursodeoxycholic acid group. And the protamine mixture administration group showed no significant difference in low-density lipoprotein levels compared to the control group.

As a result of measuring blood sugar concentration, as shown in Figure 5, the group administered the ursodeoxycholic acid and protamine mixture showed slightly lower values compared to the group administered distilled water, the group administered only protamine, and the group administered only ursodeoxycholic.

Meanwhile, as a result of serum triglyceride analysis, as shown in Figure 6, it was confirmed that the blood triglyceride level was significantly reduced in the group administered the ursodeoxycholic acid and protamine mixture compared to the group administered distilled water, the group administered only protamine, and the group administered only ursodeoxycholic. did.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (16)

A pharmaceutical composition for preventing or treating dyslipidemia comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof, and protamine and a pharmaceutically acceptable salt thereof.
The pharmaceutical composition according to claim 1, wherein the composition exhibits a blood neutral fat reducing effect.
The pharmaceutical composition according to claim 1, wherein the pharmaceutically acceptable salt of protamine is protamine sulfate.
The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is a composition for oral administration.
The pharmaceutical method of claim 1, wherein the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine and a pharmaceutically acceptable salt thereof are mixed to form a nanocomposite. Composition.
A pharmaceutical composition for preventing or treating fatty liver comprising ursodeoxycholic acid or a pharmaceutically acceptable salt thereof, and protamine and a pharmaceutically acceptable salt thereof.
The pharmaceutical composition according to claim 6, wherein the pharmaceutically acceptable salt of protamine is protamine sulfate.
The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition is a composition for oral administration.
The pharmaceutical method of claim 6, wherein the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof and the protamine and a pharmaceutically acceptable salt thereof are mixed to form a nanocomposite. Composition.
A food composition for preventing or improving dyslipidemia comprising ursodeoxycholic acid or a foodologically acceptable salt thereof, and protamine and a foodologically acceptable salt thereof.
The food composition according to claim 10, wherein the composition exhibits a blood neutral fat reducing effect.
The food composition according to claim 1, wherein the foodologically acceptable salt of protamine is protamine sulfate.
The food composition according to claim 10, wherein the ursodeoxycholic acid or a foodologically acceptable salt thereof and the protamine and a foodologically acceptable salt thereof are mixed to form a nanocomposite. .
A food composition for preventing or improving fatty liver comprising ursodeoxycholic acid or a foodologically acceptable salt thereof, and protamine and a foodologically acceptable salt thereof.
The food composition according to claim 14, wherein the foodologically acceptable salt of protamine is protamine sulfate.
The food composition according to claim 14, wherein the ursodeoxycholic acid or a foodologically acceptable salt thereof and the protamine and a foodologically acceptable salt thereof are mixed to form a nanocomposite. .