Patent Description:
Fats are derived from the accumulation of overfed energy in white adipose tissue and the excessive accumulation thereof in white adipose tissue is commonly referred to as obesity. The breakdown of triglycerides into free fatty acid (FFA) and glycerol, e.g., by hormone sensitive lipase (HSL), is referred to as lipolysis.

For improving obesity including localized fat deposits (LFD), a surgical operation, liposuction, is being carried out. Liposuction, which is also known as lipoplasty or liposculpture suction lipectomy, is a cosmetic surgical operation for removing fats from various sites in the body, such as abdomen, thighs, hips, neck, upper arms, and so on. However, there are concerns that liposuction may cause serious side effects, including wounds, swelling, paralysis and burning sensation, a risk of infection in the surgical site; damage to skin or nerves; and puncture wounds in critical organs. In addition, it requires a significant period of time for treatment and recovery; and also there are risks associated with anesthesia because it requires local anesthesia or general anesthesia during the surgical procedure.

Phosphatidylcholines (PC) are a class of phospholipids that incorporate choline as a headgroup. They are widely found in animals, plants, yeast, and fungi and are also referred to as lecithin or unsaturated lecithin. They are mainly contained in the brain, nerves, blood cells, egg yolk and the like in mammals; and in soybeans, sunflower seeds, wheat germ and the like in plants. Because phosphatidylcholine has four double bonds in its molecule, it is easily oxidized during the preparation or during the storage, thereby the structure thereof being deformed. In order to overcome such disadvantage, the saturated lecithin forms have been developed which are prepared by adding hydrogen to the unsaturated lecithin. US Laid-open Patent Publication No. <CIT> has disclosed a method for reducing adipose tissue which includes injecting lecithin. In addition, US Laid-open Patent Publication No. <CIT> has disclosed a method for reducing localized adipose tissue comprising topically administering to a site at or proximate adipose tissue a composition comprising lecithin and bile acid or a salt thereof. However, the topical injection of lecithin for inducing lipolysis leads to side effect problems such as erythema, inflammation, tissue necrosis, edema, dimpling and the like. And also, when a lecithin-containing formulation is stored for a long period of time, it leads to problems such as lowered stability and lipolytic activity. <CIT> discloses a composition comprising a phospholipid (e.g. dimyristoylphosphatidylglycerol or dipalmitoylphosphatidylglycerol) and/or a bile acid selected from deoxycholic acid, cholic acid, lithocholic acid, chenodeoxycholic acid, hyodeoxycholic acid, trihydroxycoprostanic acid, ursodeoxycholic acid, taurocholic acid and glycocholic acid, in a weight ratio of <NUM>:<NUM> to <NUM> :<NUM>, for use in the pharmaceutical and cosmetic removal of subcutaneous accumulation of fats.

<CIT> discloses an aqueous composition suitable for reducing subcutaneous fat deposits comprising, by weight, about <NUM>% to about <NUM>% phosphatidylcholine (i.e. <NUM>-palmitoyl-<NUM>-oleoylsn-glycero-<NUM>-phosphocholine), a bile acid or bile acid salt selected from deoxycholic acid, cholic acid, lithocholic acid, Na taurocholate, Na taurochenodeoxycholate, Na taurodeoxycholate or glycodeoxycholatewherein the ratio of phosphatidylcholine to bile acid or bile acid salt is between about <NUM> w/w and <NUM> w/w, and a thickening agent.

Subject-matter disclosed herein and not falling within the subject-matter of the claims is intended to be mentioned merely for descriptive purpose.

The present inventors carried out various researches in order to develop a composition for lipolysis or for ameliorating, inhibiting or treating localized fat deposits, which has an excellent lipolytic activity and stability without significant side effects. The present inventors evaluated activities, side effects, and stabilities of various choline derivatives. As the results thereof, the present inventors have found that the compositions containing certain phosphocholine derivatives have a superior and uniform lipolytic activity and stability and can minimize side effects at the administrated or applied site such as inflammation, tis necrosis, etc., in comparison to a conventional phosphatidylcholine-containing formulation.

Therefore, the present invention provides a composition comprising <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof for use in ameliorating, inhibiting or treating obesity as set out in claim <NUM>. And also, the present invention provides a composition as set out in claim <NUM>. The preferred features are as defined in claims <NUM> to <NUM>.

Disclosed herein is a method for inducing lipolysis or for ameliorating, inhibiting or treating localized fat deposits in a mammal in need thereof, comprising administering to the mammal an effective amount of said phosphocholine derivatives.

Disclosed herein is a use of said phosphocholine derivatives for the manufacture of a medicament for inducing lipolysis or for ameliorating, inhibiting or treating localized fat deposits.

In accordance with an aspect of the present invention, there is provided a composition for use in. ameliorating, inhibiting or treating obesity , comprising an effective amount of <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof, wherein the composition is in the form of a pharmaceutical composition for topical administration or in the form of a cosmetic composition for applying to a skin, subcutaneous tissues, muscular tissues, or abdominal tissues.

The composition of the present invention may further comprise an effective amount of bile acid or pharmaceutically acceptable salt thereof. The bile acid or pharmaceutically acceptable salt thereof may be one or more selected from the group consisting of cholic acid, glycocholic acid, glycodeoxycholic acid, deoxycholic acid, taurocholic acid, ursodeoxycholic acid, tauroursodeoxycholic acid, taurodeoxycholic acid, chenodeoxycholic acid, glycoursodeoxycholic acid, sodium deoxycholate, and sodium taurocholate. Preferably, the bile acid or pharmaceutically acceptable salt thereof may be one or more selected from the group consisting of glycocholic acid, deoxycholic acid, taurocholic acid, tauroursodeoxycholic acid, glycoursodeoxycholic acid, sodium deoxycholate, and sodium taurocholate. In an embodiment, a weight ratio of <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof and the bile acid or pharmaceutically acceptable salt thereof may range from <NUM> : <NUM> to <NUM>:<NUM>.

The composition of the present invention is in the form of a pharmaceutical composition for topical administration. The pharmaceutical composition for topical administration may be in the form of a formulation for transdermal administration, in the form of a formulation for subcutaneous administration, in the form of a formulation for intramuscular administration, or in the form of a formulation for intraperitoneal administration. And also, the pharmaceutical composition for topical administration may be in the form of a liquid formulation or in the form of a dry powder formulation; preferably in the form of a solution, an emulsion, or a lyophilized powder. The pharmaceutical composition for topical administration may comprise one or more pharmaceutically acceptable excipients selected from the group consisting of a pH controlling agent, an isotonic agent, a surfactant, a stabilizer, a preservative, a chelating agent, a buffer, and a cryoprotectant; and one or more pharmaceutically acceptable carriers selected from the group consisting of an oil, an organic solvent, and an aqueous solvent.

The composition of the present invention is in the form of a cosmetic composition for applying to a skin, subcutaneous tissues, muscular tissues, or abdominal tissues. The cosmetic composition may be in the form of a liquid or in the form of a dry powder; preferably in the form of a solution, an emulsion, or a lyophilized powder. The cosmetic composition may comprise one or more excipients selected from the group consisting of a pH controlling agent, an isotonic agent, a surfactant, a stabilizer, a preservative, a chelating agent, a buffer, and a cryoprotectant; and one or more carriers selected from the group consisting of an oil, an organic solvent, and an aqueous solvent.

Disclosed herein is a method for inducing lipolysis or for ameliorating, inhibiting or treating localized fat deposits in a mammal in need thereof, comprising administering to the mammal an effective amount of one or more compounds selected from the group consisting of:.

Preferably, in the method of the present disclosure, the compound may be one or more selected from the group consisting of:.

More preferably, in the method of the present disclosure, the compound may be one or more selected from the group consisting of:.

In accordance with the present invention, there is provided a composition comprising <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof for use in ameliorating, inhibiting or treating obesity, wherein the composition is in the form of a pharmaceutical composition for topical administration or in the form of a cosmetic composition for applying to a skin, subcutaneous tissues, muscular tissues, or abdominal tissues.

The composition of the present invention comprising certain phosphocholine derivatives, i.e., <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof shows not only an excellent lipolytic activity but also uniformity in its lipolytic activity in comparison to a conventional phosphatidylcholine-containing formulation. And also, the composition of the present invention can minimize side effects such as inflammation, tissue necrosis, etc., at the administrated or applied site. In addition, the composition of the present invention has excellent storage stability. Accordingly, the composition of the present invention can inhibit non-uniform arrangement of subcutaneous fats and aesthetic damages derived therefrom; and therefore can be usefully applied for inducing lipolysis or for ameliorating, inhibiting or treating localized fat deposits (LFD).

The phosphocholine derivatives according to the present invention have an excellent and uniform lipolytic activity and can minimize side effects such as inflammation, tissue necrosis, etc., at the administered or applied site; and therefore can be usefully applied for inducing lipolysis or for ameliorating, inhibiting or treating localized fat deposits (LFD). Therefore, the present invention provides a composition for use in ameliorating, inhibiting or treating obesity comprising <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof as an active ingredient. Disclosed herein is a composition for inducing lipolysis or for ameliorating, inhibiting or treating localized fat deposits, comprising an effective amount of a compound of Formula <NUM> or pharmaceutically acceptable salt thereof as an active ingredient:
<CHM>
wherein,
X and Y are, independently of each other, a C<NUM>~C<NUM> alkyl group; or a C<NUM>~C<NUM> alkenyl group having one or two double bonds, with the proviso that X and Y are not a C<NUM> alkyl group at the same time; and X and Y are not a C<NUM> alkenyl group having two double bonds at the same time.

In the composition of the present disclosure, the compound of Formula <NUM> may be one or more selected from the group consisting of:.

Preferably, in the composition of the present disclosure, the compound of Formula <NUM> may be one or more selected from the group consisting of:.

More preferably, in the composition of the present disclosure, the compound of Formula <NUM> may be one or more selected from the group consisting of:.

In the composition of the present invention, the compound of Formula <NUM> is <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine (DMPC).

The compounds of Formula <NUM> or salt thereof, which are known compounds, may be prepared according to known methods. For example, the compound of Formula <NUM> or salt thereof may be prepared by hydrolyzing and purifying soy bean lecithin to obtain glycero phosphocholine; and then reacting the glycero phosphocholine with a fatty acid such as myristic acid. And also, the compound of Formula <NUM> or salt thereof can be purchased as it is commercially available (for example, Sigma-Aldrich).

The compound of Formula <NUM> or pharmaceutically acceptable salt thereof may have substituents containing asymmetric carbon and therefore be in the form of racemic mixture (RS) or in forms of optical isomers, such as (R) or (S) isomer. Therefore, the compound of Formula <NUM> or pharmaceutically acceptable salt thereof comprises both racemic mixture (RS) and optical isomers such as (R) or (S) isomer unless described otherwise.

The compound of Formula <NUM> of the present invention may be in a pharmaceutically acceptable salt form. The salt may be an acid addition salt form, which includes e.g., salts derived from an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, or carbonic acid; and salts derived from an organic acid such as citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, lactobionic acid, salicylic acid, malonic acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, p-toluenesulfonic acid, glutamic acid, or aspartic acid, but not limited thereto. And also, the salt includes an alkali metal salt such as lithium salt, sodium salt, or potassium salt; an alkaline earth metal salt such as calcium salt or magnesium salt; or a chromium salt.

The composition of the present invention further comprise an effective amount of bile acid or pharmaceutically acceptable salt thereof as an active ingredient, in addition to <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof. The bile acid may be one or more selected from the group consisting of cholic acid, glycocholic acid, glycodeoxycholic acid, deoxycholic acid, taurocholic acid, ursodeoxycholic acid, tauroursodeoxycholic acid, taurodeoxycholic acid, chenodeoxycholic acid, glycoursodeoxycholic acid, sodium deoxycholate, and sodium taurocholate. Preferably, the bile acid or pharmaceutically acceptable salt thereof may be one or more selected from the group consisting of glycocholic acid, deoxycholic acid, taurocholic acid, tauroursodeoxycholic acid, glycoursodeoxycholic acid, sodium deoxycholate, and sodium taurocholate. More preferably, the bile acid or pharmaceutically acceptable salt thereof may be one or more selected from the group consisting of glycocholic acid, deoxycholic acid, tauroursodeoxycholic acid, sodium deoxycholate, and sodium taurocholate. Most preferably, the bile acid or pharmaceutically acceptable salt thereof may be glycocholic acid or its salt (e.g., sodium salt) or taurocholic acid or its salt (e.g., sodium salt). When the composition of the present invention further comprises an effective amount of bile acid or pharmaceutically acceptable salt thereof as an active ingredient, in addition to the compound of Formula <NUM> or pharmaceutically acceptable salt thereof, a weight ratio of the compound of Formula <NUM> or pharmaceutically acceptable salt thereof and the bile acid or pharmaceutically acceptable salt thereof may range from <NUM> : <NUM> to <NUM> : <NUM>, preferably from <NUM> : <NUM> to <NUM> : <NUM>, more preferably from <NUM> : <NUM> to <NUM> : <NUM>, still more preferably range from <NUM> : <NUM> to <NUM> : <NUM>, in order to minimize side effects such as inflammation, tissue necrosis, etc. and to provide effective lipolysis.

The composition of the present invention may be administered parenterally, more preferably, topically administered through parenteral route. The topical administration includes topical application(s) to the regions such as under-eye, under-chin, under-arm, hip, calf, back, thigh, ankle, abdomen, and the like. For example, the composition of the present invention is in the form of a pharmaceutical composition for topical administration. The pharmaceutical composition for topical administration may be in the form of a formulation for transdermal administration, in the form of subcutaneous administration, in the form of intramuscular administration, or in the form of intraperitoneal administration. And also, the pharmaceutical composition for topical administration may be a formulation for single administration or multiple administrations. The formulation for multiple administrations may be prepared to be suitable for administering in a volume of about <NUM> at least once a day at intervals of <NUM> to <NUM> days. If necessary, the formulation for multiple administrations may be repeatedly administered to separated parts at a distance of <NUM> to <NUM>.

The pharmaceutical composition for topical administration may be in the form of a liquid formulation or in the form of a dry powder formulation. The liquid formulation includes a solution, an emulsion, a suspension, and the like, preferably in the form of solutions or emulsions. The liquid formulation may be sterile-filtered with a bacterial filter or the like and then filled into an ampule or a vial. The dry powder formulation includes a power form obtained by drying a solution, an emulsion, a suspension and the like through conventional drying methods such as rotary evaporation drying, spray drying, fluidized bed drying, and freeze drying (lyophilization), preferably through freeze drying. The dry powder formulation may be diluted with water for injection, sterile distilled water, water for injection, physiological saline, a glucose solution, a glucose injection, a xylitol injection, a D-mannitol injection, a fructose injection, a Dexran-<NUM> injection, a Dexran-<NUM> injection, an amino acid injection, a Ringer's solution, a lactated Ringer's solution, and the like, before administering to a subject.

The pharmaceutical composition for topical administration in the form of a liquid formulation or in the form of a dry powder formulation may be prepared according to the conventional methods used in the field of pharmaceutics, using pharmaceutically acceptable excipients and/or carriers. Therefore, the pharmaceutical composition for topical administration may comprise one or more pharmaceutically acceptable excipients selected from the group consisting of a pH controlling agent, an isotonic agent, a surfactant, a stabilizer, a preservative (or an antimicrobial agent), a chelating agent, a buffer, and a cryoprotectant; and one or more pharmaceutically acceptable carriers selected from the group consisting of an oil, an organic solvent, and an aqueous solvent.

The pH controlling agent includes a pH controlling agent conventionally used in a formulation for injection, for example, sodium hydroxide, citric acid, acetic acid, phosphoric acid, gluconic acid, ascorbic acid, succinic acid and the lie, but not limited thereto. The pharmaceutical composition of the present invention may have a pH ranging from pH <NUM> to pH <NUM>, preferably a pH ranging from pH <NUM> to pH <NUM>. The pharmaceutical composition having said pH range may minimize pain and/or inflammation when it is topically administered.

The isotonic agent includes a sugar, a sugar alcohol, a salt, and the like, for example glucose, glycerin, sodium chloride, calcium chloride, sodium sulfate, glycerin, propylene glycol, polyethylene glycol (e.g., polyethylene glycol having <NUM> or less of molecular weight), dextrose, hydroxypropyl betadex, mannitol, potassium chloride, dextran, ficoll, gelatin, hydroxyethyl starch, and the like, but not limited thereto. In an embodiment, the isotonic agent may be glycerin and/or sodium chloride. Especially, glycerin may also help improve stability of the formulation. The isotonic agent may be used in an amount suitable for providing a physiologically acceptable osmolality.

The surfactant includes ionic, non-ionic, and/or amphoteric surfactants. In an embodiment, the surfactant may be a non-ionic surfactant. The non-ionic surfactant includes for example polyoxyethylene sorbitan fatty acid ester (e.g., Tween series surfactants), polyoxyethylene polyoxypropylene block copolymer (e.g., Poloxamer series surfactants), and the like, but not limited thereto.

The stabilizer includes for example cholesterol, β-cholesterol, sitosterol, ergosterol, stigmasterol, stigmasterol acetate, lanosterol, and a combination thereof, but not limited thereto. In an embodiment, the stabilizer may be cholesterol.

The preservative includes antimicrobial agents conventionally used in the field of pharmaceutics. The preservative includes benzyl alcohol, glycerin, m-cresol, phenol, benzalkonium chloride, benzethonium chloride, acacia, albumin, alcohol, alginic acid, ascorbyl palmitate, aspartame, boric acid, citric acid, glycerin, pentetic acid, sodium acetate, sorbic acid, chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid, cholesterol, and the like, but not limited thereto. In an embodiment, the preservative may be benzyl alcohol and/or glycerin.

The chelating agent includes ethylenediaminetetraacetic acid (EDTA), buthylenediaminetetraacetic acid, cyclohexane-<NUM>,<NUM>-diaminetetraacetic acid (CyDTA), diethylenetriaminepentaacetic acid (DETPA), ethylenediaminetetrapropionic acid, (hydroxyethyl)ethylenediaminetriacetic acid (HEDTA;), ethylenediaminetetra(methylenephosphonic acid (EDTMP), triethylenetetraminehexaacetic acid (TTHA), <NUM>,<NUM>-diamino-<NUM>-hydroxypropane-N,N,N',N'-tetracetic acid (DHPTA), methyliminodiacetic acid, propylenediaminetetracetic acid, <NUM>,<NUM>,<NUM>-triazacyclodecane-N,N',N"-tris(methylenephosphonic acid (DOTRP), <NUM>,<NUM>,<NUM>,<NUM>-tetraazacyclodecane-N,N',N",N"'-tetrakis(methylenephosphonic acid (DOTP), nitrilotris(methylene)triphosphonic acid, diethylenetriaminephenta(methylenephosphonic acid (DETAP), aminotri(methylenephosphonic acid, <NUM>-hydroxyethylene-<NUM>,<NUM>-diphosphonic acid, bis(hexamethylene)triaminephosphonic acid, <NUM>,<NUM>,<NUM>-triazacyclononan-N,N',N"-tris(methylenephosphonic acid (NOTP), <NUM>-phophonobuthan-<NUM>,<NUM>,<NUM>-tricarboxylic acid, nitrilotriacetic acid (NTA), citric acid, fumaric acid, malic acid, maltol, tartaric acid, gluconic acid, glyceric acid, oxalic acid, phthalic acid, maleic acid, mandelic acid, malonic acid, lactic acid, salicylic acid, methyl salicylate, <NUM>-sulfosalicylic acid, gallic acid, propyl gallate, pyrogallol, <NUM>-hydroxyquinoline, cysteine, and the like, but not limited thereto.

The buffer includes disodium hydrogen phosphate (dibasic sodium phosphate), citric acid, sodium citrate hydrate, potassium citrate, acetic acid, sodium acetate, sodium carbonate, calcium carbonate, tricalcium phosphate, calcium lactate, glycine, maleic acid, malic acid, sodium glutamate, monosodium glutamate, sodium lactate, sodium phosphate, and the mixture thereof, but not limited thereto.

The cryoprotectant may be a sugar, a sugar alcohol, or a mixture thereof. The sugar may be one or more selected from the group consisting of lactose, maltose, sucrose, mannose, trehalose, xylose, fructose, and raffinose. The sugar alcohol may be one or more selected from the group consisting of mannitol, sorbitol, inositol, maltitol, xylitol, and lactitol. And also, the cryoprotectant may additionally include glycine, histidine, polyvinylpyrrolidone (PVP), and the like, but not limited thereto.

The organic solvent includes an alcohol such as ethanol (including anhydrous ethanol, fermented spirits and the like), methanol, acetone, di-acetone, octanol, isopropyl alcohol, lauryl alcohol, polyvinyl alcohol; and a glycol such as polyethylene glycol, ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, pentylene glycol, thioglycolic acid, but not limited thereto.

The oil includes vegetable oils, medium chain triglycerides (MCTs), cholesterol, glyceryl stearate, oleic acid, and the like, but not limited thereto. In an embodiment, the oil may be a vegetable oil and/or oleic acid. The vegetable oil may be selected from the group consisting of cottonseed oil, corn oil, sesame oil, soybean oil, olive oil, coconut oil, peanut oil, sunflower oil, safflower oil, almond oil, avocado oil, palm oil, palm kernel oil, babassu oil, beech nut oil, linseed oil, canola oil, and the combination thereof. In an embodiment, the vegetable oil may be soybean oil.

The aqueous solvent includes water for injection, sterile distilled water, saline, an aqueous dextrose solution, and an aqueous sucrose solution, and so on without limitation.

The pharmaceutical composition for topical administration according to the present invention is typically contained in a sealed and sterilized plastic or organic container. The container may be provided in the form of a defined volume such as an ampoule, a vial, a syringe or a cartridge, or may be provided in the form of a large volume such as a bag for injection or a bottle for injection.

In the pharmaceutical composition for topical administration according to the present invention, the compound of Formula <NUM> or pharmaceutically acceptable salt thereof may be administered in an effective amount ranging from about <NUM>/kg to about <NUM>,<NUM>/kg per day, which may be changed according to patient's age, body weight, susceptibility, symptoms, dosage form, and the like. In an embodiment, the compound of Formula <NUM> or pharmaceutically acceptable salt thereof may be included in an amount ranging from <NUM> to <NUM>,<NUM>, preferably in an amount ranging from <NUM> to <NUM>, per unit formulation. And also, as mentioned in the above, the effective amount of said bile acid or a pharmaceutically acceptable salt thereof may be appropriately determined through considering the weight ratio with the compound of Formula <NUM> or pharmaceutically acceptable salt thereof.

The composition of the present invention may be also in the form of a cosmetic composition for applying to a skin, subcutaneous tissues, muscular tissues, or abdominal tissues. The cosmetic composition of the present invention may be prepared to be suitable for applying in a volume of about <NUM> at least once a day at intervals of <NUM> to <NUM> days. If necessary, the cosmetic composition of the present invention may be repeatedly applied to separated parts at a distance of <NUM> to <NUM>.

The cosmetic composition of the present invention may be in various forms and the forms are not limited. That is, the cosmetic composition of the present invention may be in conventional cosmetic composition forms such as cream, pack, lotion, essence, cleansing water, foundation, makeup base, topical injection and the like. The cosmetic composition may be in the form of a liquid or in the form of a dry powder. The liquid form includes a solution, an emulsion, a suspension, and the like, preferably in the form of solutions or emulsions. If necessary, the liquid form may be sterile-filtered with a bacterial filter or the like and then filled into an ampule or a vial. The dry powder form includes a power form obtained by drying a solution, an emulsion, a suspension and the like through conventional drying methods such as rotary evaporation drying, spray drying, fluidized bed drying, and freeze drying (lyophilization), preferably through freeze drying. The dry powder form may be diluted with water for injection, sterile distilled water, water for injection, physiological saline, and the like, before applying to a subject.

The cosmetic composition in the form of a liquid or in the form of a dry powder may be prepared according to the conventional methods used in the field of cosmetics, using excipients and/or carriers. Therefore, the cosmetic composition may comprise one or more excipients selected from the group consisting of a pH controlling agent, an isotonic agent, a surfactant, a stabilizer, a preservative, a chelating agent, a buffer, and a cryoprotectant; and one or more carriers selected from the group consisting of an oil, an organic solvent, and an aqueous solvent. Said excipients and carriers are the same as in described in the above. And also, the cosmetic composition is typically contained in a sealed and sterilized plastic or organic container. The container may be provided in the form of a defined volume such as an ampoule, a vial, a syringe or a cartridge, or may be provided in the form of a large volume such as a bag for injection or a bottle for injection.

In the cosmetic composition according to the present invention, the compound of Formula <NUM> or pharmaceutically acceptable salt thereof may be applied in an amount ranging from about <NUM>/kg to about <NUM>,<NUM>/kg per day, which may be changed according to subject's age, body weight, susceptibility, symptoms, dosage form, and the like. In an embodiment, the compound of Formula <NUM> or pharmaceutically acceptable salt thereof may be included in an amount ranging from <NUM> to <NUM>,<NUM>, preferably in an amount ranging from <NUM> to <NUM>, per unit cosmetic composition. And also, as mentioned in the above, the amount of said bile acid or a pharmaceutically acceptable salt thereof may be appropriately determined through considering the weight ratio with the compound of Formula <NUM> or pharmaceutically acceptable salt thereof.

The present invention will be described in further detail with reference to the following examples and experimental examples. These examples and experimental examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

The lyophilized formulation in the form of lyophilized dry powder was prepared according to the components and amounts shown in Table <NUM>. The amounts of Table <NUM> represent the weight (mg) of each component. Oleic acid, glycerin, cholesterol, and benzyl alcohol were dissolved in ethanol (<NUM>) under stirring at <NUM> rpm. DMPC was dissolved in the resulting solution under heating at <NUM>±<NUM> to obtain a first solution.

The first solution was added to a solution of mannitol (<NUM>) in water for injection (<NUM>), followed by homogenizing with a high pressure homogenizer (Homogenizer Unidrive x1000D display, CAT GmbH, German) at <NUM>,<NUM> rpm for about <NUM> minutes to obtain an emulsion. The resulting emulsion was subject to sterile filtration with a membrane filter (PVDF <NUM> filter, Millipore, USA) and then lyophilized under the following conditions to obtain a formulation in the form of lyophilized dry powder. The lyophilized formulation was stored in a <NUM> vial.

The lyophilized formulations in the form of lyophilized dry powder were prepared according to the components and amounts shown in Tables <NUM> and <NUM>. The amounts of Tables <NUM> and <NUM> represent the weight (mg) of each component. The first solutions were prepared in accordance with the same procedures as in Example <NUM>, using DMPC or the other phosphocholine derivatives.

In separate vessels, disodium hydrogen phosphate, sodium chloride and sodium hydroxide were dissolved in water for injection (<NUM>) under stirring at <NUM> rpm. In the resulting solution, was the bile acid or a salt thereof dissolved to obtain the respective second solutions.

The first solution (<NUM>) and the second solution (<NUM>) were mixed with each other under heating at <NUM>±<NUM>, followed by homogenizing with a high pressure homogenizer (Homogenizer Unidrive x1000D display, CAT GmbH, German) at <NUM>,<NUM> rpm for about <NUM> minutes to obtain the respective emulsions. The resulting emulsions were added to a solution of mannitol (<NUM>) in water for injection (<NUM>) and then homogenized with a high pressure homogenizer (Homogenizer Unidrive x1000D display, CAT GmbH, German) at <NUM>,<NUM> rpm for about <NUM> minutes to obtain the respective emulsions. The resulting emulsions were subject to sterile filtration with a membrane filter (PVDF <NUM> filter, Millipore, USA) and then lyophilized under the same conditions as in Example <NUM> to obtain formulations in the form of lyophilized dry powder. The lyophilized formulations were stored in a <NUM> vial.

The formulation in the form of solution was prepared according to the components and amounts shown in Table <NUM>. The amounts of Table <NUM> represent the weight (mg) of each component. Benzyl alcohol and Tween <NUM> were dissolved in water for injection (<NUM>-<NUM>) heated to about <NUM>. DMPC was dissolved in the resulting solution under stirring at <NUM> rpm for <NUM> hour. After adjusting the pH by dissolving disodium hydrogen phosphate, sodium chloride, and sodium hydroxide in the resulting solution under heating at about <NUM>, the final volume of the solution was adjusted to <NUM> with water for injection. The resulting solution was subject to sterile filtration with a membrane filter (PVDF <NUM> filter, Millipore, USA) and then filled into a vial.

The formulations in the form of solution were prepared according to the components and amounts shown in Tables <NUM> and <NUM>. The amounts of Tables <NUM> and <NUM> represent the weight (mg) of each component. Benzyl alcohol and Tween <NUM> were dissolved in water for injection (<NUM>-<NUM>) which had been heated to about <NUM>. Each bile acid or a salt thereof was dissolved in the solution under stirring at <NUM> rpm for <NUM> hour (the second solutions). DMPC or the other phosphocholine derivative was dissolved in each second solution under stirring at <NUM> rpm for <NUM> hour, while heating the second solution at about <NUM>. After adjusting the pH by dissolving disodium hydrogen phosphate, sodium chloride, and sodium hydroxide, the final volume of each solution was adjusted to <NUM> with water for injection. The resulting each solution was subject to sterile filtration with a membrane filter (PVDF <NUM> filter, Millipore, USA) and then filled into a vial.

The solution containing phosphatidylcholine was prepared in accordance with the same procedures as in Example <NUM>, except that phosphatidylcholine (<NUM>) was used instead of DMPC.

The solution containing phosphatidylcholine and sodium deoxycholate was prepared in accordance with the same procedures as in Example <NUM>, except that phosphatidylcholine (<NUM>) and sodium deoxycholate (<NUM>) were used instead of DMPC and sodium taurocholate, respectively.

Sprague-Dawley rats (<NUM> weeks old, Orient Bio Inc. (Seongnam, Republic of Korea)) were fed a high-fat diet (D12492, <NUM>% calories from fats, <NUM>% calories from proteins, and <NUM>% calories from carbohydrates, RESEARCH DIET Inc. , New Brunswick, NJ, USA) for <NUM> weeks. The rats were maintained at <NUM>~<NUM> under the condition of <NUM>∼<NUM>% relative humidity with a <NUM> hour light/dark cycle and water was offered ad libitum. The weights of the rats were monitored so as to obtain high-fat rats (fed for <NUM> weeks, <NUM> weeks old in total). The average weights of the rats in the respective groups (<NUM> weeks old in total, n=<NUM>) are shown in Table <NUM>.

Each <NUM> of the vehicle containing no active ingredient, the formulations of Examples <NUM> to <NUM>, and the formulations of Comparative Examples <NUM> and <NUM> was subcutaneously injected into the left groin of the high-fat rats of the respective groups (n=<NUM>) once a week for <NUM> weeks. The formulations of Examples <NUM> to <NUM> were subcutaneously injected after dissolving each lyophilized powder in water for injection (<NUM>).

After <NUM> weeks of recovery from the administration for <NUM> weeks, the rats in each group were sacrificed, followed by observation for inflammation and necrosis; measurement of fat reduction rate at the administration site; evaluation of side effects such as inflammation through Hematoxylin and Eosin staining; and measurement of lipolytic effects through Oil Red O reaction.

After <NUM> weeks of recovery from the administration for <NUM> weeks, the rats in each group were sacrificed and then the subcutaneous injection sites were observed with the naked eyes and tested tactilely. As the results thereof, in the groups administered with the formulations of the present invention, neither necrosis nor inflammation was observed at the injection sites; and also no firm part was touched in the tactile test. However, in the groups administered with the formulations of Comparative Examples <NUM> and <NUM>, necrosis and inflammation were observed at the injection sites with the naked eyes; and also severely firm changes were touched in the tactile test.

After <NUM> weeks of recovery from the administration for <NUM> weeks, the rats in each group were sacrificed and the tissue autopsy thereof was performed. The subcutaneous tissues derived both from the right groin and from the left groin (the administration site) were observed with the naked eyes so as to confirm any abnormal findings; and the photographs thereof were taken. And then, the tissues were taken therefrom and weighed. The relative weight and the reduction rate were calculated according to the following formula. The results thereof are shown in Table <NUM> below and the photographs thereof are shown in <FIG>.

As shown in Table <NUM>, it can be confirmed that the groups administered with the formulations of the present invention showed remarkable decreases in both the organ weight and the relative weight of the administration site, thereby exhibiting excellent lipolytic activities. And also, as shown in <FIG> and <FIG>, it can be confirmed that the lipolytic activities at the administration site (dark circle) are remarkably superior as compared with those at the non-administration site (dotted circle), when observed with the naked eye.

After the abdomen and the groin of the sacrificed rats were dissected, each subcutaneous fat was rapidly taken and then immersed in a <NUM>% neutral buffered formaldehyde solution for fixation. After washing with water and dehydrating, the cells were treated with a paraffin solution to make a paraffin block. Each paraffin block was cut to a thickness of <NUM>-<NUM>, stained with hematoxylin and eosin, and then observed with an optical microscope. As the results thereof, as shown in <FIG>, mostly intact adipocytes were observed in the groups administered with the formulations of the present invention, whereas the findings of inflammatory reaction were observed in the groups administered with the formulations of Comparative Examples <NUM> and <NUM>.

After the abdomen and the groin of the sacrificed rats were dissected, each subcutaneous fat was rapidly taken and then the fat tissues in the groin were fixed with a <NUM>% formaldehyde solution for <NUM> hours. After dehydrating with a <NUM>% sucrose solution for <NUM> hours, the OCT-embedding cryostats were prepared using cryotome (FSE Cryostats, Thermo Scientific). After slides were prepared in a thickness of <NUM>, adipose tissues were stained with an Oil Red O (Lipid Stain) staining kit (product number: ab150678, Abcam, USA) and the levels of lipolysis were observed with an optical microscope. As the results thereof, as shown in <FIG>, it can be confirmed that the groups administered with the formulations of the present invention exhibited uniform fat distribution, while the groups administered with the formulations of Comparative Examples <NUM> and <NUM> showed aggregated fats, i.e., non-uniform fat distribution. Therefore, it can be confirmed that the formulations of the present invention have a uniform lipolytic activity at the administration site, while the formulations of Comparative Examples <NUM> and <NUM> cannot achieve uniform lipolysis.

Claim 1:
A composition comprising <NUM>,<NUM>-dimyristoyl-sn-glycero-<NUM>-phosphocholine or pharmaceutically acceptable salt thereof for use in ameliorating, inhibiting or treating obesity, wherein the composition is in the form of a pharmaceutical composition for topical administration or in the form of a cosmetic composition for applying to a skin, subcutaneous tissues, muscular tissues, or abdominal tissues.