WO2023059099A1 - Composition pharmaceutique pour la prévention, l'atténuation, le soulagement ou le traitement de la fibrose pulmonaire, comprenant de l'ézétimibe en tant que principe actif - Google Patents

Composition pharmaceutique pour la prévention, l'atténuation, le soulagement ou le traitement de la fibrose pulmonaire, comprenant de l'ézétimibe en tant que principe actif Download PDF

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WO2023059099A1
WO2023059099A1 PCT/KR2022/015046 KR2022015046W WO2023059099A1 WO 2023059099 A1 WO2023059099 A1 WO 2023059099A1 KR 2022015046 W KR2022015046 W KR 2022015046W WO 2023059099 A1 WO2023059099 A1 WO 2023059099A1
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pulmonary fibrosis
ezetimibe
bleomycin
preventing
alleviating
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English (en)
Korean (ko)
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김송이
이찬호
배수한
김영삼
이진구
한지수
임범진
곽세현
신주혜
이유설
박정수
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연세대학교 산학협력단
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/397—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having four-membered rings, e.g. azetidine
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/314—Foods, ingredients or supplements having a functional effect on health having an effect on lung or respiratory system
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/30—Other Organic compounds

Definitions

  • composition for preventing, improving, alleviating or treating pulmonary fibrosis comprising ezetimibe or a salt thereof as an active ingredient, and uses thereof.
  • Pulmonary fibrosis is a disease in which the lungs do not function normally because lung tissue is damaged and scarred to become thick and hard. As pulmonary fibrosis worsens, breathing becomes progressively shorter. In many cases, the exact cause of pulmonary fibrosis cannot be identified. This fibrosis is called idiopathic pulmonary fibrosis (IPF).
  • IPF idiopathic pulmonary fibrosis
  • Pulmonary fibrosis includes idiopathic pulmonary fibrosis; pulmonary fibrosis due to autoimmune diseases such as rheumatoid arthritis, viral infections, diseases such as gastroesophageal reflux disease (GERD); familial pulmonary fibrosis (familial PF); There are various types of pulmonary fibrosis caused by harmful substances such as asbestos, silica, dust, radiation treatment, and smoking. Treatment strategies for pulmonary fibrosis are diverse and individual depending on the condition and symptoms of the patient, but a cure that can cure pulmonary fibrosis has not yet been found.
  • Ezetimibe is a lipid-lowering agent that inhibits cholesterol absorption and is used as a treatment for hyperlipidemia and hypercholesterolemia.
  • Easytrol ® as a single ezetimibe product
  • Vitorin ® ezetimibe + simvastatin
  • Rosuzet ® ezetimibe + rosuvastatin
  • Atojet as combination products of ezetimibe and other hyperlipidemia drugs.
  • ® (ezetimibe + atorvastatin) (Patent Document 1).
  • Patent Document 1 US 2014-0287042 A1
  • compositions for preventing, improving, alleviating or treating pulmonary fibrosis comprising ezetimibe or a pharmaceutically acceptable salt thereof as an active ingredient.
  • a health functional food composition for preventing, improving, or alleviating pulmonary fibrosis comprising ezetimibe or a food chemically acceptable salt thereof as an active ingredient.
  • a method for preventing, ameliorating, alleviating or treating pulmonary fibrosis comprising administering an effective amount of ezetimibe or a pharmaceutically acceptable salt thereof to a subject in need thereof.
  • ezetimibe or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for preventing, ameliorating, alleviating or treating pulmonary fibrosis.
  • ezetimibe or a pharmaceutically acceptable salt thereof for preventing, improving, alleviating or treating pulmonary fibrosis.
  • One aspect provides a pharmaceutical composition for preventing, improving, alleviating or treating pulmonary fibrosis, comprising Ezetimibe or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the “Ezetimibe” has a structural formula represented by Formula 1 below:
  • the ezetimibe was approved by the US Food and Drug Administration (FDA) in 2002, and its substance patent expired in 2016.
  • Ezetimibe can reduce pulmonary fibrosis-related factors induced by TGF- ⁇ , specifically TGF- ⁇ 1, in lung fibroblasts.
  • Ezetimibe can inhibit the differentiation of lung fibroblasts into myofibroblasts or reduce collagen expression.
  • the ezetimibe can inhibit the differentiation of lung fibroblasts into myofibroblasts or reduce the expression of collagen protein and/or mRNA in lung fibroblasts.
  • the collagen may be COL1A1 (collagen, type I, alpha 1).
  • the ezetimibe may be in any pharmaceutically acceptable form.
  • pharmaceutically acceptable means an amount sufficient to exhibit a therapeutic effect and not causing side effects, such as the type of disease, the sensitivity of the patient to the drug, the route of administration, the method of administration, the number of administrations, the duration of treatment, etc. It can be readily determined by a person skilled in the art according to factors well known in the medical arts.
  • the ezetimibe may be in the form of a pharmaceutically acceptable salt thereof.
  • These salts include acid addition salts used in the pharmaceutical field, for example salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid or nitric acid; and acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, citric acid, maleic acid, malonic acid, methanesulfonic acid, tartaric acid, malic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, oxalic acid, salts derived from organic acids such as trifluoroacetic acid or glucuronic acid.
  • inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid or n
  • the salt may be a base addition salt such as ammonium, dimethylamine, monomethylamine, monoethylamine, or diethylamine.
  • the salt includes a common metal salt form, for example, a salt derived from a metal such as lithium, sodium, potassium, magnesium, or calcium.
  • the acid addition salt, base addition salt or metal salt may be prepared according to a conventional method.
  • Pharmaceutically acceptable salts and general methodologies for their preparation are well known in the art. See, for example, P. Stahl, et al. Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2nd Revised Edition (Wiley-VCH, 2011)]; [S.M. Berge, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, Vol. 66, no. 1, January 1977].
  • Ezetimibe is included as an active ingredient in the composition.
  • the term “included as an active ingredient” means that ezetimibe is added to the extent that the above-mentioned effects can be exhibited, and various ingredients are added as subcomponents for drug delivery and stabilization, etc. to be formulated in various forms. It means to include being.
  • the ezetimibe may be included in the composition in a pharmaceutically effective amount.
  • an effective amount refers to that amount or dose of ezetimibe that, when administered to a patient in single or multiple doses, provides a desired effect in a patient under diagnosis or treatment.
  • the effective amount can be readily determined by the attending diagnostician as a person skilled in the art by using known techniques or by observing results obtained under similar circumstances.
  • the mammalian species When determining an effective amount for a patient, the mammalian species; his size, age and general health; the specific disease or disorder involved; degree or severity of involvement of the disease or disorder; individual patient response; the specific compound being administered; administration mode; bioavailability characteristics of the administered agent; the selected dosing regimen; use of concomitant medication; A number of factors are taken into account by the attending physician diagnostician, including but not limited to, and other relevant circumstances.
  • the ezetimibe daily dose is 0.001 to 20 mg, 0.001 to 15 mg, 0.001 to 10 mg, 0.01 to 20 mg, 0.01 to 15 mg, 0.01 to 10 mg, 0.1 to 20 mg, 0.1 to 20 mg It may be included in a dose of 15 mg, or 0.1 to 10 mg, but is not limited thereto.
  • the dose of ezetimibe can be appropriately adjusted according to the individual.
  • pulmonary fibrosis refers to a phenomenon in which a large amount of extracellular matrix including collagen is accumulated in the interstitium of the lung and all diseases in which such a phenomenon appears.
  • the cause of the pulmonary fibrosis is not limited.
  • the pulmonary fibrosis may include pulmonary fibrosis caused by idiopathic interstitial pneumonia of unknown cause (eg, idiopathic pulmonary fibrosis, pulmonary fibrosis caused by idiopathic nonspecific interstitial pneumonia, etc.); pulmonary fibrosis caused by autoimmune diseases or connective tissue diseases such as rheumatoid arthritis, lupus, systemic sclerosis, myositis, Sjogren's syndrome; Pulmonary fibrosis due to diseases such as infectious diseases (eg, coronavirus, pneumocystis pneumonia, etc.), gastroesophageal reflux disease (GERD); familial pulmonary fibrosis (familial PF); pulmonary fibrosis caused by harmful substances such as asbestos, silica, beryllium, dust, and smoking; pulmonary fibrosis due to radiation therapy or radiation exposure; Pulmonary fibrosis caused by drugs such as anticancer drugs (eg Bleomycin), antibiotics, anticancer
  • the pulmonary fibrosis may be idiopathic pulmonary fibrosis.
  • the pulmonary fibrosis may be pulmonary fibrosis caused by administration of bleomycin or sequelae after COVID-19 infection.
  • bleomycin is an anticancer drug used for squamous cell carcinoma of the head and neck, lung, skin, etc., and has been reported to exhibit side effects such as pulmonary fibrosis.
  • the pulmonary fibrosis may include pulmonary fibrosis caused by the use of other drugs such as bleomycin.
  • coronavirus disease is an infectious disease caused by a coronavirus.
  • the coronavirus infection may be COVID-19, but is not limited thereto.
  • coronavirus infection-19 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2: SARS-CoV-2. Therefore, the pulmonary fibrosis may be pulmonary fibrosis caused by COVID-19.
  • the pulmonary fibrosis may be idiopathic pulmonary fibrosis.
  • the pulmonary fibrosis may be pulmonary fibrosis caused by administration of bleomycin or sequelae after COVID-19 infection.
  • bleomycin is an anticancer drug used for squamous cell carcinoma of the head and neck, lung, skin, etc., and has been reported to exhibit side effects such as pulmonary fibrosis.
  • the pulmonary fibrosis may include pulmonary fibrosis caused by the use of other drugs such as bleomycin.
  • prevention refers to any action that inhibits or delays the onset of pulmonary fibrosis by administration of the composition.
  • improvement refers to any process in which symptoms of pulmonary fibrosis are improved by administration of the composition.
  • Alleviation means that the symptoms of pulmonary fibrosis are alleviated, the progression of pulmonary fibrosis is delayed, or the cause(s) of pulmonary fibrosis itself is alleviated or eliminated by administration of the composition. Alleviation within this specification may also include mitigating the progression of pulmonary fibrosis.
  • treatment refers to any action that improves or benefits the symptoms of pulmonary fibrosis by administration of the composition.
  • the pharmaceutical composition may further include a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments and flavors for oral administration, and buffers, preservatives, and painless agents for injections.
  • a topical agent, a solubilizing agent, an isotonic agent, and a stabilizer may be mixed and used, and in the case of topical administration, a base, an excipient, a lubricant, and a preservative may be used.
  • Formulations of the pharmaceutical composition may be variously prepared by mixing with a pharmaceutically acceptable carrier as described above.
  • a pharmaceutically acceptable carrier for example, for oral administration, it can be prepared in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and in the case of injections, it can be prepared in the form of unit dose ampoules or multiple doses.
  • it can be formulated into solutions, suspensions, tablets, pills, capsules, and sustained-release preparations.
  • examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used.
  • fillers, anti-coagulants, lubricants, wetting agents, flavoring agents, emulsifiers and preservatives may be further included.
  • the pharmaceutical composition may further include one or more other agents for treating pulmonary fibrosis.
  • the other agent may be a therapeutic agent for pulmonary fibrosis, but is not limited thereto.
  • the therapeutic agent for pulmonary fibrosis known substances may be used.
  • the therapeutic agent for pulmonary fibrosis may be pirfenidone, nintedanib, and the like.
  • Another aspect provides a health functional food composition for preventing, improving, or alleviating pulmonary fibrosis, comprising Ezetimibe or a food chemically acceptable salt thereof as an active ingredient.
  • the health functional food composition may be formulated into a conventional health functional food formulation known in the art.
  • the health functional food composition may use ezetimibe alone or together with other foods or food ingredients, and may be appropriately used according to conventional methods.
  • the mixing amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).
  • beverage compositions may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages.
  • the natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrins and cyclodextrins; It is a sugar alcohol, such as xylitol, sorbitol, and erythritol.
  • sweetener natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used.
  • the food composition is also used in nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonated beverages carbonation agent, or a combination thereof.
  • the food composition may also contain natural fruit juice, fruit juice beverages, fruit flesh for preparing vegetable beverages, or a combination thereof.
  • Another aspect provides a method for preventing, ameliorating, alleviating or treating pulmonary fibrosis, comprising administering an effective amount of ezetimibe or a pharmaceutically acceptable salt thereof to a subject in need thereof.
  • the term "individual” means a subject in need of treatment for a disease, and more specifically, mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle.
  • the "subject in need thereof” refers to an individual in need of prevention, improvement or treatment of pulmonary fibrosis.
  • administration means the introduction of a substance into a patient by any suitable method.
  • the route of administration may be any general route capable of reaching a target in vivo in a patient.
  • the administration may be, for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, or intrarectal administration, but is not limited thereto.
  • the administration may be oral administration.
  • the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, medical condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can determine these factors Dosage can be appropriately adjusted in consideration of these factors.
  • the dosage ranges from 0.001 mg to 1,000 mg, 0.001 mg to 500 mg, 0.001 mg to 100 mg, 0.001 mg to 50 mg, 0.001 mg to 20 mg, 0.001 mg to 10 mg, ezetimibe per day per subject. It may be 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 20 mg, or 0.1 mg to 10 mg, but is not limited thereto.
  • the number of administrations can be once a day or two or more times within the range of clinically acceptable side effects, and administration can be performed at one or two or more sites, daily or at intervals of 2 to 5 days.
  • the number of administration days may be administered from 1 day to 30 days per treatment. If necessary, the same treatment can be repeated after a titration period.
  • the same dosage per kg as for humans is used, or the above dosage is converted by the volume ratio (eg, average value) of the organ (heart, etc.) between the target animal and the human.
  • a single dose can be administered.
  • an effective amount of ezetimibe or a pharmaceutically acceptable salt thereof may be administered simultaneously, separately, or sequentially with an effective amount of one or more other active ingredients.
  • the one or more other active ingredients may be one or more other agents for treating pulmonary fibrosis, but are not limited thereto.
  • Another aspect provides the use of ezetimibe or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for preventing, ameliorating, alleviating or treating pulmonary fibrosis.
  • Another aspect provides the use of ezetimibe or a pharmaceutically acceptable salt thereof for preventing, ameliorating, alleviating or treating pulmonary fibrosis.
  • Redundant content is omitted in consideration of the complexity of the present specification, and terms not defined otherwise in the present specification have meanings commonly used in the technical field to which the present invention belongs.
  • composition according to one aspect may have an effect of preventing, improving, alleviating or treating pulmonary fibrosis by including ezetimibe or a pharmaceutically acceptable salt thereof. Therefore, ezetimibe can be used as a pharmaceutical composition for preventing, improving, alleviating or treating pulmonary fibrosis.
  • 1 is a result of treating mouse lung fibroblasts with ezetimibe at the indicated concentration for 24 hours in a cell culture medium of 15% FBS, and performing an MTT assay.
  • Figure 2 shows the results of MTT assay after treatment with ezetimibe at the indicated concentration for 24 hours in mouse lung fibroblasts in FBS 0.1% cell culture medium conditions.
  • Figure 3 shows the results of treating human lung fibroblasts with ezetimibe at the indicated concentrations for 24 hours in a cell culture medium of 15% FBS, and performing the MTT assay.
  • Figure 4 is a result of comparing the TGF ⁇ 1 untreated state and the TGF ⁇ 1 treated state, and showing the Col1A1 protein expression level according to the treatment concentration of ezetimibe in the TGF ⁇ 1 treated state at the same time.
  • FIG. 5 is a result showing the mRNA expression level of Col1A1 and Acta2 for each treatment concentration of ezetimibe.
  • FIG. 6 is a result showing the level of Col1A1 protein expression according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M.
  • FIG. 7 is a result showing the mRNA expression levels of Col1A1 and Acta2 according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M.
  • FIG. 10 is a result showing the level of Col1A1 protein expression according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M.
  • FIG. 11 shows the mRNA expression levels of Col1A1 and Acta2 according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M.
  • FIG. 12 is a schematic view showing the administration method of each group in an experiment for confirming the pulmonary fibrosis alleviating effect of ezetimibe in a bleomycin-induced mouse pulmonary fibrosis model.
  • 13 is a graph showing the change in survival rate of mice according to the number of days after administration of bleomycin.
  • 15 is a diagram confirming the results of comparing COL1A1 and fibronectin, which are fibrosis markers of each group, using Western blot and quantifying the result of Western blot through Image J.
  • 16 is a diagram showing the results of confirming the expression of fibrosis markers Col1a1, Col1a2, Acta2, Col3a1 and Eda-fn mRNA through RT-qPCR.
  • 18 is a diagram showing the results of quantification of collagen-stained areas in slides of lung pathological tissues for each mouse using the Aperio Imagescope program.
  • mice In the case of mouse lung tissue, adult mice between 7 and 10 weeks of age were euthanized, and both lungs were aseptically collected.
  • lung tissue of about 2 cm 3 obtained in a general surgical procedure was aseptically removed and used only for patients who agreed to the lung tissue collection study that passed the Institutional Review Board (IRB).
  • IRS Institutional Review Board
  • the culture medium was replaced, and 2 weeks after separation, subculture was performed, and the cell culture medium was changed to MEM with 15% FBS and 1X P/S (penicillin/streptomycin) added.
  • the tissue was removed using a 100 ⁇ m cell filter, and only the isolated lung fibroblasts were cultured. Thereafter, the cells were subcultured at intervals of about 5 days to increase the purity of the cells and increase the number of cells. All experiments were performed while changing to normoxic conditions in the third passage.
  • TGF ⁇ 1 a solution containing 0.1% (w/v) BSA (Bovine Serum Albumin) in a 20 mM citrate solution (pH 3.0) containing 100 mM NaCl was used as a vehicle, and ezetimibe was DMSO ( Dimethyl Sulfoxide) was used as a vehicle. All drugs and their solvents were applied to the cells at 1:1000 (v/v).
  • BSA Bovine Serum Albumin
  • DMSO Dimethyl Sulfoxide
  • Bleomycin is an anticancer drug used for squamous cell carcinoma of the head and neck, lung, skin, etc., and has been reported to exhibit side effects such as pulmonary fibrosis. Based on these characteristics, bleomycin is the most representative drug used in animal models of pulmonary fibrosis. Therefore, pulmonary fibrosis was induced using bleomycin. Experiments were conducted in the following order using C57BL/6J mice between 7 and 10 weeks of age.
  • mice were given a 15-minute fasting period, and anesthesia was performed through respiratory anesthesia.
  • control group control, normal saline
  • 50 ul of aseptically treated physiological saline was aspirated through the pharynx to the mouse, and for the bleomycin-administered group and bleomycin and ezetimibe-administered group, 50 ul of bleomycin 2U/kg was orally administered to the mouse. was aspirated
  • the group administered with bleomycin and ezetimibe received 2 mg/kg of ezetimibe diluted in 0.5% sodium carboxymethyl cellulose prepared in distilled water, and bleomycin and ezetimibe.
  • the group administered with Nintedanib received 40 mg/kg of Nintedanib diluted in 0.5% sodium carboxymethyl cellulose, and the control and bleomycin groups received oral administration of 0.5% sodium carboxymethyl cellulose five times a week, daily.
  • the primary antibody was reacted at 4°C for 18 to 24 hours, washed with TTBS (Tween-Tris buffered saline), and the secondary antibody was reacted at room temperature for more than 1 hour.
  • TTBS Tetween-Tris buffered saline
  • Trizol 1000 ul of Trizol and 200 ul of chloroform were added to the tissues or cells, mixed well, and centrifuged at 13,000 rpm for 15 minutes using a centrifuge at 4 °C.
  • RNA in the solution was quantified using a trace spectrophotometer.
  • cDNA complementary DNA
  • This experiment is an experiment to confirm cytotoxicity by reflecting the number of viable cells by measuring ATP released while lysing cells.
  • the concentration of ezetimibe that causes toxicity to the cells was confirmed using the MTT assay.
  • experiments were conducted in FBS 0.1% culture medium and FBS 15% culture medium, respectively.
  • 1 is a result of treating mouse lung fibroblasts with ezetimibe at the indicated concentration for 24 hours in a cell culture medium of 15% FBS, and performing an MTT assay.
  • Figure 2 shows the results of MTT assay after treatment with ezetimibe at the indicated concentration for 24 hours in mouse lung fibroblasts in FBS 0.1% cell culture medium condition.
  • Figure 3 shows the results of treating human lung fibroblasts with ezetimibe at the indicated concentrations for 24 hours in a cell culture medium of 15% FBS, and performing the MTT assay.
  • ezetimibe did not cause cytotoxicity in human and mouse lung fibroblasts at a concentration of less than 20 ⁇ M. Based on this, in subsequent experiments, 20 ⁇ M of ezetimibe was used at the maximum concentration.
  • Collagen 1a1 (Col1A1), one of the final products of fibrosis produced by treating fibroblasts with TGF ⁇ 1 to differentiate into myofibroblasts, was identified as a major detection target.
  • Figure 4 is a result of comparing the TGF ⁇ 1 untreated state and the TGF ⁇ 1 treated state, and showing the Col1A1 protein expression level according to the treatment concentration of ezetimibe in the TGF ⁇ 1 treated state at the same time.
  • FIG. 5 shows the mRNA expression levels of Col1A1 and Acta2 for each treatment concentration of ezetimibe under the same experimental conditions as in FIG. 4 .
  • FIG. 6 is a result showing the level of Col1A1 protein expression according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M.
  • FIG. 7 is a result showing the mRNA expression levels of Col1A1 and Acta2 according to the treatment time of ezetimibe when the concentration of ezetimibe was fixed at 20 ⁇ M under the same experimental conditions as in FIG. 6 .
  • Collagen 1a1 (Col1A1), one of the final products of fibrosis produced by treating fibroblasts with TGF ⁇ 1 to differentiate into myofibroblasts, was identified as a major detection target.
  • FIG. 9 shows the mRNA expression levels of Col1A1 and Acta2 for each treatment concentration of ezetimibe under the same experimental conditions as in FIG. 8 .
  • FIG. 10 is a result showing the level of Col1A1 protein expression according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M.
  • FIG. 11 is a result showing the mRNA expression levels of Col1A1 and Acta2 according to ezetimibe treatment time when the concentration of ezetimibe was fixed at 20 ⁇ M under the same experimental conditions as in FIG. 10.
  • the TGF ⁇ 1-treated group also increased COL1A1 protein and mRNA expression by TGF ⁇ stimulation, but such an increase in COL1A1 protein and mRNA expression was effectively expressed by ezetimibe treatment It was confirmed that this decreased.
  • ezetimibe has a therapeutic effect on pulmonary fibrosis in human and mouse pulmonary fibrosis cell models by TGF ⁇ 1 stimulation of pulmonary fibroblasts.
  • mice In order to confirm the effect of ezetimibe on alleviating pulmonary fibrosis in a bleomycin-induced mouse pulmonary fibrosis model, the survival rate of mice was confirmed.
  • FIG. 12 is a schematic view showing the administration method of each group in an experiment for confirming the pulmonary fibrosis alleviating effect of ezetimibe in a bleomycin-induced mouse pulmonary fibrosis model.
  • 13 is a graph showing the change in survival rate of mice according to the number of days after administration of bleomycin.
  • mice administered with ezetimibe exhibited an excellent survival effect compared to mice administered with bleomycin, which was found to be similar to that of nintedanib, an existing idiopathic pulmonary fibrosis drug.
  • 15 is a diagram confirming the results of comparing COL1A1 and fibronectin, which are fibrosis markers of each group, using Western blot and quantifying the result of Western blot through Image J.
  • 16 is a diagram showing the results of confirming the expression of fibrosis markers Col1a1, Col1a2, Acta2, Col3a1 and Eda-fn mRNA through RT-qPCR.
  • 18 is a diagram showing the results of quantification of collagen-stained areas in slides of lung pathological tissues for each mouse using the Aperio Imagescope program.
  • ezetimibe showed efficacy in inhibiting fibrosis in mice administered with bleomycin, especially when compared to mice using standard dose of nintedanib, a drug already marketed and used internationally for patients with idiopathic pulmonary fibrosis. However, it was confirmed that similar or superior effects appeared. Accordingly, it was confirmed that ezetimibe can alleviate and treat pulmonary fibrosis.

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  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Mycology (AREA)
  • Nutrition Science (AREA)
  • Polymers & Plastics (AREA)
  • Food Science & Technology (AREA)
  • Epidemiology (AREA)
  • Pulmonology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne : une composition pharmaceutique pour la prévention, l'atténuation, le soulagement ou le traitement de la fibrose pulmonaire, la composition comprenant de l'ézétimibe ou un sel de celui-ci en tant que principe actif ; une composition alimentaire fonctionnelle pour la santé ; une méthode pour la prévention, l'atténuation, le soulagement ou le traitement de la fibrose pulmonaire l'utilisant ; et une utilisation de celles-ci. La composition selon un aspect de la présente invention contient de l'ézétimibe et peut ainsi avoir pour effet de prévenir, d'atténuer, de soulager ou de traiter la fibrose pulmonaire.
PCT/KR2022/015046 2021-10-08 2022-10-06 Composition pharmaceutique pour la prévention, l'atténuation, le soulagement ou le traitement de la fibrose pulmonaire, comprenant de l'ézétimibe en tant que principe actif WO2023059099A1 (fr)

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JP2017203013A (ja) * 2016-05-13 2017-11-16 エルメッド エーザイ株式会社 エゼチミブ含有医薬組成物及びその製造方法、エゼチミブ含有医薬組成物の硬度低下抑制剤及び硬度低下抑制方法、並びにエゼチミブ含有医薬組成物の吸湿抑制剤及び吸湿抑制方法
KR101845862B1 (ko) * 2015-11-11 2018-04-06 (주)나디안바이오 특발성 폐섬유증의 치료 또는 예방을 위한 약학적 조성물
JP2018184381A (ja) * 2017-04-27 2018-11-22 高田製薬株式会社 エゼチミブ含有錠剤およびその製法
KR102001437B1 (ko) * 2019-02-25 2019-07-19 연세대학교 산학협력단 섬유증의 예방 또는 치료용 약학 조성물
KR102289381B1 (ko) * 2020-03-17 2021-08-17 주식회사 대웅테라퓨틱스 당뇨치료제 및 고지혈증 치료제를 포함하는 약학적 조성물

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9763885B2 (en) 2012-05-01 2017-09-19 Althera Laboratories Ltd. Oral tablet formulation consisting of fixed combination of rosuvastatin and ezetimibe for treatment of hyperlipidemia and cardiovascular diseases

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101845862B1 (ko) * 2015-11-11 2018-04-06 (주)나디안바이오 특발성 폐섬유증의 치료 또는 예방을 위한 약학적 조성물
JP2017203013A (ja) * 2016-05-13 2017-11-16 エルメッド エーザイ株式会社 エゼチミブ含有医薬組成物及びその製造方法、エゼチミブ含有医薬組成物の硬度低下抑制剤及び硬度低下抑制方法、並びにエゼチミブ含有医薬組成物の吸湿抑制剤及び吸湿抑制方法
JP2018184381A (ja) * 2017-04-27 2018-11-22 高田製薬株式会社 エゼチミブ含有錠剤およびその製法
KR102001437B1 (ko) * 2019-02-25 2019-07-19 연세대학교 산학협력단 섬유증의 예방 또는 치료용 약학 조성물
KR102289381B1 (ko) * 2020-03-17 2021-08-17 주식회사 대웅테라퓨틱스 당뇨치료제 및 고지혈증 치료제를 포함하는 약학적 조성물

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