WO2023014049A1 - Novel pyrrolopyrimidine derivative, method of preparing same, and pharmaceutical composition comprising same as active ingredient for preventing or treating metabolic liver disease - Google Patents

Novel pyrrolopyrimidine derivative, method of preparing same, and pharmaceutical composition comprising same as active ingredient for preventing or treating metabolic liver disease Download PDF

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WO2023014049A1
WO2023014049A1 PCT/KR2022/011400 KR2022011400W WO2023014049A1 WO 2023014049 A1 WO2023014049 A1 WO 2023014049A1 KR 2022011400 W KR2022011400 W KR 2022011400W WO 2023014049 A1 WO2023014049 A1 WO 2023014049A1
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formula
compound represented
substituted
liver disease
chemical formula
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PCT/KR2022/011400
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French (fr)
Korean (ko)
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배명애
안진희
양정윤
신대섭
황규석
김성순
이병회
안세환
김동건
허예림
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한국화학연구원
광주과학기술원
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Publication of WO2023014049A1 publication Critical patent/WO2023014049A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/116—Heterocyclic compounds
    • A23K20/137—Heterocyclic compounds containing two hetero atoms, of which at least one is nitrogen
    • 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
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • 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
    • 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

  • the present invention relates to a novel pyrrolopyrimidine derivative compound, a preparation method thereof, and a pharmaceutical composition for preventing or treating metabolic liver disease comprising the same as an active ingredient.
  • Fatty liver refers to a state in which fat is accumulated in liver cells, and medically refers to a pathological condition in which fat exceeds 5% of the total weight of the liver. It is recognized as the second most serious disease after cancer. 30% of the population of major countries, including developed countries, show fatty liver symptoms, and 20% of them progress to cirrhosis, and about half of cirrhosis patients die of liver disease within 10 years after diagnosis.
  • Fatty liver can be largely divided into alcoholic fatty liver disease caused by excessive drinking and nonalcoholic fatty liver disease (NAFLD) caused by obesity, diabetes, hyperlipidemia, and drugs.
  • NAFLD nonalcoholic fatty liver disease
  • Non-alcoholic fatty liver disease refers to simple fatty liver without inflammatory reactions in patients who have not consumed excessive alcohol, and a wide range of diseases that are developed by it and include inflammatory reactions of hepatocytes, liver fibrosis, and liver cirrhosis.
  • Nonalcoholic fatty liver disease is divided into primary and secondary depending on the cause. The primary is caused by hyperlipidemia, diabetes, or obesity, which are characteristic of metabolic syndrome, and the secondary is caused by nutritional causes (rapid weight loss, starvation, intestinal bypass), various drugs, It is known to be caused by toxic substances (poisonous mushrooms, bacterial toxins), metabolic causes, and other factors.
  • NAFLD non-alcoholic fatty liver disease
  • Orlistat which is used as an oral obesity treatment, has been reported to induce histological improvement of the liver in patients with steatohepatitis.
  • Thiazolidinedione (TZD) drugs which are agonists of PPAR (peroxisome proliferator-activated receptor), inhibit fat accumulation in the liver and muscle and have a direct effect on the liver in animal models of non-alcoholic fatty liver disease. It has been reported to exhibit anti-fibrotic activity.
  • PPAR peroxisome proliferator-activated receptor
  • An object to be solved by the present invention is to provide a novel pyrrolopyrimidine derivative compound.
  • the present invention provides a method for preparing the novel pyrrolopyrimidine derivative compound.
  • the present invention is to provide a composition for preventing, treating or improving metabolic liver disease comprising the novel pyrrolopyrimidine derivative compound as an active ingredient.
  • the present invention provides a method for treating metabolic liver disease, comprising administering the novel pyrrolopyrimidine derivative compound in a therapeutically effective amount to a subject.
  • the present invention provides a pyrrolopyrimidine derivative compound represented by Formula 1 below, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
  • R 1 is N-acetylalanine, or substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
  • X is sulfinyl or sulfanyl.
  • the present invention comprises the steps of preparing a compound represented by Formula 1 from a compound represented by Formula 5; A method for preparing a pyrrolopyrimidine derivative compound represented by Formula 1 including a is provided.
  • R 1 is N-acetylalanine, or substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
  • X is sulfinyl or sulfanyl.
  • the present invention provides a pharmaceutical composition for preventing or treating metabolic liver disease comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a food composition for preventing or improving metabolic liver disease comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides an animal feed composition for preventing or improving metabolic liver disease, comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the present invention provides a method for treating metabolic liver disease, comprising administering a therapeutically effective amount of the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof to a subject.
  • novel pyrrolopyrimidine derivative compound according to the present invention can inhibit the accumulation of fat, so a pharmaceutical composition, food composition or feed that can prevent, treat or improve metabolic liver disease, especially fatty liver, by including it as an active ingredient composition may be provided.
  • FIG. 2 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 1 using Gen5 software.
  • FIG. 3 is a fluorescence image showing changes in fat droplets in liver organoids after treatment with the compounds prepared in Examples 1 and 2.
  • FIG. 4 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 2 using Gen5 software.
  • Example 5 is a result of inhibition of fatty liver formation in a tamoxifen-induced fatty liver model by the compound prepared in Example 1.
  • Figure 6 is the compound prepared in Example 2 This is the result of inhibition of fatty liver formation in the tamoxifen-induced fatty liver model.
  • a pyrrolopyrimidine derivative compound represented by Formula 1 below, an optical stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
  • R 1 is N-acetylalanine, or substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
  • X is sulfinyl or sulfanyl.
  • the “halogen” may be fluoro, chloro, bromo or iodine, preferably fluoro.
  • the R 1 may be N-acetylalanine or phenyl substituted with halogen.
  • the compound represented by Formula 1 may be a compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that it is any one selected from the group of compounds below:
  • the compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful as the salt.
  • Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes.
  • non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids, etc.
  • organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like.
  • pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, i.
  • the acid addition salt according to the present invention can be prepared by a conventional method, for example, dissolving the pyrrolo pyrimidine derivative compound of Formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile, etc., and adding an organic acid or It can be prepared by filtering and drying the precipitate produced by adding an inorganic acid, or by distilling the solvent and excess acid under reduced pressure, drying it, and crystallizing it in an organic solvent.
  • an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile, etc.
  • a pharmaceutically acceptable metal salt may be prepared using a base.
  • An alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt.
  • the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).
  • the present invention includes not only the pyrrolopyrimidine derivative compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like prepared therefrom.
  • R 1 is N-acetylalanine, or substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
  • X is sulfinyl or sulfanyl.
  • Step 1 preparing a compound represented by Chemical Formula 3 by reacting a compound represented by Chemical Formula 5 with a compound represented by Chemical Formula 4 (Step 1); and preparing a compound represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone (Step 2); It may contain.
  • R 1a is substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen.
  • Step 1 preparing a compound represented by Chemical Formula 3 by reacting a compound represented by Chemical Formula 5 with a compound represented by Chemical Formula 4 (Step 1);
  • Step 2 preparing a compound represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone (Step 2);
  • Step 3 preparing a compound represented by Chemical Formula 1-2 by reacting the compound represented by Chemical Formula 1-1 prepared in Step 2 with the compound represented by Chemical Formula 2 (Step 3); It may contain.
  • R 1a is substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
  • R 1b is N-acetylalanine.
  • Step 1 of Scheme 2 or Scheme 3 is a step of preparing a compound represented by Formula 3 by reacting a compound represented by Formula 5 with a compound represented by Formula 4. am.
  • Step 1 can be understood as a step of introducing a -SR 1a substituent into pyrrolopyrimidine, which is the parent nucleus structure of the compound of the present invention, and the reaction time and reaction temperature of step 1 can be determined by referring to the following examples. Any range that can be easily changed by those skilled in the art is included in the scope of the present invention without limitation. In particular, it should be understood that any condition under which the compound represented by Formula 3 to be prepared in step 1 can be prepared is not limited and is included within the scope of the present invention. As just one example, preferably, the reaction time may be carried out for 5 to 20 hours, and the reaction temperature may be carried out at 120 to 140 ° C.
  • a compound represented by Formula 5 and a compound represented by Formula 4 are refluxed together with triethylamine (TEA) or N,N-diisopropylethylamine (DIPEA) in a solvent to obtain Formula 3 compounds can be prepared.
  • the triethylamine (TEA) or N,N-diisopropylethylamine (DIPEA) was used as a base for the nucleophilic aromatic substitution reaction.
  • H 2 O H 2 O, methanol, ethanol, propanol, butanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylformamide, etc.
  • THF tetrahydrofuran
  • dichloromethane dichloromethane
  • toluene acetonitrile
  • dimethylformamide etc.
  • step 2 of Reaction Scheme 2 or Scheme 3 is represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone. This is the step of preparing a compound to be.
  • Step 2 can be understood as a step of introducing a sulfinyl group into pyrrolopyrimidine, which is the parent nuclear structure of the compound of the present invention, and the reaction time and reaction temperature of step 2 can be understood by those skilled in the art with reference to the following examples. Any range that can be easily changed is included in the scope of the present invention without limitation. In particular, it should be understood that any condition under which the compound represented by Formula 1-1 to be prepared in step 2 can be prepared is not limited and is included within the scope of the present invention. As just one example, preferably, the reaction time may be carried out for 5 to 20 hours, and the reaction temperature may be carried out at 0 to 30 °C. As a non-limiting example, after dissolving the compound represented by Chemical Formula 3 in a solvent, the compound represented by Chemical Formula 1-1 may be prepared by mixing and reacting with oxone dissolved in another solvent.
  • H 2 O methanol, ethanol, propanol, butanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylformamide, etc.
  • H 2 O, tetrahydrofuran (THF) or a mixture thereof can be used, and when using a mixture of H 2 O and tetrahydrofuran (THF), the ratio of H 2 O: tetrahydrofuran (THF) is 1: 2 can be used in combination with
  • the present invention can prepare the compound represented by Chemical Formula 1-2 by performing step 3 of Reaction Scheme 3 as well as preparing the compound represented by Chemical Formula 1-1 in the preparation method represented by Reaction Scheme 2 above. there is.
  • Step 3 of Reaction Scheme 3 is a step for preparing a compound represented by Chemical Formula 1-2 by reacting the compound represented by Chemical Formula 1-1 prepared in Step 2 with the compound represented by Chemical Formula 2.
  • the compound represented by Chemical Formula 1-2 can react the compound represented by Chemical Formula 5 in Step 1 with the compound represented by Chemical Formula 2 in Step 3 to prepare the compound represented by Chemical Formula 1-2
  • the reaction is not completely converted, so the yield of the compound represented by Chemical Formula 1-2 is very low. Therefore, in order to increase the yield of the compound represented by Chemical Formula 1-2, the chlorine of the compound represented by Chemical Formula 5 is substituted with a sulfanyl (-S-) group through Step 1, and the compound represented by Chemical Formula 3 is obtained through Step 2.
  • the compound represented by Chemical Formula 1-2 may be prepared through step 3.
  • Step 3 can be understood as a step of introducing a -SR 1b substituent into pyrrolopyrimidine, which is the parent nuclear structure of the compound of the present invention, and the reaction time and reaction temperature of step 3 can be determined by referring to the following examples. Any range that can be easily changed by those skilled in the art is included in the scope of the present invention without limitation. In particular, it should be understood that any condition under which the compound represented by Formula 1-2 to be prepared in step 3 can be prepared is not limited and is included within the scope of the present invention. As just one example, preferably, the reaction time may be carried out for 5 to 20 hours, and the reaction temperature may be carried out at 0 to 30 °C. As a non-limiting example, the compound represented by Chemical Formula 1-2 may be prepared by dissolving the compound represented by Chemical Formula 1-1 in a solvent, adding the compound represented by Chemical Formula 2 and reacting the compound represented by Chemical Formula 1-2.
  • H 2 O methanol, ethanol, propanol, butanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylformamide, etc.
  • THF tetrahydrofuran
  • dichloromethane dichloromethane
  • toluene acetonitrile
  • dimethylformamide etc.
  • aceto Nitrile may be used.
  • the compound obtained as a result of the reaction in the present invention may be isolated through general filtration, washing, drying, and concentration steps used in the field of organic chemistry.
  • compositions for preventing or treating metabolic liver disease comprising a pyrrolopyrimidine derivative represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • R 1 is N-acetylalanine, or substituted or unsubstituted phenyl
  • the substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
  • X is sulfinyl or sulfanyl.
  • the compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt may be understood as an active ingredient for inhibiting fat accumulation.
  • the metabolic liver disease may be alcoholic steatohepatitis or non-alcoholic steatohepatitis, specifically, the non-alcoholic fatty liver disease is simple fatty liver disease, nutritional fatty liver disease, starvation fatty liver disease, obesity fatty liver disease, diabetic fatty liver disease and steatohepatitis. It may be one or more selected from the group consisting of.
  • the pyrrolopyrimidine derivative compound represented by Formula 1, its isomer or pharmaceutically acceptable salt of the present invention can exhibit medically useful effects such as prevention, improvement, and treatment of the disease, This is supported by the present specification, examples and experimental examples, and a pharmaceutical composition containing it as an active ingredient can be provided.
  • the amount of the active ingredient, the pyrrolopyrimidine derivative compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt, according to the mode and method of use of the pharmaceutical composition of the present invention is determined by those skilled in the art. It can be used by properly adjusting.
  • the pharmaceutical composition comprises 0.01 to 50% by weight of the pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, based on the total weight of the total composition. , preferably 0.1 to 25% by weight, more preferably 0.1 to 10% by weight.
  • the pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof may be included alone in the pharmaceutical composition, or other pharmaceutically acceptable carriers, excipients, diluents or It may also be included with sub-components.
  • Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , dextrin, calcium carbonate, propylene glycol, liquid paraffin, and at least one selected from the group consisting of physiological saline, but is not limited thereto, and all conventional carriers, excipients, or diluents may be used.
  • the pharmaceutical composition may include conventional fillers, extenders, binders, disintegrants, anti-agglomerates, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, pectic acid and its salts, protective chlorides, Glycerin, flavors, emulsifiers or preservatives may be further included.
  • the pharmaceutical composition may include one or more other therapeutic agents known to be effective for the treatment or prevention of metabolic liver disease in addition to the active ingredient, and may be used as a combination therapy applied simultaneously or at different times.
  • the method of administration of the pharmaceutical composition can be either oral or parenteral, and for example, it can be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular.
  • the formulation of the composition may vary depending on the method of use, and is formulated using a method well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be.
  • solid preparations for oral administration include tablets (TABLETS), pills, soft or hard capsules (CAPSULES), pills (PILLS), powders (POWDERS) and granules (GRANULES), etc.
  • these preparations include one or more Excipients, for example, may be prepared by mixing starch, calcium carbonate, sucrose or lactose, gelatin, and the like.
  • lubricants such as magnesium stearate and talc may also be used.
  • Liquid formulations for oral use include suspensions, solutions for oral use, emulsions, and syrups.
  • compositions for parenteral administration include creams, lotions, ointments, PLASTERS, LIQUIDS AND SOULTIONS, aerosols, FRUIDEXTRACTS, and elixirs. It may be in the form of (ELIXIR), INFUSIONS, SACHET, PATCH, or INJECTIONS, and may be in the form of an isotonic aqueous solution or suspension preferably in the case of an injectable formulation. .
  • the pharmaceutical composition may further contain adjuvants such as sterilizers, preservatives, stabilizers, hydration agents or emulsification accelerators, salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, and may be conventionally mixed or granulated. It can be formulated according to a coating or coating method, and in addition, it can be formulated using an appropriate method known in the art.
  • adjuvants such as sterilizers, preservatives, stabilizers, hydration agents or emulsification accelerators, salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, and may be conventionally mixed or granulated. It can be formulated according to a coating or coating method, and in addition, it can be formulated using an appropriate method known in the art.
  • the dosage of the pharmaceutical composition may be determined in consideration of the administration method, the age and sex of the user, the severity of the patient, the condition, the absorption of the active ingredient in the body, the inactivity rate, and the drugs used in combination, once or several times. It can be administered in divided doses.
  • an active ingredient of the pharmaceutical composition preferably in an amount of 0.001 to 100 mg/kg body weight, preferably 0.01 to 35 mg/kg body weight, on a daily basis to mammals including humans, once a day or in divided doses orally or It can be administered by the parenteral route.
  • the treatment of metabolic liver disease comprising administering to a subject a therapeutically effective amount of a pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. provides a treatment method.
  • the treatment method may further include a step of identifying a patient in need of prevention or treatment of the metabolic liver disease prior to the administration step.
  • the "therapeutically effective amount” means an amount of an active ingredient for mammals effective for preventing or treating metabolic liver disease, and the therapeutically effective amount is the type of disease, the severity of the disease, the active ingredient and other ingredients contained in the composition It can be adjusted according to various factors, including the type and content of the formulation and the patient's age, weight, general health condition, sex and diet, administration time, administration route and blood clearance of the composition, treatment period, and concurrently used drugs. However, preferably, as described above, 0.001 to 100 mg / kg body weight per day, preferably 0.01 to 35 mg / kg body weight, administered once a day or divided by oral or parenteral route can do.
  • the “subject” may refer to mammals such as humans or non-human primates, mice, dogs, cats, horses, and cows, but is not limited thereto.
  • a food composition for preventing or improving metabolic liver disease comprising the compound represented by Formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
  • the food composition is meant to include all forms of functional food, nutritional supplements, health food, food additives, or feed. , humans or animals, including livestock, are intended as food. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
  • Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
  • General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruits, bottled products, jams, marmalades, etc.), fish, meat and their processed foods (e.g. ham, sausages) Corned beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g.
  • the pyrrolopyrimidine derivative compound represented by Formula 1 of the present invention, its optical isomer or its pharmaceutically acceptable salt It can be prepared by adding
  • nutritional supplements are not limited thereto, but may be prepared by adding the compound of Formula 1 of the present invention to capsules, tablets, pills, etc.
  • health functional foods are not limited thereto, but, for example, the pyrrolopyrimidine derivative compound represented by Formula 1 of the present invention, its optical isomer or its pharmaceutically acceptable salt in the form of tea, juice and drink It can be consumed by liquefying, granulating, encapsulating, and powdering so that it can be manufactured and consumed (health drink).
  • the health food of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol or a carbonating agent, and the like.
  • the health food of the present invention may contain fruit flesh for producing natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
  • An animal feed composition for preventing or improving metabolic liver disease comprising a pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient is provided.
  • the term 'feed' refers to any natural or artificial diet, one-meal meal, etc., or a component of the one-meal meal, suitable for or suitable for animals to eat, ingest, and digest, and to prevent metabolic liver disease according to the present invention.
  • the feed containing the composition for improvement as an active ingredient can be prepared from various types of feed known in the art, and preferably includes concentrated feed, roughage and/or special feed, but is not limited thereto.
  • the term 'feed additive' is a substance added to feed for various purposes, such as supplementation of nutrients and prevention of weight loss, enhancement of digestibility of fiber in feed, improvement of oil quality, prevention of reproductive disorders and improvement of conception rate, and prevention of high-temperature stress in summer.
  • the feed additive of the present invention corresponds to supplementary feed under the Feed Control Act, and includes mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium, and kerotene.
  • vitamins such as vitamins A, D, E, nicotinic acid, and vitamin B complex
  • protective amino acids such as methionine and lysine
  • protected fatty acids such as calcium salts of fatty acids
  • probiotics lactic acid bacteria
  • yeast cultures live bacteria such as mold fermented products
  • Yeast agents and the like may be further included.
  • Concentrated feed includes seed fruits including grains such as wheat, oats, and corn, bran including rice bran, wheat bran, and barley bran as by-products obtained after refining grains, soybeans, fluids, sesame seeds, linseed, and coco palm oil.
  • Fish soluble which is a condensed fresh liquid obtained from fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal soluble
  • meat meal blood meal, feather meal, skim milk powder, cheese from milk, dried whey, which is the balance when casein is produced from skim milk, and animal feed such as yeast, chlorella, and seaweed.
  • animal feed such as yeast, chlorella, and seaweed.
  • Silage which is a stored feed filled and fermented with lactic acid, grass, hay dried by cutting grass, straw of breeding crops, and leaves of leguminous plants, but are not limited thereto.
  • Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivative, diureide isobutane, and supplements for ingredients that tend to be insufficient when only natural feed ingredients are mixed, or formulated feeds to improve the storage life of feeds.
  • feed additives and dietary supplements which are substances added in small amounts, but are not limited thereto.
  • the feed additive for preventing or improving metabolic liver disease according to the present invention is a pyrrolopyrimidine derivative compound represented by Formula 1 of the present invention, an optical isomer thereof, in an appropriate effective concentration range according to various feed manufacturing methods known in the art Or it can be prepared by adding a pharmaceutically acceptable salt thereof.
  • the obtained compound was purified by silica gel column chromatography using ethyl acetate/dichloromethane to obtain the desired pure compound, 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d ] pyrimidine (700 mg / 44.4%).
  • the obtained compound was purified by silica gel column chromatography using ethyl acetate/dichloromethane to obtain the desired pure compound, 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d ] pyrimidine (700 mg / 44.4%).
  • HepG2 cells used DMEM medium, and 10% FBS (Fetal Bovine Serum) and 100 ⁇ g/mL antibiotics were added to the medium. The cells were cultured in a 37°C incubator maintained at 5% CO 2 for 24 hours, and when they reached 70-80% growth every 2-3 days, they were washed with PBS (Phosohate buffer solution), and Trypsin-EDTA (Gibco, USA). ) and subcultured. HepG2 cells were prepared in a 24-well plate at 5 x 10 4 cells/well and stabilized in a 37°C incubator maintained at 5% CO 2 for 24 hours. To induce steatosis, oleic acid and palmitic acid were treated at a ratio of 2:1 to a final concentration of 1 mM.
  • FBS Fetal Bovine Serum
  • Example 1 is an image showing the change in fat droplets in HepG2 human hepatocytes by fluorescence after treatment with the compounds prepared in Examples 1 and 2 according to Experimental Example 1. As the concentration of the compound prepared in Examples 1 and 2 increases, It can be confirmed that the accumulation of fat droplets increased by the fatty acid mixture is reduced.
  • FIG. 2 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 1 using Gen5 software. As a result of fat staining, it was confirmed that Nile red fluorescence intensity was suppressed by 74% at 1 ⁇ M concentration of the compound prepared according to Example 1 and decreased by 70% at 5 ⁇ M concentration of the compound prepared according to Example 2 compared to the steatosis induction group. .
  • Human-derived liver organoids were cultured in a general growth medium.
  • the growth medium was advanced DMEF/F-12 medium containing 1% penicilin-streptomycin, 1% Glutamax, 10 mM HEPES, 1x N2, 1x B27 (w/o vitA), 1% insulin-Transferrin-Selenium, and 50 ng/mL EGF.
  • Liver organoids were cultured for 7 days by changing the growth medium every 2-3 days and adapting them in a humidified incubator at 37°C and 5% CO 2 .
  • hepatic organoids were fragmented by pipetting 10 to 15 times using a 1 mL micropipette, diluted with matrigel so that the number of fragments was 100 to 200, and ultra low binding 96- The wells were cultured for 3 days.
  • liver organoids were treated with a fatty acid mixture solution of oleic acid and plamitic acid at a ratio of 2:1 to a concentration of 1 mM.
  • GM-90383 and GM-90803 of the present invention were prepared to be 10 mM using DMSO, then serially diluted, treated to the final concentration in each well, and cultured for 24 hours.
  • nile-red was mixed with the medium to a final concentration of 10 ⁇ M, and nuclei were simultaneously stained with 5 ⁇ g/mL Hoechst33342 to set the region to be analyzed. After carrying out staining for about 15 minutes, it was washed three times with HM culture medium to remove compounds and staining reagents.
  • Liver organoid photographs were acquired using Gen5 software on Bio-Tek's Lion-Heart instrument.
  • the fluorescence image of Hoechst33342 stained cell nuclei was obtained using a DAPI filter (Excitation 377/50 nm, Emmision 447/60 nm), and the fluorescence image of nile-red stained fat droplets was obtained using an RFP filter (Excitation 531/40 nm, Emmison 593/40 nm).
  • Quantitative analysis of fat droplets accumulated inside the liver organoid was performed using Gen5 software by measuring the nile-red fluorescence intensity within the fluorescence region of Hoechst33342.
  • FIG. 3 is an image showing changes in fat droplets in liver organoids by fluorescence after treatment with the compounds prepared in Examples 1 and 2 according to Experimental Example 2; As the concentration of the compound prepared in Examples 1 and 2 of the present invention increases, it can be confirmed that the accumulation of fat droplets increased by the fatty acid mixture is reduced.
  • FIG. 4 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 3 using Gen5 software.
  • the compound prepared in Example 2 of the present invention reduced the accumulation of fat droplets increased by the fatty acid mixture solution, and in particular, reduced the accumulation of fat droplets most significantly at 5 ⁇ M.
  • the compounds prepared in Example 2 were shown to significantly reduce the accumulation of fat droplets at concentrations of 1 and 5 ⁇ M.
  • Tamoxifen is a non-steroidal anti-estrogen drug that binds to the estrogen receptor, developed by the British ICI company, and has been used most often as an endocrine therapy for female hormone-dependent cancer since the 1980s.
  • zebrafish fry treated with tamoxifen are known to show hepatocyte necrosis and steatosis.
  • the fatty liver inhibitory efficacy of the compounds prepared in Examples 1 and 2 was evaluated using a tamoxifen-induced fatty liver model. want to do
  • the 24-well plate into which the compound prepared in Examples 1 and 2 was put was wrapped with foil to block light, and then incubated in a 28° C. incubator for 24 hours.
  • To remove the compounds in each well wash with egg water 3 times for 5 minutes.
  • fish LipidGreen2 was diluted to 5 uM and exposed to fry for 30 minutes, followed by washing with egg water three times for 5 minutes.
  • Tricaine 5 g/l
  • the fry were transferred to 3% methyl cellulose and photographed.
  • Lionheart FX Automated Microscope (BioTek) and Gen5 (BioTek) software were used.
  • the fluorescence wavelength used was a GFP filter set (Excitation 469 nm, Emmision 525 nm).
  • the liver area and the degree of fat staining of fry on the image were quantified in Gen5 software, and converted to heat map images using ImageJ 1.50i software (National Institutes of Health, USA). Point scattering on the fluorescence microscopy image and the heat map conversion image indicated the liver of the same fry. it means.
  • the graph at the bottom of each image is a graph showing the results of determining the area and fluorescence intensity of the liver (p value is the result of the t-test for each experimental group. *p ⁇ 0.05, **p ⁇ 0.01, ***p ⁇ 0.001).
  • Example 5 is a result of inhibition of fatty liver formation in a tamoxifen-induced fatty liver model by Compound B prepared in Example 1.
  • the fluorescence intensity increased by about 41% compared to the control group.
  • the compound prepared in Example 1 showed a decrease in fluorescence intensity of about 38% in the 1 uM treatment group and about 30% in the 2 uM treatment group, indicating that it can be usefully used for the treatment or alleviation of fatty liver.
  • Example 6 is a result of inhibition of fatty liver formation in a tamoxifen-induced fatty liver model by the compound prepared in Example 2.
  • the fluorescence intensity of the tamoxifen-induced fatty liver model is increased by about 53% compared to the control group.
  • the fluorescence intensity decreased by about 54% in the compound C 1 uM treatment group and by about 43% in the 2 uM treatment group, indicating that it can be usefully used for the treatment or alleviation of fatty liver.

Abstract

The present invention relates to a novel pyrrolopyrimidine derivative compound, a preparation method therefor, and a pharmaceutical composition comprising same as an active ingredient for prevention or treatment of metabolic liver disease. The novel pyrrolopyrimidine derivative compound according to the present invention can inhibit the accumulation of lipids and can serve as an active ingredient for use in preventing, treating, or alleviating metabolic liver diseases.

Description

신규한 피롤로 피리미딘 유도체 화합물, 이의 제조방법, 및 이를 유효성분으로 포함하는 대사성 간질환의 예방 또는 치료용 약학적 조성물Novel pyrrolopyrimidine derivative compound, preparation method thereof, and pharmaceutical composition for preventing or treating metabolic liver disease containing the same as an active ingredient
본 발명은 신규한 피롤로 피리미딘 유도체 화합물, 이의 제조방법, 및 이를 유효성분으로 포함하는 대사성 간질환의 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a novel pyrrolopyrimidine derivative compound, a preparation method thereof, and a pharmaceutical composition for preventing or treating metabolic liver disease comprising the same as an active ingredient.
지방간은 간 세포 속에 지방이 축적된 상태를 의미하며, 의학적으로 지방이 전체 간 무게의 5% 이상을 초과하는 병적 상태를 의미하는데, 이를 포함하는 간질환은 선진국 40-50대 성인 인구의 사망 원인에서 암 다음으로 심각한 질환으로 인정된다. 선진국을 포함한 주요 국가의 인구 중 30%는 지방간 증상을 보이고 있으며, 이들 중 20%는 간경변증으로 진행하게 되며, 간경변증 환자의 약 절반은 진단 후 10년 이내에 간질환으로 사망한다. Fatty liver refers to a state in which fat is accumulated in liver cells, and medically refers to a pathological condition in which fat exceeds 5% of the total weight of the liver. It is recognized as the second most serious disease after cancer. 30% of the population of major countries, including developed countries, show fatty liver symptoms, and 20% of them progress to cirrhosis, and about half of cirrhosis patients die of liver disease within 10 years after diagnosis.
지방간은 크게 과음으로 인한 알코올성 지방간 질환(alcoholic fatty liver disease)과 비만, 당뇨병, 고지혈증, 약물 등으로 인한 비알코올성 지방간 질환(nonalcoholic fatty liver disease, NAFLD)으로 나눌 수 있다. Fatty liver can be largely divided into alcoholic fatty liver disease caused by excessive drinking and nonalcoholic fatty liver disease (NAFLD) caused by obesity, diabetes, hyperlipidemia, and drugs.
비알코올성 지방간 질환은 과량의 알코올을 섭취하지 않은 환자에서 염증 반응을 동반하지 않는 단순 지방간, 및 이에 의해 진전되며, 간세포의 염증반응, 간섬유화 및 간경화를 포함하는 넓은 범위의 질환을 의미한다. 비알코올성 지방간 질환은 원인에 따라 원발성과 속발성으로 나뉘는데 원발성은 대사 증후군의 특징인 고지혈, 당뇨, 또는 비만 등에 의해 발생되며, 속발성은 영양적 원인(급격한 체중 감소, 기아, 장 우회술), 다양한 약물, 독성 물질(독버섯, 세균 독소), 대사성 원인 및 기타 요인에 의해 발생하는 것으로 알려져 있다. 원발성 요인인 대사증후군의 중요한 특징인 당뇨 및 비만과 관련된 비알코올성 지방간 질환의 발생률은 당뇨 환자의 약 50%, 비만 환자의 약 76%, 비만한 당뇨 환자에서 거의 대부분 발생하는 것으로 알려져 있다. 또한 알라닌 아미노트랜스퍼라제(Alanine aminotransferase, ALT)의 수준이 증가된 당뇨 및 비만 환자에서 생검을 실시한 결과 지방간염의 발생율은 18-36%로 나타내고 있다.Non-alcoholic fatty liver disease refers to simple fatty liver without inflammatory reactions in patients who have not consumed excessive alcohol, and a wide range of diseases that are developed by it and include inflammatory reactions of hepatocytes, liver fibrosis, and liver cirrhosis. Nonalcoholic fatty liver disease is divided into primary and secondary depending on the cause. The primary is caused by hyperlipidemia, diabetes, or obesity, which are characteristic of metabolic syndrome, and the secondary is caused by nutritional causes (rapid weight loss, starvation, intestinal bypass), various drugs, It is known to be caused by toxic substances (poisonous mushrooms, bacterial toxins), metabolic causes, and other factors. It is known that the incidence of non-alcoholic fatty liver disease related to diabetes and obesity, which is an important feature of metabolic syndrome, which is a primary factor, occurs in about 50% of diabetic patients, about 76% of obese patients, and almost all obese diabetic patients. In addition, as a result of biopsies performed in diabetic and obese patients with increased levels of alanine aminotransferase (ALT), the incidence of steatohepatitis has been shown to be 18-36%.
현재까지 비알코올성 지방간 질환에 대한 확립된 치료법은 없는데 이는 비알코올성 지방간 질환이 당뇨병, 비만, 관 상동맥 질환, 생활 습관 등의 다양한 요소와 연관되어 나타나기 때문이다. 몇 가지 당뇨 또는 비만 치료 약제 투여에 의한 지방간 질환에 대한 효과가 보고되었는데 경구용 비만치료제로 사용되고 있는 올리스탓(Orlistat)의 경우 지방간염 환자에서 간의 조직학적 향상을 유도했다는 보고가 있으며, 메트포민(Metformin)은 당뇨를 동반하지 않는 비알코올성 지방간 환자에서 혈중 간 효소 수치 및 간의 괴사성 염증 및 섬유화가 감소시켰다는 보고가 있다. 또한 PPAR(peroxisome proliferator-activated receptor)의 작용물질(agonist)인 티아졸리딘디온(Thiazolidinedione, TZD) 계열의 약물은 간과 근육에서의 지방 축척을 억제하고, 비알코올성 지방간 질환의 동물 모델에서 간에 대해 직접적인 항 섬유화 작용을 나타내는 것으로 보고된 바 있다. To date, there is no established treatment for NAFLD because NAFLD is associated with various factors such as diabetes, obesity, coronary artery disease, and lifestyle. Effects on fatty liver disease caused by administration of several diabetes or obesity treatment drugs have been reported. Orlistat, which is used as an oral obesity treatment, has been reported to induce histological improvement of the liver in patients with steatohepatitis. ) has been reported to reduce blood liver enzyme levels and liver necrotic inflammation and fibrosis in non-alcoholic fatty liver patients without diabetes. In addition, Thiazolidinedione (TZD) drugs, which are agonists of PPAR (peroxisome proliferator-activated receptor), inhibit fat accumulation in the liver and muscle and have a direct effect on the liver in animal models of non-alcoholic fatty liver disease. It has been reported to exhibit anti-fibrotic activity.
그러나 현재 지방간을 약물학적으로 치료하는데 유용한 약제는 거의 없는 상태이며 운동과 식이요법만이 권장되고 있기 때문에 지방간 치료제의 개발이 요구된다.However, there are currently few drugs useful for pharmacologically treating fatty liver, and only exercise and diet are recommended, so the development of a therapeutic agent for fatty liver is required.
본 발명이 해결하고자 하는 과제는 신규한 피롤로 피리미딘 유도체 화합물을 제공하는 것이다.An object to be solved by the present invention is to provide a novel pyrrolopyrimidine derivative compound.
또한, 본 발명은 상기 신규한 피롤로 피리미딘 유도체 화합물의 제조방법을 제공하는 것이다.In addition, the present invention provides a method for preparing the novel pyrrolopyrimidine derivative compound.
또한, 본 발명은 상기 신규 피롤로 피리미딘 유도체 화합물을 유효성분으로 포함하는 대사성 간질환의 예방, 치료 또는 개선용 조성물을 제공하는 것이다.In addition, the present invention is to provide a composition for preventing, treating or improving metabolic liver disease comprising the novel pyrrolopyrimidine derivative compound as an active ingredient.
또한, 본 발명은 상기 신규 피롤로 피리미딘 유도체 화합물을 대상체에게 치료학적 유효량으로 투여하는 단계를 포함하는, 대사성 간질환의 치료방법을 제공하는 것이다.In addition, the present invention provides a method for treating metabolic liver disease, comprising administering the novel pyrrolopyrimidine derivative compound in a therapeutically effective amount to a subject.
본 발명이 해결하고자 하는 과제는 이상에서 언급한 과제(들)로 제한되지 않으며, 언급되지 않은 또 다른 과제(들)는 이하의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.The problem to be solved by the present invention is not limited to the above-mentioned problem (s), and another problem (s) not mentioned will be clearly understood by those skilled in the art from the following description.
상기 과제를 해결하기 위하여, 본 발명은 하기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 제공한다.In order to solve the above problems, the present invention provides a pyrrolopyrimidine derivative compound represented by Formula 1 below, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
[화학식 1][Formula 1]
Figure PCTKR2022011400-appb-img-000001
Figure PCTKR2022011400-appb-img-000001
상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되, In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
또한, 본 발명은 하기 반응식 1에 나타난 바와 같이, 화학식 5로 표시되는 화합물로부터 화학식 1로 표시되는 화합물을 제조하는 단계; 를 포함하는 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물의 제조방법이 제공된다.In addition, as shown in Reaction Scheme 1, the present invention comprises the steps of preparing a compound represented by Formula 1 from a compound represented by Formula 5; A method for preparing a pyrrolopyrimidine derivative compound represented by Formula 1 including a is provided.
[반응식 1][Scheme 1]
Figure PCTKR2022011400-appb-img-000002
Figure PCTKR2022011400-appb-img-000002
상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
또한, 본 발명은 상기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 치료용 약학적 조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing or treating metabolic liver disease comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
또한, 본 발명은 상기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 개선용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for preventing or improving metabolic liver disease comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
또한, 본 발명은 상기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 개선용 동물 사료 조성물을 제공한다.In addition, the present invention provides an animal feed composition for preventing or improving metabolic liver disease, comprising the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
또한, 본 발명은 상기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 대상체에게 치료학적 유효량으로 투여하는 단계를 포함하는, 대사성 간질환의 치료방법을 제공한다.In addition, the present invention provides a method for treating metabolic liver disease, comprising administering a therapeutically effective amount of the compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof to a subject.
본 발명에 따른 신규한 피롤로 피리미딘 유도체 화합물은 지방의 축적을 억제할 수 있어 이를 유효성분으로 포함하여 대사성 간질환, 특히 지방간을 예방, 치료 또는 개선할 수 있는 약학적 조성물, 식품 조성물 또는 사료 조성물로 제공될 수 있다.The novel pyrrolopyrimidine derivative compound according to the present invention can inhibit the accumulation of fat, so a pharmaceutical composition, food composition or feed that can prevent, treat or improve metabolic liver disease, especially fatty liver, by including it as an active ingredient composition may be provided.
본 발명의 효과는 상기한 효과로 한정되는 것은 아니며, 본 발명의 상세한 설명 또는 청구범위에 기재된 발명의 구성으로부터 추론 가능한 모든 효과를 포함하는 것으로 이해되어야 한다.The effects of the present invention are not limited to the above effects, and should be understood to include all effects that can be inferred from the detailed description of the present invention or the configuration of the invention described in the claims.
도 1은 실시예 1 및 2에서 제조한 화합물을 처리 후 HepG2 인간 간세포에서의 지방 방울 변화를 형광으로 나타낸 이미지이다.1 is a fluorescent image of fat droplet change in HepG2 human hepatocytes after treatment with the compounds prepared in Examples 1 and 2.
도 2는 도 1의 이미지를 Gen5 소프트웨어를 이용하여 단위 면적당 nile-red 형광의 평균 세기를 분석한 그래프이다.FIG. 2 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 1 using Gen5 software.
도 3은 실시예 1 및 2에서 제조한 화합물을 처리 후 간오가노이드에서의 지방 방울 변화를 형광으로 나타낸 이미지이다.FIG. 3 is a fluorescence image showing changes in fat droplets in liver organoids after treatment with the compounds prepared in Examples 1 and 2.
도 4는 도 2의 이미지를 Gen5 소프트웨어를 이용하여 단위 면적당 nile-red 형광의 평균 세기를 분석한 그래프이다.FIG. 4 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 2 using Gen5 software.
도 5는 실시예 1에서 제조한 화합물의 타목시펜 유도 지방간 모델에서의 지방간 생성 억제 결과이다.5 is a result of inhibition of fatty liver formation in a tamoxifen-induced fatty liver model by the compound prepared in Example 1.
도 6은 실시예 2에서 제조한 화합물의 타목시펜 유도 지방간 모델에서의 지방간 생성 억제 결과이다.Figure 6 is the compound prepared in Example 2 This is the result of inhibition of fatty liver formation in the tamoxifen-induced fatty liver model.
이하에서, 본 발명을 보다 상세하기 설명한다. Hereinafter, the present invention will be described in more detail.
이하 설명은 발명의 이해를 돕기 위해서 제시하는 것이며, 본 발명이 이하 설명의 내용으로 제한되지 않는다.The following description is presented to aid understanding of the invention, and the present invention is not limited to the contents of the description below.
본 발명의 일 측면에서,In one aspect of the invention,
하기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 입체 이성질체 또는 이의 약학적으로 허용 가능한 염이 제공된다.A pyrrolopyrimidine derivative compound represented by Formula 1 below, an optical stereoisomer thereof, or a pharmaceutically acceptable salt thereof is provided.
[화학식 1][Formula 1]
Figure PCTKR2022011400-appb-img-000003
Figure PCTKR2022011400-appb-img-000003
상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
본 발명에서 상기 “할로젠”은 플루오로, 클로로, 브로모 또는 요오드이며, 바람직하게 플루오로 일 수 있다. In the present invention, the “halogen” may be fluoro, chloro, bromo or iodine, preferably fluoro.
본 발명의 일 구체예에서,In one embodiment of the present invention,
상기 R1은 N-아세틸알라닌, 또는 할로젠으로 치환된 페닐일 수 있다.The R 1 may be N-acetylalanine or phenyl substituted with halogen.
본 발명의 다른 구체예에서,In another embodiment of the invention,
상기 화학식 1로 표시되는 화합물은 하기 화합물 군으로부터 선택되는 어느 하나인 것을 특징으로 하는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염일 수 있다:The compound represented by Formula 1 may be a compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that it is any one selected from the group of compounds below:
4-[(4-플루오로벤젠-1-)설피닐]-7H-피롤로[2,3d]피리미딘4-[(4-fluorobenzene-1-)sulfinyl]-7H-pyrrolo[2,3d]pyrimidine
N-아세틸-S-(7H-피롤로[2,3-d]피리미딘-4-일)-L-시스테인.N-acetyl-S-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-L-cysteine.
본 발명의 화학식 1로 표시되는 화합물은 약학적으로 허용 가능한 염의 형태로 사용할 수 있으며, 염으로는 약학적으로 허용 가능한 유리산(free acid)에 의해 형성된 산 부가염이 유용하다. 산 부가염은 염산, 질산, 인산, 황산, 브롬화수소산, 요드화수소산, 아질산, 아인산 등과 같은 무기산류, 지방족 모노 및 디카르복실레이트, 페닐-치환된 알카노에이트, 하이드록시 알카노에이트 및 알칸디오에이트, 방향족 산류, 지방족 및 방향족 설폰산류 등과 같은 무독성 유기산, 아세트산, 안식향산, 구연산, 젖산, 말레인산, 글루콘산, 메탄설폰산, 4-톨루엔설폰산, 주석산, 푸마르산 등과 같은 유기산으로부터 얻는다. 이러한 약학적으로 무독한 염의 종류로는 설페이트, 피로설페이트, 바이설페이트, 설파이트, 바이설파이트, 니트레이트, 포스페이트, 모노하이드로겐 포스페이트, 다이하이드로겐 포스페이트, 메타포스페이트, 피로포스페이트 클로라이드, 브로마이드, 아이오다이드, 플루오라이드, 아세테이트, 프로피오네이트, 데카노에이트, 카프릴레이트, 아크릴레이트, 포메이트, 이소부티레이트, 카프레이트, 헵타노에이트, 프로피올레이트, 옥살레이트, 말로네이트, 석시네이트, 수베레이트, 세바케이트, 푸마레이트, 말리에이트, 부틴-1,4-디오에이트, 헥산-1,6-디오에이트, 벤조에이트, 클로로벤조에이트, 메틸벤조에이트, 디니트로 벤조에이트, 하이드록시벤조에이트, 메톡시벤조에이트, 프탈레이트, 테레프탈레이트, 벤젠설포네이트, 톨루엔설포네이트, 클로로벤젠설포네이트, 크실렌설포네이트, 페닐아세테이트, 페닐프로피오네이트, 페닐부티레이트, 시트레이트, 락테이트, β-하이드록시부티레이트, 글리콜레이트, 말레이트, 타트레이트, 메탄설포네이트, 프로판설포네이트, 나프탈렌-1-설포네이트, 나프탈렌-2-설포네이트, 만델레이트 등을 포함한다.The compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable salt, and an acid addition salt formed by a pharmaceutically acceptable free acid is useful as the salt. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. It is obtained from non-toxic organic acids such as dioate, aromatic acids, aliphatic and aromatic sulfonic acids, etc., organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, i. Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube rate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.
본 발명에 따른 산 부가염은 통상의 방법으로 제조할 수 있으며, 예를 들면 상기 화학식 1의 피롤로 피리미딘 유도체 화합물을 메탄올, 에탄올, 아세톤, 디클로로메탄, 아세토니트릴 등과 같은 유기용매에 녹이고 유기산 또는 무기산을 가하여 생성된 침전물을 여과, 건조시켜 제조하거나, 용매와 과량의 산을 감압 증류한 후 건조시켜 유기용매 하에서 결정화시켜서 제조할 수 있다.The acid addition salt according to the present invention can be prepared by a conventional method, for example, dissolving the pyrrolo pyrimidine derivative compound of Formula 1 in an organic solvent such as methanol, ethanol, acetone, dichloromethane, acetonitrile, etc., and adding an organic acid or It can be prepared by filtering and drying the precipitate produced by adding an inorganic acid, or by distilling the solvent and excess acid under reduced pressure, drying it, and crystallizing it in an organic solvent.
또한, 염기를 사용하여 약학적으로 허용가능한 금속염을 만들 수 있다. 알칼리 금속 또는 알칼리 토금속 염은 예를 들면 화합물을 과량의 알칼리 금속 수산화물 또는 알칼리 토금속 수산화물 용액 중에 용해하고, 비용해 화합물 염을 여과하고, 여액을 증발, 건조시켜 얻는다. 이때, 금속염으로는 나트륨, 칼륨 또는 칼슘염을 제조하는 것이 제약상 적합하다. 또한, 이에 대응하는 염은 알칼리 금속 또는 알칼리 토금속 염을 적당한 음염(예, 질산은)과 반응시켜 얻는다.In addition, a pharmaceutically acceptable metal salt may be prepared using a base. An alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. In addition, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg, silver nitrate).
나아가, 본 발명은 상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물 및 이의 약학적으로 허용가능한 염뿐만 아니라, 이로부터 제조될 수 있는 용매화물, 광학 이성질체, 수화물 등을 모두 포함한다.Furthermore, the present invention includes not only the pyrrolopyrimidine derivative compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like prepared therefrom.
본 발명의 다른 측면에서,In another aspect of the invention,
하기 반응식 1에 나타난 바와 같이, As shown in Scheme 1 below,
화학식 5로 표시되는 화합물로부터 화학식 1로 표시되는 화합물을 제조하는 단계; 를 포함하는 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물의 제조방법이 제공된다.Preparing a compound represented by Formula 1 from a compound represented by Formula 5; A method for preparing a pyrrolopyrimidine derivative compound represented by Formula 1 including a is provided.
[반응식 1][Scheme 1]
Figure PCTKR2022011400-appb-img-000004
Figure PCTKR2022011400-appb-img-000004
상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
본 발명의 일 구체예에서,In one embodiment of the present invention,
상기 화학식 1의 X가 설피닐인 경우, 상기 화학식 1로 표시되는 화합물이 하기 화학식 1-1로 표시되는 화합물인 것으로 이해될 수 있고,When X in Formula 1 is sulfinyl, it can be understood that the compound represented by Formula 1 is a compound represented by Formula 1-1,
상기 반응식 1로 표시되는 제조방법은 하기 반응식 2에 나타난 바와 같이,The manufacturing method represented by Scheme 1 is as shown in Scheme 2 below,
화학식 5로 표시되는 화합물과 화학식 4로 표시되는 화합물을 반응시켜 화학식 3으로 표시되는 화합물을 제조하는 단계(단계 1); 및 상기 단계 1에서 제조한 화학식 3으로 표시되는 화합물과 옥손(oxone)을 반응시켜 화학식 1-1로 표시되는 화합물을 제조하는 단계(단계 2); 를 포함하는 것일 수 있다.preparing a compound represented by Chemical Formula 3 by reacting a compound represented by Chemical Formula 5 with a compound represented by Chemical Formula 4 (Step 1); and preparing a compound represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone (Step 2); It may contain.
[반응식 2] [Scheme 2]
Figure PCTKR2022011400-appb-img-000005
Figure PCTKR2022011400-appb-img-000005
상기 반응식 2에서, R1a는 치환 또는 비치환된 페닐이되,In Scheme 2, R 1a is substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환된다.The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen.
또한 본 발명의 일 구체예에서,Also in one embodiment of the present invention,
상기 화학식 1의 X가 설파닐인 경우, 상기 화학식 1로 표시되는 화합물이 하기 화학식 1-2로 표시되는 화합물인 것으로 이해될 수 있고,When X in Formula 1 is sulfanyl, it can be understood that the compound represented by Formula 1 is a compound represented by Formula 1-2 below,
상기 반응식 1로 표시되는 제조방법은 하기 반응식 3에 나타난 바와 같이,The manufacturing method represented by Scheme 1 is as shown in Scheme 3 below,
화학식 5로 표시되는 화합물과 화학식 4로 표시되는 화합물을 반응시켜 화학식 3으로 표시되는 화합물을 제조하는 단계(단계 1);preparing a compound represented by Chemical Formula 3 by reacting a compound represented by Chemical Formula 5 with a compound represented by Chemical Formula 4 (Step 1);
상기 단계 1에서 제조한 화학식 3으로 표시되는 화합물과 옥손(oxone)을 반응시켜 화학식 1-1로 표시되는 화합물을 제조하는 단계(단계 2); 및preparing a compound represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone (Step 2); and
상기 단계 2에서 제조한 화학식 1-1로 표시되는 화합물과 화학식 2로 표시되는 화합물을 반응시켜 화학식 1-2로 표시되는 화합물을 제조하는 단계(단계 3); 를 포함하는 것일 수 있다.preparing a compound represented by Chemical Formula 1-2 by reacting the compound represented by Chemical Formula 1-1 prepared in Step 2 with the compound represented by Chemical Formula 2 (Step 3); It may contain.
[반응식 3][Scheme 3]
Figure PCTKR2022011400-appb-img-000006
Figure PCTKR2022011400-appb-img-000006
상기 반응식 3에서, R1a는 치환 또는 비치환된 페닐이되,In Scheme 3, R 1a is substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되며,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
R1b는 N-아세틸알라닌이다.R 1b is N-acetylalanine.
상기 반응식 2 또는 반응식 3으로 표시되는 제조방법에 있어서, 상기 반응식 2 또는 반응식 3의 단계 1은 화학식 5로 표시되는 화합물과 화학식 4로 표시되는 화합물을 반응시켜 화학식 3으로 표시되는 화합물을 제조하는 단계이다.In the preparation method represented by Scheme 2 or Scheme 3, Step 1 of Scheme 2 or Scheme 3 is a step of preparing a compound represented by Formula 3 by reacting a compound represented by Formula 5 with a compound represented by Formula 4. am.
상기 단계 1은 본 발명의 화합물의 모핵구조체인 피롤로 피리미딘에 -S-R1a 치환기를 도입하는 단계로 이해될 수 있고, 상기 단계 1의 반응시간 및 반응 온도는 하기 실시예를 참조하여 해당 분야의 당업자가 용이하게 변경할 수 있는 범위라면 제한 없이 본 발명의 범주에 포함된다. 특히 단계 1에서 제조하고자 하는 화학식 3으로 표시되는 화합물이 제조될 수 있는 조건이면 제한되지 않고 본 발명의 범주내에 포함되는 것으로 이해되어야 한다. 다만 하나의 예시로, 바람직하게, 반응 시간은 5 내지 20시간 수행될 수 있고, 반응 온도는 120 내지 140℃에서 수행될 수 있다. 제한되지 않는 예로, 화학식 5로 표시되는 화합물과 화학식 4로 표시되는 화합물을 트리 에틸아민(triethylamine, TEA) 또는 N,N-디이소프로필에틸아민(DIPEA)과 함께 용매에서 환류시켜 화학식 3으로 표시되는 화합물을 제조할 수 있다. 상기 트리 에틸아민(triethylamine, TEA) 또는 N,N-디이소프로필에틸아민(DIPEA)은 친핵성 방향족 치환반응의 염기로서 사용하였다. Step 1 can be understood as a step of introducing a -SR 1a substituent into pyrrolopyrimidine, which is the parent nucleus structure of the compound of the present invention, and the reaction time and reaction temperature of step 1 can be determined by referring to the following examples. Any range that can be easily changed by those skilled in the art is included in the scope of the present invention without limitation. In particular, it should be understood that any condition under which the compound represented by Formula 3 to be prepared in step 1 can be prepared is not limited and is included within the scope of the present invention. As just one example, preferably, the reaction time may be carried out for 5 to 20 hours, and the reaction temperature may be carried out at 120 to 140 ° C. As a non-limiting example, a compound represented by Formula 5 and a compound represented by Formula 4 are refluxed together with triethylamine (TEA) or N,N-diisopropylethylamine (DIPEA) in a solvent to obtain Formula 3 compounds can be prepared. The triethylamine (TEA) or N,N-diisopropylethylamine (DIPEA) was used as a base for the nucleophilic aromatic substitution reaction.
상기 단계 1에서 사용 가능한 용매로는 H2O, 메탄올, 에탄올, 프로판올, 부탄올, 테트라하이드로퓨란(THF), 디클로로메탄, 톨루엔, 아세토니트릴, 디메틸포름아미드 등을 사용할 수 있고, 바람직하게는 부탄올을 사용할 수 있다.As the solvent usable in step 1, H 2 O, methanol, ethanol, propanol, butanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylformamide, etc. may be used, preferably butanol. can be used
상기 반응식 2 또는 반응식 3으로 표시되는 제조방법에 있어서, 상기 반응식 2 또는 반응식 3의 단계 2는 상기 단계 1에서 제조한 화학식 3으로 표시되는 화합물과 옥손(oxone)을 반응시켜 화학식 1-1로 표시되는 화합물을 제조하는 단계이다.In the preparation method represented by Reaction Scheme 2 or Scheme 3, step 2 of Reaction Scheme 2 or Scheme 3 is represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone. This is the step of preparing a compound to be.
상기 단계 2는 본 발명의 화합물의 모핵구조체인 피롤로 피리미딘에 설피닐 기를 도입하는 단계로 이해될 수 있고, 상기 단계 2의 반응시간 및 반응 온도는 하기 실시예를 참조하여 해당 분야의 당업자가 용이하게 변경할 수 있는 범위라면 제한 없이 본 발명의 범주에 포함된다. 특히 단계 2에서 제조하고자 하는 화학식 1-1로 표시되는 화합물이 제조될 수 있는 조건이면 제한되지 않고 본 발명의 범주내에 포함되는 것으로 이해되어야 한다. 다만 하나의 예시로, 바람직하게, 반응 시간은 5 내지 20시간 수행될 수 있고, 반응 온도는 0 내지 30 ℃에서 수행될 수 있다. 제한되지 않는 예로, 화학식 3으로 표시되는 화합물을 용매에 녹인 후, 또 다른 용매에 녹인 옥손을 혼합하여 반응시켜 화학식 1-1로 표시되는 화합물을 제조할 수 있다. Step 2 can be understood as a step of introducing a sulfinyl group into pyrrolopyrimidine, which is the parent nuclear structure of the compound of the present invention, and the reaction time and reaction temperature of step 2 can be understood by those skilled in the art with reference to the following examples. Any range that can be easily changed is included in the scope of the present invention without limitation. In particular, it should be understood that any condition under which the compound represented by Formula 1-1 to be prepared in step 2 can be prepared is not limited and is included within the scope of the present invention. As just one example, preferably, the reaction time may be carried out for 5 to 20 hours, and the reaction temperature may be carried out at 0 to 30 °C. As a non-limiting example, after dissolving the compound represented by Chemical Formula 3 in a solvent, the compound represented by Chemical Formula 1-1 may be prepared by mixing and reacting with oxone dissolved in another solvent.
이때, 단계 2에서 사용 가능한 용매로는 H2O, 메탄올, 에탄올, 프로판올, 부탄올, 테트라하이드로퓨란(THF), 디클로로메탄, 톨루엔, 아세토니트릴, 디메틸포름아미드 등을 사용할 수 있고, 바람직하게는 H2O, 테트라하이드로퓨란(THF) 또는 이들의 혼합액을 사용할 수 있으며, H2O과 테트라하이드로퓨란(THF)의 혼합하여 사용하는 경우 H2O: 테트라하이드로퓨란(THF)는 1: 2의 비율로 혼합하여 사용할 수 있다.At this time, as a solvent usable in step 2, H 2 O, methanol, ethanol, propanol, butanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylformamide, etc. may be used, preferably H 2 O, tetrahydrofuran (THF) or a mixture thereof can be used, and when using a mixture of H 2 O and tetrahydrofuran (THF), the ratio of H 2 O: tetrahydrofuran (THF) is 1: 2 can be used in combination with
한편, 본 발명은 상기 반응식 2로 표시되는 제조방법에서 화학식 1-1로 표시되는 화합물을 제조하는 것뿐만 아니라, 상기 반응식 3의 단계 3을 수행하여 화학식 1-2로 표시되는 화합물을 제조할 수 있다.On the other hand, the present invention can prepare the compound represented by Chemical Formula 1-2 by performing step 3 of Reaction Scheme 3 as well as preparing the compound represented by Chemical Formula 1-1 in the preparation method represented by Reaction Scheme 2 above. there is.
상기 반응식 3의 단계 3은 상기 단계 2에서 제조한 화학식 1-1로 표시되는 화합물과 화학식 2로 표시되는 화합물을 반응시켜 화학식 1-2로 표시되는 화합물을 제조하는 단계이다.Step 3 of Reaction Scheme 3 is a step for preparing a compound represented by Chemical Formula 1-2 by reacting the compound represented by Chemical Formula 1-1 prepared in Step 2 with the compound represented by Chemical Formula 2.
본 발명에서는 상기 화학식 1-2로 표시되는 화합물은 상기 단계 1에서 화학식 5로 표시되는 화합물과 상기 단계 3의 화학식 2로 표시되는 화합물을 반응시켜 화학식 1-2로 표시되는 화합물을 제조할 수는 있으나, 이 경우 반응이 완전히 전환(conversion)되지 않아 화학식 1-2로 표시되는 화합물의 수득률이 매우 낮다. 따라서, 화학식 1-2로 표시되는 화합물의 수득률을 높이기 위해 단계 1을 거쳐 화학식 5로 표시되는 화합물의 염소를 설파닐(-S-)기로 치환시키고, 단계 2를 거쳐 화학식 3으로 표시되는 화합물의 설파닐(-S-)기를 살피닐(-S=O)기로 치환 시킨 후 단계 3을 거쳐 상기 화학식 1-2로 표시되는 화합물을 제조할 수 있다.In the present invention, the compound represented by Chemical Formula 1-2 can react the compound represented by Chemical Formula 5 in Step 1 with the compound represented by Chemical Formula 2 in Step 3 to prepare the compound represented by Chemical Formula 1-2 However, in this case, the reaction is not completely converted, so the yield of the compound represented by Chemical Formula 1-2 is very low. Therefore, in order to increase the yield of the compound represented by Chemical Formula 1-2, the chlorine of the compound represented by Chemical Formula 5 is substituted with a sulfanyl (-S-) group through Step 1, and the compound represented by Chemical Formula 3 is obtained through Step 2. After substituting the sulfanyl (-S-) group with a salpinyl (-S=O) group, the compound represented by Chemical Formula 1-2 may be prepared through step 3.
상기 단계 3은 본 발명의 화합물의 모핵구조체인 피롤로 피리미딘에 -S-R1b 치환기를 도입하는 단계로 이해될 수 있고, 상기 단계 3의 반응시간 및 반응 온도는 하기 실시예를 참조하여 해당 분야의 당업자가 용이하게 변경할 수 있는 범위라면 제한 없이 본 발명의 범주에 포함된다. 특히 단계 3에서 제조하고자 하는 화학식 1-2로 표시되는 화합물이 제조될 수 있는 조건이면 제한되지 않고 본 발명의 범주내에 포함되는 것으로 이해되어야 한다. 다만 하나의 예시로, 바람직하게, 반응 시간은 5 내지 20시간 수행될 수 있고, 반응 온도는 0 내지 30 ℃에서 수행될 수 있다. 제한되지 않는 예로, 화학식 1-1로 표시되는 화합물을 용매에 녹인 후, 화학식 2로 표시되는 화합물을 첨가하여 반응시켜 화학식 1-2로 표시되는 화합물을 제조할 수 있다.Step 3 can be understood as a step of introducing a -SR 1b substituent into pyrrolopyrimidine, which is the parent nuclear structure of the compound of the present invention, and the reaction time and reaction temperature of step 3 can be determined by referring to the following examples. Any range that can be easily changed by those skilled in the art is included in the scope of the present invention without limitation. In particular, it should be understood that any condition under which the compound represented by Formula 1-2 to be prepared in step 3 can be prepared is not limited and is included within the scope of the present invention. As just one example, preferably, the reaction time may be carried out for 5 to 20 hours, and the reaction temperature may be carried out at 0 to 30 °C. As a non-limiting example, the compound represented by Chemical Formula 1-2 may be prepared by dissolving the compound represented by Chemical Formula 1-1 in a solvent, adding the compound represented by Chemical Formula 2 and reacting the compound represented by Chemical Formula 1-2.
이때, 단계 3에서 사용 가능한 용매로는 H2O, 메탄올, 에탄올, 프로판올, 부탄올, 테트라하이드로퓨란(THF), 디클로로메탄, 톨루엔, 아세토니트릴, 디메틸포름아미드 등을 사용할 수 있고, 바람직하게는 아세토니트릴을 사용할 수 있다.At this time, as a solvent usable in step 3, H 2 O, methanol, ethanol, propanol, butanol, tetrahydrofuran (THF), dichloromethane, toluene, acetonitrile, dimethylformamide, etc. may be used, preferably aceto Nitrile may be used.
한편, 본 발명에서 반응 결과 얻어진 화합물은 유기화학 분야에서 사용되는 일반적인 여과, 세척, 건조 및 농축시키는 단계를 거쳐 단리될 수 있다.Meanwhile, the compound obtained as a result of the reaction in the present invention may be isolated through general filtration, washing, drying, and concentration steps used in the field of organic chemistry.
또한, 본 발명의 다른 측면에서,Also, in another aspect of the present invention,
상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 치료용 약학적 조성물이 제공된다.Provided is a pharmaceutical composition for preventing or treating metabolic liver disease comprising a pyrrolopyrimidine derivative represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
[화학식 1][Formula 1]
Figure PCTKR2022011400-appb-img-000007
Figure PCTKR2022011400-appb-img-000007
상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
여기서, 상기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염은 지방의 축적을 억제하는 유효성분으로 이해될 수 있다.Here, the compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt may be understood as an active ingredient for inhibiting fat accumulation.
상기 대사성 간질환은 알코올성 지방간염 또는 비알코올성 지방간염일 수 있으며, 구체적으로 상기 비알코올성 지방간 질환은 단순 지방간 질환, 영양성 지방간 질환, 기아성 지방간 질환, 비만성 지방간 질환, 당뇨병성 지방간 질환 및 지방간염으로 이루어진 군으로부터 선택되는 1 종 이상인 것일 수 있다.The metabolic liver disease may be alcoholic steatohepatitis or non-alcoholic steatohepatitis, specifically, the non-alcoholic fatty liver disease is simple fatty liver disease, nutritional fatty liver disease, starvation fatty liver disease, obesity fatty liver disease, diabetic fatty liver disease and steatohepatitis. It may be one or more selected from the group consisting of.
상기 대사성 간질환에 있어서, 본 발명의 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 이성질체 또는 약학적으로 허용가능한 염은 질환의 예방, 개선, 치료 등의 의학적으로 유용한 효과를 나타낼 수 있으며, 이는 본원 명세서, 실시예 및 실험예에 의하여 뒷받침되는 바, 이를 유효성분으로 함유되는 약학적 조성물이 제공될 수 있는 것이다. In the metabolic liver disease, the pyrrolopyrimidine derivative compound represented by Formula 1, its isomer or pharmaceutically acceptable salt of the present invention can exhibit medically useful effects such as prevention, improvement, and treatment of the disease, This is supported by the present specification, examples and experimental examples, and a pharmaceutical composition containing it as an active ingredient can be provided.
한편, 본 발명의 약학적 조성물의 사용태양 및 사용방법에 따라 유효성분인 상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염의 함량은 당업자의 선택에 따라 적절히 조절하여 사용될 수 있다.On the other hand, the amount of the active ingredient, the pyrrolopyrimidine derivative compound represented by Formula 1, its optical isomer or its pharmaceutically acceptable salt, according to the mode and method of use of the pharmaceutical composition of the present invention, is determined by those skilled in the art. It can be used by properly adjusting.
본 발명의 일 구체예에서, 상기 약학적 조성물은 상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 전체 조성물의 총 중량에 대하여 0.01 내지 50 중량％, 바람직하게는 0.1 내지 25 중량％, 더 바람직하게는 0.1 내지 10 중량％의 양으로 포함할 수 있다.In one embodiment of the present invention, the pharmaceutical composition comprises 0.01 to 50% by weight of the pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, based on the total weight of the total composition. , preferably 0.1 to 25% by weight, more preferably 0.1 to 10% by weight.
상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염은 상기 약학적 조성물 내에 단독으로 포함될 수 있으며, 또는 그 외 약리학적으로 허용 가능한 담체, 부형제, 희석제 또는 부성분과 함께 포함될 수도 있다.The pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof may be included alone in the pharmaceutical composition, or other pharmaceutically acceptable carriers, excipients, diluents or It may also be included with sub-components.
상기 약학적으로 허용되는 담체, 부형제 또는 희석제의 예로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐 피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유, 덱스트린, 칼슘카보네이트, 프로필렌글리콜, 리퀴드 파라핀 및 생리식염수로 이루어진 군에서 선택된 1종 이상을 들 수 있으나, 이에 한정되는 것은 아니며 통상의 담체, 부형제 또는 희석제 모두 사용 가능하다. 또한, 상기 약학적 조성물은 통상의 충진제, 증량제, 결합제, 붕해제, 항응집제, 윤활제, 습윤제, pH 조절제, 영양제, 비타민, 전해질, 알긴산 및 그의 염, 펙트산 및 그의 염, 보호성 콜로라이드, 글리세린, 향료, 유화제 또는 방부제 등을 추가로 포함할 수 있다.Examples of the pharmaceutically acceptable carrier, excipient or diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , dextrin, calcium carbonate, propylene glycol, liquid paraffin, and at least one selected from the group consisting of physiological saline, but is not limited thereto, and all conventional carriers, excipients, or diluents may be used. In addition, the pharmaceutical composition may include conventional fillers, extenders, binders, disintegrants, anti-agglomerates, lubricants, wetting agents, pH adjusters, nutrients, vitamins, electrolytes, alginic acid and its salts, pectic acid and its salts, protective chlorides, Glycerin, flavors, emulsifiers or preservatives may be further included.
또한, 상기 약학적 조성물은 상기 유효성분 이외에도 대사성 간질환의 치료 또는 예방에 유효한 것으로 공지된 1종 이상의 다른 치료제를 포함하여 동시 또는 이시에 적용되는 병용 요법으로 사용할 수 있다.In addition, the pharmaceutical composition may include one or more other therapeutic agents known to be effective for the treatment or prevention of metabolic liver disease in addition to the active ingredient, and may be used as a combination therapy applied simultaneously or at different times.
상기 약학적 조성물의 투여방법은 경구 또는 비경구 모두 가능하며, 일 예로는 경구, 경피, 피하, 정맥 또는 근육을 포함한 여러 경로를 통해 투여될 수 있다. 또한, 상기 조성물의 제형은 사용방법에 따라 달라질 수 있으며, 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 본 발명이 속하는 기술분야에 잘 알려진 방법을 사용하여 제형화될 수 있다. 일반적으로는, 경구 투여를 위한 고형제제에는 정제(TABLETS), 알약, 연질 또는 경질 캅셀제(CAPSULES), 환제(PILLS), 산제(POWDERS) 및 과립제(GRANULES) 등이 포함되고, 이러한 제제는 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용될 수 있다. 경구를 위한 액상 제제로는 현탁제(SUSTESIONS), 내용액제, 유제(EMULSIONS) 및 시럽제(SYRUPS) 등이 해당되는데, 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제 예를 들면, 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구투여를 위한 형태는 크림(CREAM), 로션제(LOTIONS), 연고제(ONITMENTS), 경고제(PLASTERS), 액제(LIQUIDS AND SOULTIONS), 에어로솔제(AEROSOLS), 유동엑스제(FRUIDEXTRACTS), 엘릭서(ELIXIR), 침제(INFUSIONS), 향낭(SACHET), 패취제(PATCH) 또는 주사제(INJECTIONS) 등의 형태일 수 있으며, 주사용 제형이 될 경우 바람직하게는 등장성 수용액 또는 현탁액의 형태가 될 수 있다.The method of administration of the pharmaceutical composition can be either oral or parenteral, and for example, it can be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular. In addition, the formulation of the composition may vary depending on the method of use, and is formulated using a method well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. It can be. In general, solid preparations for oral administration include tablets (TABLETS), pills, soft or hard capsules (CAPSULES), pills (PILLS), powders (POWDERS) and granules (GRANULES), etc., and these preparations include one or more Excipients, for example, may be prepared by mixing starch, calcium carbonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid formulations for oral use include suspensions, solutions for oral use, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, Sweetening agents, flavoring agents, preservatives, and the like may be included. Forms for parenteral administration include creams, lotions, ointments, PLASTERS, LIQUIDS AND SOULTIONS, aerosols, FRUIDEXTRACTS, and elixirs. It may be in the form of (ELIXIR), INFUSIONS, SACHET, PATCH, or INJECTIONS, and may be in the form of an isotonic aqueous solution or suspension preferably in the case of an injectable formulation. .
상기 약학적 조성물은 멸균제, 방부제, 안정화제, 수화제 또는 유화 촉진제, 삼투압 조절을 위한 염 및/또는 완충제 등의 보조제와, 기타 치료학적으로 유용한 물질을 더 함유할 수 있으며, 통상적인 혼합, 과립화 또는 코팅방법에 따라 제제화할 수 있으며, 이외에도 당해 기술 분야의 공지된 적절한 방법을 사용하여 제형화 할 수 있다.The pharmaceutical composition may further contain adjuvants such as sterilizers, preservatives, stabilizers, hydration agents or emulsification accelerators, salts and/or buffers for osmotic pressure control, and other therapeutically useful substances, and may be conventionally mixed or granulated. It can be formulated according to a coating or coating method, and in addition, it can be formulated using an appropriate method known in the art.
또한, 상기 약학적 조성물의 투여량은 투여방법, 복용자의 연령, 성별, 환자의 중증도, 상태, 체내에서 활성 성분의 흡수도, 불활성율 및 병용되는 약물을 고려하여 결정할 수 있으며, 1회 또는 수회로 나누어 투여할 수 있다. 약학적 조성물의 유효성분으로서 바람직하게는 사람을 비롯한 포유류에게 1일 기준으로 0.001 내지 100 ㎎/㎏ 체중, 바람직하게는 0.01 내지 35㎎/㎏ 체중의 양으로, 1일 1회 또는 분할하여 경구 또는 비경구 경로로 투여될수 있다.In addition, the dosage of the pharmaceutical composition may be determined in consideration of the administration method, the age and sex of the user, the severity of the patient, the condition, the absorption of the active ingredient in the body, the inactivity rate, and the drugs used in combination, once or several times. It can be administered in divided doses. As an active ingredient of the pharmaceutical composition, preferably in an amount of 0.001 to 100 mg/kg body weight, preferably 0.01 to 35 mg/kg body weight, on a daily basis to mammals including humans, once a day or in divided doses orally or It can be administered by the parenteral route.
본 발명의 일 구체예에서, 상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 대상체에게 치료학적 유효량으로 투여하는 단계를 포함하는, 대사성 간질환의 치료방법을 제공한다.In one embodiment of the present invention, the treatment of metabolic liver disease, comprising administering to a subject a therapeutically effective amount of a pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof. provides a treatment method.
바람직하게는 상기 치료방법은 상기 투여 단계 이전에 상기 대사성 간질환의 예방 또는 치료를 필요로 하는 환자를 확인하는 단계를 추가로 포함할 수 있다. Preferably, the treatment method may further include a step of identifying a patient in need of prevention or treatment of the metabolic liver disease prior to the administration step.
상기 "치료학적 유효량"은 대사성 간질환의 예방 또는 치료에 효과적인, 포유류에 대한 유효 성분의 양을 의미하며, 상기 치료학적 유효량은 질환의 종류, 질환의 중증도, 조성물에 함유된 유효 성분 및 다른 성분의 종류 및 함량, 제형의 종류 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 혈중 청소율, 치료 기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있으나, 바람직하게는 상술한 바와 같이, 1일 기준으로 0.001 내지 100 ㎎/㎏ 체중, 바람직하게는 0.01 내지 35 ㎎/㎏ 체중의 양으로, 1일 1회 또는 분할하여 경구 또는 비경구 경로로 투여할 수 있다.The "therapeutically effective amount" means an amount of an active ingredient for mammals effective for preventing or treating metabolic liver disease, and the therapeutically effective amount is the type of disease, the severity of the disease, the active ingredient and other ingredients contained in the composition It can be adjusted according to various factors, including the type and content of the formulation and the patient's age, weight, general health condition, sex and diet, administration time, administration route and blood clearance of the composition, treatment period, and concurrently used drugs. However, preferably, as described above, 0.001 to 100 mg / kg body weight per day, preferably 0.01 to 35 mg / kg body weight, administered once a day or divided by oral or parenteral route can do.
상기 “대상체”는 인간 또는 비-인간인 영장류, 생쥐(mouse), 개, 고양이, 말, 소 등의 포유류를 의미할 수 있으나, 이에 한정되는 것은 아니다.The “subject” may refer to mammals such as humans or non-human primates, mice, dogs, cats, horses, and cows, but is not limited thereto.
또한 본 발명의 다른 측면에서,Also, in another aspect of the present invention,
상기 화학식 1로 표시되는 화합물, 이의 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 개선용 식품 조성물이 제공된다. Provided is a food composition for preventing or improving metabolic liver disease comprising the compound represented by Formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
본 발명의 일 구체예에서, 상기 식품 조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food), 식품 첨가제(food additives) 또는 사료 등의 모든 형태를 포함하는 의미이며, 인간 또는 가축을 비롯한 동물을 취식대상으로 한다. 상기 유형의 식품 조성물은 당 업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다.In one embodiment of the present invention, the food composition is meant to include all forms of functional food, nutritional supplements, health food, food additives, or feed. , humans or animals, including livestock, are intended as food. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
상기 유형의 식품 조성물은 당업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다. 일반 식품으로는 이에 한정되지 않지만 음료(알콜성 음료 포함), 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마아말레이드 등), 어류, 육류 및 그 가공식품(예: 햄, 소시지 콘비이프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게이트, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품(예: 버터, 치이즈 등), 식용식물 유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등에 본 발명의 상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 첨가하여 제조할 수 있다. 또한, 영양보조제로는 이에 한정되지 않지만 캡슐, 타블렛, 환 등에 본 발명 화학식 1의 화합물을 첨가하여 제조할 수 있다. 또한, 건강기능식품으로는 이에 한정되지 않지만 예를 들면, 본 발명의 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 차, 쥬스 및 드링크의 형태로 제조하여 음용(건강음료)할 수 있도록 액상화, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. Food compositions of this type can be prepared in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruits, bottled products, jams, marmalades, etc.), fish, meat and their processed foods (e.g. ham, sausages) Corned beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , vegetable protein, retort food, frozen food, various seasonings (e.g., soybean paste, soy sauce, sauce, etc.), etc. The pyrrolopyrimidine derivative compound represented by Formula 1 of the present invention, its optical isomer or its pharmaceutically acceptable salt It can be prepared by adding In addition, nutritional supplements are not limited thereto, but may be prepared by adding the compound of Formula 1 of the present invention to capsules, tablets, pills, etc. In addition, health functional foods are not limited thereto, but, for example, the pyrrolopyrimidine derivative compound represented by Formula 1 of the present invention, its optical isomer or its pharmaceutically acceptable salt in the form of tea, juice and drink It can be consumed by liquefying, granulating, encapsulating, and powdering so that it can be manufactured and consumed (health drink).
본 발명의 건강식품은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산, 펙트산의 염, 알긴산, 알긴산의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올 또는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강식품은 천연 과일주스, 과일주스 음료, 또는 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물 100 중량부당 0.01 ~ 0.1 중량부의 범위에서 선택되는 것이 일반적이다.The health food of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain alcohol or a carbonating agent, and the like. In addition, the health food of the present invention may contain fruit flesh for producing natural fruit juice, fruit juice beverage, or vegetable beverage. These components may be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.
또한 본 발명의 다른 측면에서,Also, in another aspect of the present invention,
상기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 개선용 동물 사료 조성물이 제공된다.An animal feed composition for preventing or improving metabolic liver disease comprising a pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient is provided.
본 발명에서 용어 '사료'는, 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분으로, 본 발명에 따른 대사성 간질환의 예방 또는 개선용 조성물을 유효성분으로 포함하는 사료는 당업계에 공지된 다양한 형태의 사료로 제조가능하며, 바람직하게는 농후 사료, 조사료 및/또는 특수사료가 포함될 수 있으나, 이로 한정되지 않는다.In the present invention, the term 'feed' refers to any natural or artificial diet, one-meal meal, etc., or a component of the one-meal meal, suitable for or suitable for animals to eat, ingest, and digest, and to prevent metabolic liver disease according to the present invention. Alternatively, the feed containing the composition for improvement as an active ingredient can be prepared from various types of feed known in the art, and preferably includes concentrated feed, roughage and/or special feed, but is not limited thereto.
본 발명에서 용어, '사료첨가제'는 영양소 보충 및 체중감소 예방, 사료 내 섬유소의 소화 이용성 증진, 유질개선, 번식장애 예방 및 수태율 향상, 하절기 고온 스트레스 예방 등 다양한 효과를 목적으로 사료에 첨가하는 물질을 포함한다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당하며, 탄산수소나트륨, 벤토나이트(bentonite), 산화마그네슘, 복합광물질 등의 광물질제제, 아연, 구리, 코발트, 셀레늄 등의 미량 광물질인 미네랄제제, 케로틴, 비타민 A D, E, 니코틴산, 비타민 B 복합체 등의 비타민제, 메티오닌, 라이신 등의 보호아미노산제, 지방산 칼슘염 등의 보호지방산제, 생균제(유산균제), 효모배양물, 곰팡이 발효물 등의 생균, 효모제 등이 추가로 포함될 수 있다.In the present invention, the term 'feed additive' is a substance added to feed for various purposes, such as supplementation of nutrients and prevention of weight loss, enhancement of digestibility of fiber in feed, improvement of oil quality, prevention of reproductive disorders and improvement of conception rate, and prevention of high-temperature stress in summer. includes The feed additive of the present invention corresponds to supplementary feed under the Feed Control Act, and includes mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals, mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium, and kerotene. , vitamins such as vitamins A, D, E, nicotinic acid, and vitamin B complex, protective amino acids such as methionine and lysine, protected fatty acids such as calcium salts of fatty acids, probiotics (lactic acid bacteria), yeast cultures, live bacteria such as mold fermented products, Yeast agents and the like may be further included.
농후사료에는 밀, 귀리, 옥수수 등의 곡류를 포함하는 종자열매류, 곡물을 정제하고 얻는 부산물로서 쌀겨, 밀기울, 보릿겨 등을 포함하는 겨류, 콩, 유체, 깨, 아마인, 코코야자 등을 채유하고 얻는 부산물인 깻묵류와 고구마, 감자 등에서 녹말을 뺀 나머지인 녹말찌꺼기의 주성분인 잔존녹말질류 등의 찌꺼기류, 어분, 물고기찌꺼기, 어류에서 얻은 신선한 액상물을 농축시킨 것인 피시솔루블(fish soluble), 육분(肉粉), 혈분, 우모분, 탈지분유, 우유에서 치즈, 탈지유에서 카제인을 제조할 때의 잔액인 훼이(whey)를 건조한 건조훼이 등의 동물질사료, 효모, 클로렐라, 해조류가 있으나 이에 제한되지 않는다.Concentrated feed includes seed fruits including grains such as wheat, oats, and corn, bran including rice bran, wheat bran, and barley bran as by-products obtained after refining grains, soybeans, fluids, sesame seeds, linseed, and coco palm oil. Fish soluble, which is a condensed fresh liquid obtained from fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal, fish meal soluble), meat meal, blood meal, feather meal, skim milk powder, cheese from milk, dried whey, which is the balance when casein is produced from skim milk, and animal feed such as yeast, chlorella, and seaweed. Not limited to this.
조사료에는 야초, 목초, 풋베기 등의 생초(生草)사료, 사료용 순무, 사료용 비트, 순무의 일종인 루터베어거 등의 뿌리채소류, 생초, 풋베기작물, 곡실(穀實) 등을 사일로에 채워 놓고 젖산발효시킨 저장사료인 사일리지(silage), 야초, 목초를 베어 건조시킨 건초, 종축용(種畜用) 작물의 짚, 콩과 식물의 나뭇잎이 있으며, 이에 제한되지 않는다. 특수사료에는 굴껍데기, 암염 등의 미네랄 사료, 요소나 그 유도체인 디우레이드이소부탄 등의 요소사료, 천연사료원료만을 배합했을 때 부족하기 쉬운 성분을 보충하거나, 사료의 저장성을 높이기 위해서 배합사료에 미량으로 첨가하는 물질인 사료첨가물, 식이보조제가 있으나 이에 제한되지 않는다.Forage, raw grass feed such as wild grass, grass, and green cutting, turnip for feed, beet for feed, root vegetables such as Lutherbearer, a kind of turnip, raw grass, green crops, grain, etc. are put into silos. Silage (silage), which is a stored feed filled and fermented with lactic acid, grass, hay dried by cutting grass, straw of breeding crops, and leaves of leguminous plants, but are not limited thereto. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea or its derivative, diureide isobutane, and supplements for ingredients that tend to be insufficient when only natural feed ingredients are mixed, or formulated feeds to improve the storage life of feeds. There are feed additives and dietary supplements, which are substances added in small amounts, but are not limited thereto.
본 발명에 따른 상기 대사성 간질환의 예방 또는 개선용 사료 첨가제는 당업계에 공지된 다양한 사료 제조방법에 따라 적절한 유효 농도 범위에서 본 발명의 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 첨가하여 제조 가능하다.The feed additive for preventing or improving metabolic liver disease according to the present invention is a pyrrolopyrimidine derivative compound represented by Formula 1 of the present invention, an optical isomer thereof, in an appropriate effective concentration range according to various feed manufacturing methods known in the art Or it can be prepared by adding a pharmaceutically acceptable salt thereof.
중복된 기재를 피하기 위해서, 별도의 언급이 없는 한, 화학식 1 및 약학적 조성물 부분에서 기술한 본 발명 각 구성에 대한 정의 및 설명은 식품 조성물, 사료 조성물에 그대로 준용된다.In order to avoid redundant description, the definitions and descriptions of each component of the present invention described in Formula 1 and the pharmaceutical composition section are applied mutatis mutandis to food compositions and feed compositions, unless otherwise specified.
이하에서, 첨부된 도면을 참조하여 실시예들을 상세하게 설명한다. 그러나, 실시예들에는 다양한 변경이 가해질 수 있어서 특허출원의 권리 범위가 이러한 실시예들에 의해 제한되거나 한정되는 것은 아니다. 실시예들에 대한 모든 변경, 균등물 내지 대체물이 권리 범위에 포함되는 것으로 이해되어야 한다.Hereinafter, embodiments will be described in detail with reference to the accompanying drawings. However, since various changes can be made to the embodiments, the scope of the patent application is not limited or limited by these embodiments. It should be understood that all changes, equivalents or substitutes to the embodiments are included within the scope of rights.
<실시예><Example>
실시예 1. 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘의 제조Example 1. Preparation of 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d]pyrimidine
Figure PCTKR2022011400-appb-img-000008
Figure PCTKR2022011400-appb-img-000008
단계 1: 4-((4-플루오로 페닐)싸이오)-7H-피롤로 [2,3-d] 피리미딘의 제조Step 1: Preparation of 4-((4-fluorophenyl)thio)-7H-pyrrolo[2,3-d]pyrimidine
4-클로로-7-H-피롤로[2,3-d]피리미딘 (1.0g, 6.512mmol)을 4-플루오로-티오 페놀 (0.73 mL, 6.837 mmol) 및 트리 에틸아민 (3.66 mL, 26.047 mmol)과 함께 1- 부탄올(50 mL)에서 16시간 동안 환류시켰다. 휘발성 물질을 진공에서 증발시키고 잔류물을 컬럼 크로마토그래피 (CH2Cl2 / MeOH 9 : 1)로 정제하여 4-((4-플루오로 페닐)싸이오)-7H-피롤로 [2,3-d] 피리미딘 을 1.48 g (92.7 %) 수득하였다. 4-Chloro-7-H-pyrrolo[2,3-d]pyrimidine (1.0 g, 6.512 mmol) was added to 4-fluoro-thiophenol (0.73 mL, 6.837 mmol) and triethylamine (3.66 mL, 26.047 mL). mmol) in 1-butanol (50 mL) for 16 hours. The volatiles were evaporated in vacuo and the residue purified by column chromatography (CH 2 Cl 2 / MeOH 9:1) to give 4-((4-fluorophenyl)thio)-7H-pyrrolo[2,3- d] 1.48 g (92.7%) of pyrimidine.
1H NMR (400MHz, DMSO-d6) δ 6.12 (d, 1H, 3 J(H,H) ) 3.5 Hz, HC=CH), 7.31-7.41 (2H, 4-F-Ph), 7.48 (d, 1H, 3 J (H,H) ) 3.5 Hz, HC=CH), 7.66-7.75 (2H, 4-F-Ph), 8.47 (s, 1H), 12.24 (s, 1H, NH).1H NMR (400MHz, DMSO-d6) δ 6.12 (d, 1H, 3 J(H,H) ) 3.5 Hz, HC=CH), 7.31-7.41 (2H, 4-F-Ph), 7.48 (d, 1H , 3 J (H,H)) 3.5 Hz, HC=CH), 7.66-7.75 (2H, 4-F-Ph), 8.47 (s, 1H), 12.24 (s, 1H, NH).
단계 2: 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘의 제조Step 2: Preparation of 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d]pyrimidine
상기 단계 1에서 제조한 4-((4-플루오로 페닐)싸이오)-7H-피롤로 [2,3-d] 피리미딘 (1.48 g, 6.034 mmol)를 차가운 THF (0 ℃, 50 mL)에 녹인 후, 5 ℃ 이하의 반응 온도를 유지하면서 물에 녹인 황산수소칼륨 (Oxone, 1.947 g, 6.336 mmol, 100 mL)을 추가하였다. 생성된 화합물을 실온에서 16시간 동안 교반하고, 물로 희석한 후, 100mL의 에틸아세테이트로 2회 추출하였다. 생성물을 염수로 세척하고, 황산 나트륨(Na2SO4)으로 건조시키고, 여과 및 농축하여 화합물을 수득하였다. 수득한 화합물을 에틸아세테이트/다이클로로메테인 을 사용하여 실리카겔 컬럼 크로마토그래피로 정제하여 목적하는 순수한 화합물인 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘 (700 mg / 44.4 %)을 수득하였다.4-((4-fluorophenyl)thio)-7H-pyrrolo[2,3-d] pyrimidine (1.48 g, 6.034 mmol) prepared in step 1 was added to cold THF (0 °C, 50 mL). After dissolving, potassium hydrogen sulfate dissolved in water (Oxone, 1.947 g, 6.336 mmol, 100 mL) was added while maintaining the reaction temperature below 5 °C. The resulting compound was stirred at room temperature for 16 hours, diluted with water, and extracted twice with 100 mL of ethyl acetate. The product was washed with brine, dried over sodium sulfate (Na 2 SO 4 ), filtered and concentrated to give the compound. The obtained compound was purified by silica gel column chromatography using ethyl acetate/dichloromethane to obtain the desired pure compound, 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d ] pyrimidine (700 mg / 44.4%).
1H NMR (400MHz, DMSO) δ 12.82 (s, 1H), 8.75 (s, 1H), 7.92-7.88 (m, 2H), 7.75 (dd, J = 3.6 Hz, 1H), 7.43-7.37 (m, 2H), 7.09 (dd, J = 3.5, 1.7 Hz, 1H).1H NMR (400MHz, DMSO) δ 12.82 (s, 1H), 8.75 (s, 1H), 7.92-7.88 (m, 2H), 7.75 (dd, J = 3.6 Hz, 1H), 7.43-7.37 (m, 2H) ), 7.09 (dd, J = 3.5, 1.7 Hz, 1H).
실시예 2. 4N-아세틸-S-(7H-피롤로 [2,3-d] 피리미딘-4-일)-L-시스테인의 제조Example 2. Preparation of 4N-acetyl-S-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-L-cysteine
Figure PCTKR2022011400-appb-img-000009
Figure PCTKR2022011400-appb-img-000009
단계 1: 4-((4-플루오로 페닐)싸이오)-7H-피롤로 [2,3-d] 피리미딘의 제조Step 1: Preparation of 4-((4-fluorophenyl)thio)-7H-pyrrolo[2,3-d]pyrimidine
4-클로로-7-H-피롤로[2,3-d]피리미딘 (1.0g, 6.512mmol)을 4-플루오로-티오 페놀 (0.73 mL, 6.837 mmol) 및 트리 에틸아민 (3.66 mL, 26.047 mmol)과 함께 1- 부탄올(50 mL)에서 16시간 동안 환류시켰다. 휘발성 물질을 진공에서 증발시키고 잔류물을 컬럼 크로마토그래피 (CH2Cl2 / MeOH 9 : 1)로 정제하여 4-((4-플루오로 페닐)싸이오)-7H-피롤로 [2,3-d] 피리미딘 을 1.48 g (92.7 %) 수득하였다. 4-Chloro-7-H-pyrrolo[2,3-d]pyrimidine (1.0 g, 6.512 mmol) was added to 4-fluoro-thiophenol (0.73 mL, 6.837 mmol) and triethylamine (3.66 mL, 26.047 mL). mmol) in 1-butanol (50 mL) for 16 hours. The volatiles were evaporated in vacuo and the residue purified by column chromatography (CH 2 Cl 2 / MeOH 9:1) to give 4-((4-fluorophenyl)thio)-7H-pyrrolo[2,3- d] 1.48 g (92.7%) of pyrimidine.
1H NMR (400MHz, DMSO-d6) δ 6.12 (d, 1H, 3 J(H,H) ) 3.5 Hz, HC=CH), 7.31-7.41 (2H, 4-F-Ph), 7.48 (d, 1H, 3 J (H,H) ) 3.5 Hz, HC=CH), 7.66-7.75 (2H, 4-F-Ph), 8.47 (s, 1H), 12.24 (s, 1H, NH).1H NMR (400MHz, DMSO-d6) δ 6.12 (d, 1H, 3 J(H,H) ) 3.5 Hz, HC=CH), 7.31-7.41 (2H, 4-F-Ph), 7.48 (d, 1H , 3 J (H,H)) 3.5 Hz, HC=CH), 7.66-7.75 (2H, 4-F-Ph), 8.47 (s, 1H), 12.24 (s, 1H, NH).
단계 2: 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘의 제조Step 2: Preparation of 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d]pyrimidine
상기 단계 1에서 제조한 4-((4-플루오로 페닐)싸이오)-7H-피롤로 [2,3-d] 피리미딘 (1.48 g, 6.034 mmol)를 차가운 THF (0 ℃, 50 mL)에 녹인 후, 5 ℃ 이하의 반응 온도를 유지하면서 물에 녹인 황산수소칼륨 (Oxone, 1.947 g, 6.336 mmol, 100 mL)을 추가하였다. 생성된 화합물을 실온에서 16시간 동안 교반하고, 물로 희석한 후, 100mL의 에틸아세테이트로 2회 추출하였다. 생성물을 염수로 세척하고, 황산 나트륨(Na2SO4)으로 건조시키고, 여과 및 농축하여 화합물을 수득하였다. 수득한 화합물을 에틸아세테이트/다이클로로메테인 을 사용하여 실리카겔 컬럼 크로마토그래피로 정제하여 목적하는 순수한 화합물인 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘 (700 mg / 44.4 %)을 수득하였다.4-((4-fluorophenyl)thio)-7H-pyrrolo[2,3-d] pyrimidine (1.48 g, 6.034 mmol) prepared in step 1 was added to cold THF (0 °C, 50 mL). After dissolving, potassium hydrogen sulfate dissolved in water (Oxone, 1.947 g, 6.336 mmol, 100 mL) was added while maintaining the reaction temperature below 5 °C. The resulting compound was stirred at room temperature for 16 hours, diluted with water, and extracted twice with 100 mL of ethyl acetate. The product was washed with brine, dried over sodium sulfate (Na 2 SO 4 ), filtered and concentrated to give the compound. The obtained compound was purified by silica gel column chromatography using ethyl acetate/dichloromethane to obtain the desired pure compound, 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d ] pyrimidine (700 mg / 44.4%).
1H NMR (400MHz, DMSO) δ 12.82 (s, 1H), 8.75 (s, 1H), 7.92-7.88 (m, 2H), 7.75 (dd, J = 3.6 Hz, 1H), 7.43-7.37 (m, 2H), 7.09 (dd, J = 3.5, 1.7 Hz, 1H).1H NMR (400MHz, DMSO) δ 12.82 (s, 1H), 8.75 (s, 1H), 7.92-7.88 (m, 2H), 7.75 (dd, J = 3.6 Hz, 1H), 7.43-7.37 (m, 2H) ), 7.09 (dd, J = 3.5, 1.7 Hz, 1H).
단계 3: N-아세틸-S-(7H-피롤로 [2,3-d] 피리미딘-4-일)-L-시스테인의 제조Step 3: Preparation of N-acetyl-S-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-L-cysteine
상기 단계 2에서 제조한 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘(424 mg, 1.623 mmol)이 첨가된 아세토나이트릴 (15mL) 용액에 N-아세틸-L-시스테인 (529.66 mg, 3.246 mmol)을 첨가하고, 실온에서 16 시간 동안 교반 하였다. 그 후, 혼합물을 진공에서 농축시켰다. 농축시킨 혼합물을 에틸아세테이트/헥산/메탄올을 사용하여 실리카겔 칼럼 크로마토그래피로 정제하여 목적하는 순수한 화합물인 N-아세틸-S-(7H-피롤로 [2,3-d] 피리미딘-4-일)-L-시스테인 (181 mg, 39.8 %)을 수득하였다. Acetonitrile (15mL) solution to which 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d] pyrimidine (424 mg, 1.623 mmol) prepared in step 2 was added N-acetyl-L-cysteine (529.66 mg, 3.246 mmol) was added thereto, and the mixture was stirred at room temperature for 16 hours. The mixture was then concentrated in vacuo. The concentrated mixture was purified by silica gel column chromatography using ethyl acetate/hexane/methanol to obtain N-acetyl-S-(7H-pyrrolo[2,3-d]pyrimidin-4-yl) as the desired pure compound. -L-cysteine (181 mg, 39.8%) was obtained.
1H-NMR (CDCl3, 400 MHz) δ 2.05 (3H, s), 3.74 (1H, q), 3.852 (1H, q), 4.80 (1H, m) 6.12 (d, 1H), 7.4 (d, 1H), 8.47 (s, 1H), 8.91 (1H, br-s), 12.24 (s, 1H, NH).1H-NMR (CDCl3, 400 MHz) δ 2.05 (3H, s), 3.74 (1H, q), 3.852 (1H, q), 4.80 (1H, m) 6.12 (d, 1H), 7.4 (d, 1H) , 8.47 (s, 1H), 8.91 (1H, br-s), 12.24 (s, 1H, NH).
<실험예> 지방간 억제 효능 평가<Experimental Example> Evaluation of fatty liver inhibitory efficacy
본 발명에 따른 신규한 피롤로 피리미딘 유도체 화합물의 지방간 억제 효능을 평가하기 위해, 지방방울(lipid droplet)을 생성 억제를 다음과 같이 실험하였다.In order to evaluate the fatty liver inhibitory effect of the novel pyrrolopyrimidine derivative compound according to the present invention, inhibition of lipid droplet production was tested as follows.
실험예 1. 간세포에서의 지방간 억제 효과Experimental Example 1. Fatty liver inhibitory effect in hepatocytes
HepG2 세포에 Oleic acid와 palmitic acid으로 지방증을 유도한 후, 상기 실시예 1에서 제조한 4-((4-플루오로 페닐)설피닐)-7H-피롤로 [2,3-d] 피리미딘 및 상기 실시예 2에서 제조한 N-아세틸-S-(7H-피롤로 [2,3-d] 피리미딘-4-일)-L-시스테인을 각각 처리하여 세포 내 지방량에 미치는 영향을 관찰하였다.After inducing steatosis in HepG2 cells with oleic acid and palmitic acid, 4-((4-fluorophenyl)sulfinyl)-7H-pyrrolo[2,3-d]pyrimidine and N-acetyl-S-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-L-cysteine prepared in Example 2 was treated, respectively, and the effect on intracellular fat content was observed.
구체적으로, HepG2 세포는 DMEM 배지를 사용하였고, 배지에 10% FBS(Fetal Bovine Serum)와 100μg/mL 항생제를 첨가하였다. 세포는 24시간 동안 5% CO2를 유지하는 37℃ 배양기에서 배양하고, 2-3일마다 70-80% 정도 성장하였을 때, PBS(Phosohate buffer solution)로 세척하고, Trypsin-EDTA(Gibco, USA)을 처리하여 계대 배양하였다. HepG2 세포를 5 x 104 개/웰로 24웰 플레이트에 준비하고 24시간 동안 5% CO2를 유지하는 37℃배양기에서 안정화시켰다. 지방증을 유도하기 위해 Oleic acid와 Palmitic acid를 2:1비율로 최종 1mM 농도가 되도록 처리하였다.Specifically, HepG2 cells used DMEM medium, and 10% FBS (Fetal Bovine Serum) and 100 μg/mL antibiotics were added to the medium. The cells were cultured in a 37°C incubator maintained at 5% CO 2 for 24 hours, and when they reached 70-80% growth every 2-3 days, they were washed with PBS (Phosohate buffer solution), and Trypsin-EDTA (Gibco, USA). ) and subcultured. HepG2 cells were prepared in a 24-well plate at 5 x 10 4 cells/well and stabilized in a 37°C incubator maintained at 5% CO 2 for 24 hours. To induce steatosis, oleic acid and palmitic acid were treated at a ratio of 2:1 to a final concentration of 1 mM.
실시예 1 및 2에서 제조한 화합물을 함유한 배지에서 24시간동안 배양하고, 15분간 4% paraformaldehyde로 고정하였다. PBS(Phosohate buffer solution)로 세척한 뒤, 5분간 Nile red(1μg/mL)로 처리하여 세포내 중성 지방을 염색하고, DAPI를 처리하여 세포 내 핵을 염색하였다. 지방간의 생성 정도를 측정하기 위하여 Lionheart FX Automated Microscope (BIOTEK)로 촬영하고, Gen5 software로 정량화 하였으며, 그 결과를 도 1 및 도 2에 나타내었다.Incubated for 24 hours in a medium containing the compounds prepared in Examples 1 and 2, and fixed with 4% paraformaldehyde for 15 minutes. After washing with PBS (Phosohate buffer solution), intracellular neutral fat was stained by treatment with Nile red (1 μg/mL) for 5 minutes, and intracellular nuclei were stained by treatment with DAPI. In order to measure the degree of fatty liver production, it was photographed with a Lionheart FX Automated Microscope (BIOTEK) and quantified with Gen5 software, and the results are shown in FIGS. 1 and 2.
도 1은 실험예 1에 따라 실시예 1 및 2에서 제조한 화합물을 처리 후 HepG2 인간 간세포에서의 지방 방울 변화를 형광으로 나타낸 이미지로서, 실시예 1 및 2에서 제조한 화합물의 농도가 증가함에 따라 지방산 혼합액에 의해 증가한 지방 방울의 축적을 감소시킴을 확인할 수 있다.1 is an image showing the change in fat droplets in HepG2 human hepatocytes by fluorescence after treatment with the compounds prepared in Examples 1 and 2 according to Experimental Example 1. As the concentration of the compound prepared in Examples 1 and 2 increases, It can be confirmed that the accumulation of fat droplets increased by the fatty acid mixture is reduced.
도 2는 상기 도 1의 이미지를 Gen5 소프트웨어를 이용하여 단위 면적당 nile-red 형광의 평균 세기를 분석한 그래프이다. 지방 염색 결과 Nile red 형광 세기가 지방증 유도군에 비해 실시예 1에 따라 제조한 화합물 1 μM 농도에서 74% 억제되고, 실시예 2에 따라 제조한 화합물 5 μM 농도에서 70% 감소하는 것을 확인할 수 있었다. FIG. 2 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 1 using Gen5 software. As a result of fat staining, it was confirmed that Nile red fluorescence intensity was suppressed by 74% at 1 μM concentration of the compound prepared according to Example 1 and decreased by 70% at 5 μM concentration of the compound prepared according to Example 2 compared to the steatosis induction group. .
실험예 2. 간오가노이드를 이용한 지방간 억제 효능Experimental Example 2. Fatty liver inhibitory effect using liver organoids
본 발명의 실시예 1 및 2에서 제조한 화합물의 지방간 억제 효능을 평가하기 위하여 인간 유래의 간오가노이드에서의 nile-red를 이용한 지방 방울 (lipid droplet) 염색 분석법을 이용하여 실험을 수행하였다.In order to evaluate the fatty liver inhibitory effect of the compounds prepared in Examples 1 and 2 of the present invention, experiments were performed using a lipid droplet staining assay using nile-red in human-derived liver organoids.
인간 유래의 간오가노이를 일반 성장배지에서 배양하였다. 성장배지는 advanced DMEF/F-12 배지에 1% penicilin-streptomycin, 1% Glutamax, 10 mM HEPES, 1x N2, 1x B27 (w/o vitA), 1% insulin-Transferrin-Selenium, 50 ng/mL EGF, 25 ng/mL HGF, 10 ng/mL GFG-basic, 10 ng/mL Oncostatin M, 5 μM A83-01, 10 μM Forskolin, 1 mM n-acetylcysteine, 10 nM Gastrin, 10 mM Nicotinamide, 100 nM Dexamethasone을 포함하도록 제조하였다. 간오가노이드는 2-3일 마다 성장배지를 교체하며, 37℃, 5% CO2의 습윤화된 배양기에서 적응시켜 7일간 배양하였다. Human-derived liver organoids were cultured in a general growth medium. The growth medium was advanced DMEF/F-12 medium containing 1% penicilin-streptomycin, 1% Glutamax, 10 mM HEPES, 1x N2, 1x B27 (w/o vitA), 1% insulin-Transferrin-Selenium, and 50 ng/mL EGF. , 25 ng/mL HGF, 10 ng/mL GFG-basic, 10 ng/mL Oncostatin M, 5 μM A83-01, 10 μM Forskolin, 1 mM n-acetylcysteine, 10 nM Gastrin, 10 mM Nicotinamide, and 100 nM Dexamethasone made to contain. Liver organoids were cultured for 7 days by changing the growth medium every 2-3 days and adapting them in a humidified incubator at 37°C and 5% CO 2 .
본 실험예에서 간오가노이드는 1 mL 마이크로파이펫을 이용하여 10~15회 파이펫팅을 진행하여 단편화 한 후, 단편의 개수가 100~200개가 되도록 matrigel을 이용하여 희석한후, ultra Low binding 96-웰에 3일간 배양하였다. In this experimental example, hepatic organoids were fragmented by pipetting 10 to 15 times using a 1 mL micropipette, diluted with matrigel so that the number of fragments was 100 to 200, and ultra low binding 96- The wells were cultured for 3 days.
지방 방울 축적을 유도하기 위하여 oleic acid와 plamitic acid를 2:1의 비율로 섞은 지방산 혼합용액을 1 mM이 되도록 간오가노이드에 처리하였다. 동시에 본 발명의 GM-90383과 GM-90803은 DMSO를 이용하여 10 mM이 되도록 제조한후, 순차적으로 희석하고, 최종 농도가 되도록 각각의 웰에 처리하였고 24시간 동안 배양하였다. In order to induce fat droplet accumulation, liver organoids were treated with a fatty acid mixture solution of oleic acid and plamitic acid at a ratio of 2:1 to a concentration of 1 mM. Simultaneously, GM-90383 and GM-90803 of the present invention were prepared to be 10 mM using DMSO, then serially diluted, treated to the final concentration in each well, and cultured for 24 hours.
지방 방울의 염색을 위하여 최종 농도가 10 μM이 되도록 nile-red를 배지에 섞어 주었으며, 분석하고자 하는 영역을 설정하기 위하여 5 μg/mL Hoechst33342으로 핵을 동시에 염색을 수행하였다. 약 15분간 염색을 수행한후, 화합물과 염색 시약들을 제거하기 위하여 HM 배양 배지로 3회 수세한다.For fat droplet staining, nile-red was mixed with the medium to a final concentration of 10 μM, and nuclei were simultaneously stained with 5 μg/mL Hoechst33342 to set the region to be analyzed. After carrying out staining for about 15 minutes, it was washed three times with HM culture medium to remove compounds and staining reagents.
간오가노이드 사진은 Bio-Tek사의 Lion-Heart 장비에서 Gen5 소프트웨어를 사용하여 획득하였다. 세포핵을 염색한 Hoechst33342의 형광은 이미지는 DAPI 필터 (Excitation 377/50 nm, Emmision 447/60 nm)를 사용하여, 지방 방울을 염색한 nile-red의 형광 이미지는 RFP 필터 (Excitation 531/40 nm, Emmision 593/40 nm)를 사용하여 획득하였다. 간오가노이드 내부에 축적된 지방 방울을 정량 분석은 Hoechst33342의 형광 영역을 내의 nile-red 형광 세기를 Gen5 소프트웨어를 사용하여 수행하였다. Liver organoid photographs were acquired using Gen5 software on Bio-Tek's Lion-Heart instrument. The fluorescence image of Hoechst33342 stained cell nuclei was obtained using a DAPI filter (Excitation 377/50 nm, Emmision 447/60 nm), and the fluorescence image of nile-red stained fat droplets was obtained using an RFP filter (Excitation 531/40 nm, Emmison 593/40 nm). Quantitative analysis of fat droplets accumulated inside the liver organoid was performed using Gen5 software by measuring the nile-red fluorescence intensity within the fluorescence region of Hoechst33342.
도 3은 실험예 2에 따라 실시예 1 및 2에서 제조한 화합물을 처리 후 간오가노이드에서의 지방 방울 변화를 형광으로 나타낸 이미지이다. 본 발명의 실시예 1 및 2에서 제조한 화합물의 농도가 증가함에 따라 지방산 혼합액에 의해 증가한 지방 방울의 축적을 감소시킴을 확인할 수 있다.3 is an image showing changes in fat droplets in liver organoids by fluorescence after treatment with the compounds prepared in Examples 1 and 2 according to Experimental Example 2; As the concentration of the compound prepared in Examples 1 and 2 of the present invention increases, it can be confirmed that the accumulation of fat droplets increased by the fatty acid mixture is reduced.
도 4는 상기 도 3의 이미지를 Gen5 소프트웨어를 이용하여 단위 면적당 nile-red 형광의 평균 세기를 분석한 그래프이다. 본 발명의 실시예 2에서 제조한 화합물은 지방산 혼합용액에 의하여 증가한 지방방울의 축적을 감소시켰으며, 특히 5 μM에서 가장 크게 지방 방울의 축적을 감소시켰다. 또한, 실시예 2에서 제조한 화합물은 1과 5 μM의 농도에서 유의적으로 지방 방울의 축적을 감소시키는 것으로 나타났다.FIG. 4 is a graph obtained by analyzing the average intensity of nile-red fluorescence per unit area of the image of FIG. 3 using Gen5 software. The compound prepared in Example 2 of the present invention reduced the accumulation of fat droplets increased by the fatty acid mixture solution, and in particular, reduced the accumulation of fat droplets most significantly at 5 μM. In addition, the compounds prepared in Example 2 were shown to significantly reduce the accumulation of fat droplets at concentrations of 1 and 5 μM.
실험예 3. 제브라피쉬 치어 동물 모델에서의 지방간 억제 효과Experimental Example 3. Fatty liver inhibitory effect in zebrafish fry animal model
타목시펜(tamoxifen)은 에스트로겐 수용체에 결합하는 비스테로이드계 항에스트로겐제로서 영국ICI회사가 개발하여, 1980년대부터 여성호르몬 의존성 암에 내분비요법제로 가장 많이 이용되었다. 특히, 타목시펜을 처리한 제브라피쉬 치어에서는 간세포의 괴사 및 지방증의 소견이 나타나는 것으로 알여져, 본 연구에서는 타목시펜으로 유도된 지방간 모델을 이용하여 실시예 1 및 2에서 제조한 화합물의 지방간 억제 효능을 평가하고자 한다.Tamoxifen (tamoxifen) is a non-steroidal anti-estrogen drug that binds to the estrogen receptor, developed by the British ICI company, and has been used most often as an endocrine therapy for female hormone-dependent cancer since the 1980s. In particular, zebrafish fry treated with tamoxifen are known to show hepatocyte necrosis and steatosis. In this study, the fatty liver inhibitory efficacy of the compounds prepared in Examples 1 and 2 was evaluated using a tamoxifen-induced fatty liver model. want to do
수정 후 5일 시점의 건강한 치어 10 개체를 24-웰 플레이트에 넣고, egg water (60 mg/l sea salt)를 1 ml 넣어준다. 지방간이 조절되지 않은 음성 대조군으로 DMSO를 0.1% 희석하여 치어에 노출시켜주었고, 비알콜성 지방간을 조절하기 위해 타목시펜 (Sigma-Aldrich, T5648)을 5 uM 희석하여 치어에 노출시켰다. 본 발명의 실시예 1 및 2에서 제조한 화합물의 지방간 억제 효능을 알아보기 위해, 5 uM 타목시펜과 각각 혼합하여 1, 2 uM의 실시예 1 및 2에서 제조한 화합물을 치어에 노출시켰다(각 웰 최종 부피는 2 ml). 실시예 1 및 2에서 제조한 화합물을 넣어준 24-웰 플레이트를 호일로 감싸 빛을 차단시켜준 뒤, 28℃ 배양기에서 24시간동안 배양한다. 각 웰의 화합물을 제거하기 위해 egg water로 5분간 3회 수세한다. 지방 특이적 염색을 위해어 LipidGreen2를 5 uM로 희석하여 치어에 30분간 노출시킨 뒤, egg water로 5분간 3회 수세한다. 제브라피쉬 치어의 지방간을 평가하기 위해 Tricaine(5 g/l)으로 치어를 마취한 뒤 3% 메틸 셀룰로즈(Methyl cellulose) 위해 치어를 옮겨 촬영하였다. 촬영 장비는 Lionheart FX Automated Microscope (BioTek)와 Gen5 (BioTek) 소프트웨어를 사용하였다. 사용한 형광 파장은 GFP 필터 세트 (Excitation 469 nm, Emmision 525 nm)를 사용하였다. 이미지상 치어의 간 면적 및 지방 염색 정도는 Gen5 소프트웨어에서 정량하였고, ImageJ 1.50i 소프트웨어(National Institutes of Health, USA)를 이용해 히트맵 이미지로 변환하였다. 형광 현미경 이미지 및 히트맵 변환 이미지상의 점산은 같은 치어의 간 부위를 표시하였으며, 특히, 히트맵 변환 이미지상에서 붉은색 계열로 표시될수록 형광 세기가 강하고, 푸른색 계열로 표시될수록 형광 세기가 약하다는 것을 의미한다. 각 이미지 하단의 그래프는 간부위의 면적 및 형광 세기를 정령한 결과를 나타낸 그래프이다(p 값은 각 실험군 사잉의 t-검정 결과임. *p < 0.05, **p < 0.01, ***p < 0.001).10 healthy fry at 5 days after fertilization are placed in a 24-well plate, and 1 ml of egg water (60 mg/l sea salt) is added. As a negative control in which fatty liver was not controlled, 0.1% DMSO was diluted and exposed to fry, and tamoxifen (Sigma-Aldrich, T5648) was diluted to 5 uM to control non-alcoholic fatty liver and exposed to fry. In order to investigate the fatty liver inhibitory efficacy of the compounds prepared in Examples 1 and 2 of the present invention, 5 uM tamoxifen was mixed with 1 and 2 uM of the compounds prepared in Examples 1 and 2, respectively, and exposed to fry (each well). final volume is 2 ml). The 24-well plate into which the compound prepared in Examples 1 and 2 was put was wrapped with foil to block light, and then incubated in a 28° C. incubator for 24 hours. To remove the compounds in each well, wash with egg water 3 times for 5 minutes. For fat-specific staining, fish LipidGreen2 was diluted to 5 uM and exposed to fry for 30 minutes, followed by washing with egg water three times for 5 minutes. To evaluate the fatty liver of zebrafish fry, after anesthetizing the fry with Tricaine (5 g/l), the fry were transferred to 3% methyl cellulose and photographed. For the imaging equipment, Lionheart FX Automated Microscope (BioTek) and Gen5 (BioTek) software were used. The fluorescence wavelength used was a GFP filter set (Excitation 469 nm, Emmision 525 nm). The liver area and the degree of fat staining of fry on the image were quantified in Gen5 software, and converted to heat map images using ImageJ 1.50i software (National Institutes of Health, USA). Point scattering on the fluorescence microscopy image and the heat map conversion image indicated the liver of the same fry. it means. The graph at the bottom of each image is a graph showing the results of determining the area and fluorescence intensity of the liver (p value is the result of the t-test for each experimental group. *p < 0.05, **p < 0.01, ***p < 0.001).
도 5는 실시예 1에서 제조한 화합물 B의 타목시펜 유도 지방간 모델에서의 지방간 생성 억제 결과이다. 타목시펜 유도 지방군 모델에서는 LipidGreen2 지방 염색 결과 대조군에 비해 형광 세기가 약 41% 증가한다. 그리고 타목시펜 유도 지방간 모델 대비 실시예 1에서 제조한 화합물 1 uM 처리군에서는 약 38%, 2 uM 처리군에서는 약 30% 형광 세기가 감소하는 것으로 보아 지방간의 치료 또는 완화에 유용하게 사용될 수 있음을 알 수 있다.5 is a result of inhibition of fatty liver formation in a tamoxifen-induced fatty liver model by Compound B prepared in Example 1. In the tamoxifen-induced fat group model, as a result of LipidGreen2 fat staining, the fluorescence intensity increased by about 41% compared to the control group. And, compared to the tamoxifen-induced fatty liver model, the compound prepared in Example 1 showed a decrease in fluorescence intensity of about 38% in the 1 uM treatment group and about 30% in the 2 uM treatment group, indicating that it can be usefully used for the treatment or alleviation of fatty liver. can
도 6은 실시예 2에서 제조한 화합물의 타목시펜 유도 지방간 모델에서의 지방간 생성 억제 결과이다. 타목시펜 유도 지방간 모델은 LipidGreen2 지방 염색 결과 대조군에 비해 형광 세기가 약 53% 증가한다. 그리고 타목시펜 유도 지방간 모델 대비 화합물 C 1 uM 처리군에서는 약 54%, 2 uM 처리군에서는 약 43% 형광 세기가 감소하는 것으로 보아 지방간의 치료 또는 완화에 유용하게 사용될 수 있음을 알 수 있다. 6 is a result of inhibition of fatty liver formation in a tamoxifen-induced fatty liver model by the compound prepared in Example 2. As a result of LipidGreen2 fat staining, the fluorescence intensity of the tamoxifen-induced fatty liver model is increased by about 53% compared to the control group. In addition, as compared to the tamoxifen-induced fatty liver model, the fluorescence intensity decreased by about 54% in the compound C 1 uM treatment group and by about 43% in the 2 uM treatment group, indicating that it can be usefully used for the treatment or alleviation of fatty liver.

Claims (14)

  1. 하기 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용 가능한 염:A pyrrolopyrimidine derivative compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
    [화학식 1][Formula 1]
    Figure PCTKR2022011400-appb-img-000010
    Figure PCTKR2022011400-appb-img-000010
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
  2. 제1항에 있어서,According to claim 1,
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 할로젠으로 치환된 페닐인 것을 특징으로 하는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용 가능한 염.In Formula 1, R 1 is N-acetylalanine, or a compound characterized in that a halogen-substituted phenyl, an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
  3. 제1항에 있어서,According to claim 1,
    상기 화학식 1로 표시되는 화합물은 하기 화합물 군으로부터 선택되는 어느 하나인 것을 특징으로 하는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염:The compound represented by Formula 1 is a compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that any one selected from the group of compounds below:
    4-[(4-플루오로벤젠-1-)설피닐]-7H-피롤로[2,3d]피리미딘4-[(4-fluorobenzene-1-)sulfinyl]-7H-pyrrolo[2,3d]pyrimidine
    N-아세틸-S-(7H-피롤로[2,3-d]피리미딘-4-일)-L-시스테인.N-acetyl-S-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-L-cysteine.
  4. 하기 반응식 1에 나타난 바와 같이,As shown in Scheme 1 below,
    화학식 5로 표시되는 화합물로부터 화학식 1로 표시되는 화합물을 제조하는 단계; 를 포함하는 화학식 1로 표시되는 피롤로 피리미딘 유도체 화합물의 제조방법:Preparing a compound represented by Formula 1 from a compound represented by Formula 5; A method for producing a pyrrolopyrimidine derivative compound represented by Formula 1 comprising:
    [반응식 1][Scheme 1]
    Figure PCTKR2022011400-appb-img-000011
    Figure PCTKR2022011400-appb-img-000011
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
  5. 제4항에 있어서,According to claim 4,
    상기 화학식 1의 X가 설피닐인 경우, 하기 반응식 2에 나타난 바와 같이,When X in Formula 1 is sulfinyl, as shown in Scheme 2 below,
    화학식 5로 표시되는 화합물과 화학식 4로 표시되는 화합물을 반응시켜 화학식 3으로 표시되는 화합물을 제조하는 단계(단계 1); 및preparing a compound represented by Chemical Formula 3 by reacting a compound represented by Chemical Formula 5 with a compound represented by Chemical Formula 4 (Step 1); and
    상기 단계 1에서 제조한 화학식 3으로 표시되는 화합물과 옥손(oxone)을 반응시켜 화학식 1-1로 표시되는 화합물을 제조하는 단계(단계 2); 를 포함하는 것을 특징으로 하는, 제조방법:preparing a compound represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone (Step 2); Characterized in that, the manufacturing method comprising:
    [반응식 2] [Scheme 2]
    Figure PCTKR2022011400-appb-img-000012
    Figure PCTKR2022011400-appb-img-000012
    상기 화학식 1에서, R1a는 치환 또는 비치환된 페닐이되,In Formula 1, R 1a is substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환된다.The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen.
  6. 제4항에 있어서,According to claim 4,
    상기 화학식 1의 X가 설파닐인 경우, 하기 반응식 3에 나타난 바와 같이,When X in Formula 1 is sulfanyl, as shown in Scheme 3 below,
    화학식 5로 표시되는 화합물과 화학식 4로 표시되는 화합물을 반응시켜 화학식 3으로 표시되는 화합물을 제조하는 단계(단계 1);preparing a compound represented by Chemical Formula 3 by reacting a compound represented by Chemical Formula 5 with a compound represented by Chemical Formula 4 (Step 1);
    상기 단계 1에서 제조한 화학식 3으로 표시되는 화합물과 옥손(oxone)을 반응시켜 화학식 1-1로 표시되는 화합물을 제조하는 단계(단계 2); 및preparing a compound represented by Chemical Formula 1-1 by reacting the compound represented by Chemical Formula 3 prepared in Step 1 with oxone (Step 2); and
    상기 단계 2에서 제조한 화학식 1-1로 표시되는 화합물과 화학식 2로 표시되는 화합물을 반응시켜 화학식 1-2로 표시되는 화합물을 제조하는 단계(단계 3)를 포함하는 것을 특징으로 하는, 제조방법: characterized in that it comprises a step (step 3) of preparing a compound represented by Chemical Formula 1-2 by reacting the compound represented by Chemical Formula 1-1 prepared in Step 2 with the compound represented by Chemical Formula 2, the manufacturing method :
    [반응식 3][Scheme 3]
    Figure PCTKR2022011400-appb-img-000013
    Figure PCTKR2022011400-appb-img-000013
    상기 화학식 1에서, R1a는 치환 또는 비치환된 페닐이되,In Formula 1, R 1a is substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되며,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    R1b는 N-아세틸알라닌이다.R 1b is N-acetylalanine.
  7. 하기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 치료용 약학적 조성물:A pharmaceutical composition for preventing or treating metabolic liver disease comprising a compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2022011400-appb-img-000014
    Figure PCTKR2022011400-appb-img-000014
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
  8. 제7항에 있어서, According to claim 7,
    상기 화합물은 지방의 축적을 억제하는 것을 특징으로 하는, 약학적 조성물. The pharmaceutical composition, characterized in that the compound inhibits the accumulation of fat.
  9. 제7항에 있어서, According to claim 7,
    상기 대사성 간질환은 알코올성 지방간염 또는 비알코올성 지방간염인 것을 특징으로 하는, 약학적 조성물. The metabolic liver disease is characterized in that alcoholic steatohepatitis or non-alcoholic steatohepatitis, pharmaceutical composition.
  10. 제9항에 있어서, According to claim 9,
    상기 비알코올성 지방간 질환은 단순 지방간 질환, 영양성 지방간 질환, 기아성 지방간 질환, 비만성 지방간 질환, 당뇨병성 지방간 질환 및 지방간염으로 이루어진 군으로부터 선택되는 1 종 이상인 것을 특징으로 하는, 약학적 조성물. The non-alcoholic fatty liver disease is characterized in that at least one selected from the group consisting of simple fatty liver disease, nutritional fatty liver disease, starvation fatty liver disease, obese fatty liver disease, diabetic fatty liver disease and steatohepatitis, pharmaceutical composition.
  11. 하기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 개선용 식품 조성물:A food composition for preventing or improving metabolic liver disease comprising a compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2022011400-appb-img-000015
    Figure PCTKR2022011400-appb-img-000015
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
  12. 제11항에 있어서,According to claim 11,
    상기 식품은 건강기능식품인, 식품 조성물.The food is a health functional food, food composition.
  13. 하기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 대사성 간질환의 예방 또는 개선용 동물 사료 조성물:An animal feed composition for preventing or improving metabolic liver disease comprising a compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
    [화학식 1][Formula 1]
    Figure PCTKR2022011400-appb-img-000016
    Figure PCTKR2022011400-appb-img-000016
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R 1 is N-acetylalanine, or substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
  14. 하기 화학식 1로 표시되는 화합물, 이의 광학 이성질체 또는 이의 약학적으로 허용가능한 염을 대상체에게 치료학적 유효량으로 투여하는 단계를 포함하는 대사성 간질환의 치료방법:A method for treating metabolic liver disease comprising administering to a subject a therapeutically effective amount of a compound represented by Formula 1, an optical isomer thereof, or a pharmaceutically acceptable salt thereof:
    [화학식 1][Formula 1]
    Figure PCTKR2022011400-appb-img-000017
    Figure PCTKR2022011400-appb-img-000017
    상기 화학식 1에서, R1은 N-아세틸알라닌, 또는 치환 또는 비치환된 페닐이되,In Formula 1, R1 is N-acetylalanine or substituted or unsubstituted phenyl,
    상기 치환된 페닐은 히드록시, 시아노, 니트로, 아미노 및 할로젠으로 이루어진 군으로부터 선택되는 1종 이상의 치환기로 치환되고,The substituted phenyl is substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, amino and halogen,
    X는 설피닐 또는 설파닐이다.X is sulfinyl or sulfanyl.
PCT/KR2022/011400 2021-08-02 2022-08-02 Novel pyrrolopyrimidine derivative, method of preparing same, and pharmaceutical composition comprising same as active ingredient for preventing or treating metabolic liver disease WO2023014049A1 (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20140100817A (en) * 2013-02-07 2014-08-18 한국과학기술연구원 7H-Pyrrolo[2,3-d]pyrimidine-4-thiol derivatives using as JAK-3 inhibitors
US20140243312A1 (en) * 2013-02-22 2014-08-28 Pfizer Inc. Pyrrolo[2,3-d]pyrimidine derivatives
WO2019012284A1 (en) * 2017-07-12 2019-01-17 Galapagos Nv Ask1 inhibiting pyrrolopyrimidine and pyrrolopyridine derivatives
KR20200083134A (en) * 2018-12-28 2020-07-08 한국화학연구원 Novel isoquinoline derivative, preparing method thereof, and pharmaceutical composition for preventing or treating autophagy related diseases containing the same as an active ingredient
KR20210044822A (en) * 2018-08-10 2021-04-23 어클라리스 쎄라퓨틱스, 인코포레이티드 Pyrrolopyrimidine ITK inhibitor

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20140100817A (en) * 2013-02-07 2014-08-18 한국과학기술연구원 7H-Pyrrolo[2,3-d]pyrimidine-4-thiol derivatives using as JAK-3 inhibitors
US20140243312A1 (en) * 2013-02-22 2014-08-28 Pfizer Inc. Pyrrolo[2,3-d]pyrimidine derivatives
WO2019012284A1 (en) * 2017-07-12 2019-01-17 Galapagos Nv Ask1 inhibiting pyrrolopyrimidine and pyrrolopyridine derivatives
KR20210044822A (en) * 2018-08-10 2021-04-23 어클라리스 쎄라퓨틱스, 인코포레이티드 Pyrrolopyrimidine ITK inhibitor
KR20200083134A (en) * 2018-12-28 2020-07-08 한국화학연구원 Novel isoquinoline derivative, preparing method thereof, and pharmaceutical composition for preventing or treating autophagy related diseases containing the same as an active ingredient

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