KR20190042393A - Composition for treating inflammatory disease comprising protein kinsase C inhibitor - Google Patents

Abstract

The present invention relates to a composition for alleviating or treating inflammatory diseases containing a protein kinase C inhibitor, or a method for treating inflammatory diseases of an individual comprising a step of administering an effective amount of the composition to the individual. The composition according to an embodiment exhibits excellent effects of inactivating activated macrophages, inhibiting the secretion of inflammatory cytokines, and inhibiting the production of nitric oxide, thereby being able to be used for preventing, alleviating or treating inflammatory diseases.

Description

[0001] The present invention relates to a composition for treating an inflammatory disease comprising a protein kinase C inhibitor,

To a composition for treating or ameliorating an inflammatory disease comprising a protein kinase C inhibitor, or a method of treating an inflammatory disease in a subject comprising administering to said subject an effective amount of said composition.

Inflammatory reactions are collectively referred to as defensive reactions of the body to restore the structure and function of tissues damaged by infection, trauma, and the like. Mobilization of leukocyte cells to the site of inflammation is important for rapid resolution of infection and for repair of tissue damage resulting from various traumas. However, erroneous or persistent inflammatory reactions cause damage and disease of the human body. For example, inflammatory diseases are caused by non-infectious agents such as bacterial or viral infections such as cerebrospinal meningitis, enteritis, dermatitis, uveitis, encephalitis, adult respiratory distress syndrome, trauma, autoimmune diseases and organ transplant rejection.

Septicemia caused by microorganisms in inflammatory diseases is characterized by systemic inflammatory response due to excessive activation of the innate immune system, and thus symptoms or signs such as fever, tachycardia, empty breathing, increase or decrease of leukocyte count are characteristic. When one or more organ failure or hypotension occurs, it is called severe sepsis. It is defined as septic shock when hypotension persists regardless of supply of adequate water or blood pressure. In the United States, sepsis occurs at a rate of 3.9-10.3 per 1,000 people per year, with a mortality rate of 14.7-29.9%. In 2011, the cost of treatment for sepsis patients in the United States has increased to $ 20.3 billion, resulting in increased damage from sepsis.

Conventional sepsis treatment agents are limited to conservative treatments such as supply of fluid, removal of infection source, administration of antibiotics, blood pressure enhancer, etc., in accordance with the patient's condition. There is no licensed therapy that can significantly improve the survival rate by effectively controlling the immune system of sepsis patients. Therefore, there is an urgent need for studies on therapeutic agents for inflammatory diseases including sepsis.

One aspect is to provide a pharmaceutical composition for the prophylaxis or treatment of an inflammatory disease comprising a Protein Kinase C (PKC) inhibitor.

Another aspect is to provide a food composition for improving or alleviating an inflammatory disease comprising a Protein Kinase C (PKC) inhibitor.

Another aspect is to provide a feed composition for improving or alleviating an inflammatory disease comprising protein kinase C (PKC) inhibitor.

Another aspect is to provide a method of preventing or treating sepsis comprising administering to a subject a pharmaceutical composition comprising a Protein Kinase C (PKC) inhibitor or a pharmaceutical composition comprising the same.

One aspect provides a pharmaceutical composition for the prophylaxis or treatment of inflammatory diseases, including protein kinase C (PKC) inhibitors.

As used herein, " Protein Kinase C (PKC) " refers to an enzyme that phosphorylates another protein to regulate the activity, location, and function of the protein to control various intracellular processes.

The PKC inhibitor includes, but is not limited to, substances capable of inhibiting PKC expression or activity, specifically CGP60474 or a pharmaceutically acceptable salt thereof.

As used herein, " CGP60474 " is a PKC inhibitor and can be represented by the following formula. CGP60474 is CGP-60474; 164658-13-3; UNII-16IP6G5MLC; CGP 60474; GNF-Pf-88 < / RTI > The IUPAC designation is based on the name of 3 - [[4- [2- (3-chloroanilo) pyrimidin-4-yl] pyridin- 2- yl] amino] propan- 3-chloroanilino) pyrimidin-4-yl] pyridin-2-yl] amino] propan-1-ol. Although CGP60474 has been reported to be associated with cell cycle regulation, the therapeutic use of sepsis or inflammatory diseases was first identified by the present invention.

[Chemical Formula]

The CGP60474 may comprise a derivative of CGP60474. The term " derivative of CGP60474 " refers to the same activity as CGP60474 and may include, without limitation, substances which have the therapeutic or prophylactic effect of sepsis or inflammatory diseases.

Inflammatory diseases in which the pharmaceutical composition is used are generically referred to as inflammation-predominant diseases, including but not limited to sepsis, rheumatoid arthritis, arteriosclerosis, inflammatory visceral disease, ulcerative colitis, Crohn's disease , Septicemia, meningitis, pancreatitis, peritonitis, vasculitis, lymphocytic choriomeningitis, glomerulonephritis, uveitis, ileitis, liver inflammation, kidney inflammation, rheumatoid arthritis, autoimmune disease, sepsis shock, ischemia, ulcer, diabetes, Alzheimer's Alzheimer's disease, Parkinson's disease, multiple sclerosis, scleroderma, graft rejection, hemorrhagic shock, anaphylactic shock, burn, infection, bacterial infection, viral infection, fungal infection and congenital infection, Hypercholesterolemia, hemodialysis, seizures, cardiovascular transplantation, ischemia / reperfusion disease.

The term " sepsis " refers to a state in which a serious inflammatory reaction occurs in the whole body by infection with microorganisms. The number of breaths increased more than 24 times per minute (ventilation), heart rate more than 90 times per minute (tachycardia), increase in leukocyte count in blood test, or significant decrease in respiratory rate If you have more than one kind of symptoms, it is called systemic inflammatory response syndrome (SIRS). This systemic inflammatory response syndrome is called sepsis when it is caused by microbial infection. It is likely that pathogens enter the bloodstream continuously or intermittently in the infectious lesion of the body, settle in various organs, and cause severe systemic symptoms. Staphylococcus, streptococcus, Escherichia coli, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Pneumococcus, Fungi, and anaerobic bacteria are the causative bacteria.

In the present invention, the term " prevention " means all the actions of inhibiting or delaying the onset of an inflammatory disease upon administration of the composition, and " treatment " it means.

The CGP60474 or a pharmaceutically acceptable salt thereof may be used for the prophylaxis or treatment of sepsis or inflammatory disease by inactivating macrophages, inhibiting the secretion of inflammatory cytokines from macrophages, or inhibiting the production of nitric oxide (NO) May be useful for treatment.

In one embodiment of the invention, the CGP60474 inhibited the secretion of inflammatory cytokines in macrophages activated by treatment with LPS or Poly (I: C). When microorganisms enter the body, LPS and Poly (I: C) among microbial cell wall components bind to toll-like receptors of macrophages to activate macrophages, which is a fundamental cause of sepsis. Thus, septicemia can be fundamentally prevented or treated by inhibiting inflammatory cytokine levels secreted from macrophages activated by LPS, Poly (I: C).

The inflammatory cytokines include interleukin-6 (IL-6), Tumor necrosis factor alpha (TNFa), Interleukin 1b (IL-1b), RANTES, Interleukin-17 IL-17: Interleukin-23: IL-23, CC motif ligand 1: CCL-1, monocyte chemotactic protein 5 : MCP-5), CXC motif chemokine ligand 2: CXCL2, or a combination thereof.

The NO means a radical generated by an inducible nitric oxide synthase (iNOS). Nitric oxide is an inflammatory mediator that is secreted in immune system cells such as macrophages when inflammatory reaction occurs. It has been reported that inhibition of inflammatory reaction by inhibiting the expression of nitric oxide or inhibiting the activity of inducible nitric oxide synthase is reported.

The CGP60474 can be administered in the form of a free form or in the form of a pharmaceutically acceptable salt. The preparation of the medicament can be carried out according to methods known in the art, and commonly known and used adjuvants and further suitable carriers or diluents can be used. The pharmaceutical composition of the present invention may be used in the form of solid, solution, emulsion, dispersant, micelle, liposome, etc., and the resulting composition may be formulated as an active ingredient together with an organic or inorganic carrier or excipient suitable for enteral or parenteral application Include the compounds of the present invention. The active ingredient may be mixed with a generally non-toxic pharmaceutically acceptable carrier, for example, as tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions and any other form suitable for use have. Usable carriers include solid, semisolid or liquid glucose, lactose, gum arabic, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, corn starch, corroid silica, potato starch, urea, Triglycerides, dextrans, of medium chain length, and other carriers suitable for use in the preparation of preparations. Adjuvants, stabilizers, thickeners, coloring agents and perfumes can also be used. The compounds of the present invention are included in the pharmaceutical composition in an amount sufficient to effect a desired effect upon the progression of the disease or the disease state. The pharmaceutical compositions comprising the active ingredient may be in a form suitable for oral use, for example, tablets, rectal tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, A syrup or an elixir. Compositions for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, which may be formulated with suitable excipients such as sucrose, lactose, or saccharin to provide pharmaceutically elegant and mouth- Flavoring agents such as sweeteners, mints, press pods or cherries, coloring agents and preserving agents. In addition, tablets comprising the active ingredient with non-toxic pharmaceutically acceptable excipients can be prepared by known methods. The excipient used is, for example, (1) an inert excipient such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating agents such as corn starch, potato starch or alginic acid and disintegrating agents; (3) binders such as tragacanth gum, corn starch, gelatin or acacia, and (4) lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or coated in a known manner to delay the degradation and absorption in the gastrointestinal tract so that the action may be maintained for a longer period of time. For example, time delay materials such as glyceryl monostearate or glyceryl distearate may be used. They can also be coated to form osmotic therapeutic tablets for controlled release.

In some cases, oral preparations may be used in the form of hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatine capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.

The pharmaceutical composition may be in the form of a sterile injectable suspension. Such suspensions may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents. In addition, sterile injectable preparations can be sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solution, for example as a solution in 1,3-butanediol. Aseptic fixed oils are typically used as a solvent or suspending medium. For this purpose, it is preferred to include synthetic fat delivery agents such as synthetic mono- or diglycerides, fatty acids (including oleic acid), natural vegetable oils such as sesame oil, coconut oil, peanut oil, cottonseed oil, Any non-irritating fixed oil may be used. If necessary, buffering agents, preservatives, antioxidants and the like may be incorporated.

In addition, the compounds useful in the practice of the present invention may be administered in the form of a suppository suitable for rectal administration of the medicament. Such a composition may be prepared by mixing the drug with a suitable non-polar excipient, such as a synthetic glyceride ester of cocoa butter, polyethylene glycol, which is solid at normal temperature but is liquefied and / or dissolved in rectal steel to release the drug.

The pharmaceutical composition may be formulated or used in combination with an anti-inflammatory agent, an antithrombotic agent, a blood coagulation inhibitor, an antibiotic, an antibacterial agent and an anti-allergic agent.

Another aspect provides a method of preventing or treating an inflammatory disease comprising administering to a subject a pharmaceutical composition comprising a Protein Kinase C (PKC) inhibitor.

The above-described PKC inhibitors and inflammatory diseases are as described above.

The term " administering " means introducing the pharmaceutical composition of the present invention to a patient in any appropriate manner, and the administration route can be administered through various routes of oral or parenteral administration, as long as it can reach the target tissue.

The method comprises administering a pharmaceutically effective amount of a PKC inhibitor. It will be apparent to those skilled in the art that the appropriate total daily dose may be determined by the practitioner within the scope of sound medical judgment. In addition, it can be administered once or several times in divided doses. For purposes of the present invention, however, the specific therapeutically effective amount for a particular patient will depend upon the nature and extent of the reaction to be achieved, the particular composition, including whether or not other agents are used, the age, weight, Sex and diet of the patient, the time of administration, the route of administration and the rate of administration of the composition, the duration of the treatment, the drugs used or concurrently used with the specific composition, and similar factors well known in the medical arts. The subject to which the composition is administered includes, but is not limited to, mammals including humans, such as cows, pigs, horses, rabbits, mice, and humans.

Another aspect provides a food composition for improving or alleviating an inflammatory disease comprising a Protein Kinase C (PKC) inhibitor.

The above-described PKC inhibitors and inflammatory diseases are as described above.

The food composition for improving or alleviating the inflammatory disease includes forms such as pills, powders, granules, infusions, tablets, capsules or liquid preparations. Foods to which the composition of the present invention can be added include, for example, , Beverages, gum, tea, vitamin complex, and health supplement foods.

In addition to the composition comprising the composition having the ameliorating or palliative activity of an inflammatory disease comprising the above-mentioned CGP60474 or a pharmaceutically acceptable salt thereof as an essential ingredient that can be contained in the food composition for improving or alleviating inflammatory diseases, There are no limitations, and it may contain various herbal medicine extracts, food-aid additives, natural carbohydrates and the like as additional ingredients such as ordinary foods. Further, as mentioned above, the food-aid additive may be further added, and the food-aid additive includes food-aid additives customary in the art, for example, flavorings, flavors, colorants, fillers, stabilizers and the like .

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .

In addition to the above, the food composition for improving or alleviating an inflammatory disease according to one embodiment of the present invention may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, Chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, it may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

The term " health functional food " refers to a food prepared by processing a specific ingredient as a raw material for the purpose of health assisting or by extracting, concentrating, refining, mixing or the like a specific ingredient contained in a food raw material, It is the food which is designed and processed so that the biomechanical function such as bio-defense, control of the biorhythm, prevention and recovery of disease can be fully demonstrated to the living body.

Another aspect provides a feed composition for improving or alleviating an inflammatory disease comprising protein kinase C (PKC) inhibitor.

The above-described PKC inhibitors and inflammatory diseases are as described above.

The term " feed " means livestock, or any natural or artificial diet, meal, etc., or ingredient of the above formula for eating, ingesting and digesting fish.

The feed may comprise a feed additive or supplementary feed.

The kind of the feed is not particularly limited, and feeds conventionally used in the art can be used. Non-limiting examples of such feeds include vegetable feeds such as cereals, muscle roots, food processing busines logistics, algae, fibers, pharmaceutical buses, oils, fats, pastes or grain by-products; Animal feeds such as proteins, inorganic substances, fats, oils, fats, oils, monocellular proteins, animal plankton or foods. These may be used alone or in combination of two or more.

One aspect provides a pharmaceutical composition for the prophylaxis or treatment of inflammatory diseases, including protein kinase C (PKC) inhibitors.

As used herein, " Protein Kinase C (PKC) " refers to an enzyme that phosphorylates another protein to regulate the activity, location, and function of the protein to control various intracellular processes.

The PKC inhibitor includes, but is not limited to, substances capable of inhibiting PKC expression or activity, specifically CGP60474 or a pharmaceutically acceptable salt thereof.

As used herein, " CGP60474 " is a PKC inhibitor and can be represented by the following formula. CGP60474 is CGP-60474; 164658-13-3; UNII-16IP6G5MLC; CGP 60474; GNF-Pf-88 < / RTI > The IUPAC designation is based on the name of 3 - [[4- [2- (3-chloroanilo) pyrimidin-4-yl] pyridin- 2- yl] amino] propan- 3-chloroanilino) pyrimidin-4-yl] pyridin-2-yl] amino] propan-1-ol. Although CGP60474 has been reported to be associated with cell cycle regulation, the therapeutic use of sepsis or inflammatory diseases was first identified by the present invention.

[Chemical Formula]

The CGP60474 may comprise a derivative of CGP60474. The term " derivative of CGP60474 " refers to the same activity as CGP60474 and may include, without limitation, substances which have the therapeutic or prophylactic effect of sepsis or inflammatory diseases.

Inflammatory diseases in which the pharmaceutical composition is used are generically referred to as inflammation-predominant diseases, including but not limited to sepsis, rheumatoid arthritis, arteriosclerosis, inflammatory visceral disease, ulcerative colitis, Crohn's disease , Septicemia, meningitis, pancreatitis, peritonitis, vasculitis, lymphocytic choriomeningitis, glomerulonephritis, uveitis, ileitis, liver inflammation, kidney inflammation, rheumatoid arthritis, autoimmune disease, sepsis shock, ischemia, ulcer, diabetes, Alzheimer's Alzheimer's disease, Parkinson's disease, multiple sclerosis, scleroderma, graft rejection, hemorrhagic shock, anaphylactic shock, burn, infection, bacterial infection, viral infection, fungal infection and congenital infection, Hypercholesterolemia, hemodialysis, seizures, cardiovascular transplantation, ischemia / reperfusion disease.

The term " sepsis " refers to a state in which a serious inflammatory reaction occurs in the whole body by infection with microorganisms. The number of breaths increased more than 24 times per minute (ventilation), heart rate more than 90 times per minute (tachycardia), increase in leukocyte count in blood test, or significant decrease in respiratory rate If you have more than one kind of symptoms, it is called systemic inflammatory response syndrome (SIRS). This systemic inflammatory response syndrome is called sepsis when it is caused by microbial infection. It is likely that pathogens enter the bloodstream continuously or intermittently in the infectious lesion of the body, settle in various organs, and cause severe systemic symptoms. Staphylococcus, streptococcus, Escherichia coli, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Pneumococcus, Fungi, and anaerobic bacteria are the causative bacteria.

In the present invention, the term " prevention " means all the actions of inhibiting or delaying the onset of an inflammatory disease upon administration of the composition, and " treatment " it means.

The CGP60474 or a pharmaceutically acceptable salt thereof may be used for the prophylaxis or treatment of sepsis or inflammatory disease by inactivating macrophages, inhibiting the secretion of inflammatory cytokines from macrophages, or inhibiting the production of nitric oxide (NO) May be useful for treatment.

In one embodiment of the invention, the CGP60474 inhibited the secretion of inflammatory cytokines in macrophages activated by treatment with LPS or Poly (I: C). When microorganisms enter the body, LPS and Poly (I: C) among microbial cell wall components bind to toll-like receptors of macrophages to activate macrophages, which is a fundamental cause of sepsis. Thus, septicemia can be fundamentally prevented or treated by inhibiting inflammatory cytokine levels secreted from macrophages activated by LPS, Poly (I: C).

The inflammatory cytokines include interleukin-6 (IL-6), Tumor necrosis factor alpha (TNFa), Interleukin 1b (IL-1b), RANTES, Interleukin-17 IL-17: Interleukin-23: IL-23, CC motif ligand 1: CCL-1, monocyte chemotactic protein 5 : MCP-5), CXC motif chemokine ligand 2: CXCL2, or a combination thereof.

The NO means a radical generated by an inducible nitric oxide synthase (iNOS). Nitric oxide is an inflammatory mediator that is secreted in immune system cells such as macrophages when inflammatory reaction occurs. It has been reported that inhibition of inflammatory reaction by inhibiting the expression of nitric oxide or inhibiting the activity of inducible nitric oxide synthase is reported.

The CGP60474 can be administered in the form of a free form or in the form of a pharmaceutically acceptable salt. The preparation of the medicament can be carried out according to methods known in the art, and commonly known and used adjuvants and further suitable carriers or diluents can be used. The pharmaceutical composition of the present invention may be used in the form of solid, solution, emulsion, dispersant, micelle, liposome, etc., and the resulting composition may be formulated as an active ingredient together with an organic or inorganic carrier or excipient suitable for enteral or parenteral application Include the compounds of the present invention. The active ingredient may be mixed with a generally non-toxic pharmaceutically acceptable carrier, for example, as tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions and any other form suitable for use have. Usable carriers include solid, semisolid or liquid glucose, lactose, gum arabic, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, corn starch, corroid silica, potato starch, urea, Triglycerides, dextrans, of medium chain length, and other carriers suitable for use in the preparation of preparations. Adjuvants, stabilizers, thickeners, coloring agents and perfumes can also be used. The compounds of the present invention are included in the pharmaceutical composition in an amount sufficient to effect a desired effect upon the progression of the disease or the disease state. The pharmaceutical compositions comprising the active ingredient may be in a form suitable for oral use, for example, tablets, rectal tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, A syrup or an elixir. Compositions for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, which may be formulated with suitable excipients such as sucrose, lactose, or saccharin to provide pharmaceutically elegant and mouth- Flavoring agents such as sweeteners, mints, press pods or cherries, coloring agents and preserving agents. In addition, tablets comprising the active ingredient with non-toxic pharmaceutically acceptable excipients can be prepared by known methods. The excipient used is, for example, (1) an inert excipient such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating agents such as corn starch, potato starch or alginic acid and disintegrating agents; (3) binders such as tragacanth gum, corn starch, gelatin or acacia, and (4) lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or coated in a known manner to delay the degradation and absorption in the gastrointestinal tract so that the action may be maintained for a longer period of time. For example, time delay materials such as glyceryl monostearate or glyceryl distearate may be used. They can also be coated to form osmotic therapeutic tablets for controlled release.

In some cases, oral preparations may be used in the form of hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatine capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.

The pharmaceutical composition may be in the form of a sterile injectable suspension. Such suspensions may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents. In addition, sterile injectable preparations can be sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solution, for example as a solution in 1,3-butanediol. Aseptic fixed oils are typically used as a solvent or suspending medium. For this purpose, it is preferred to include synthetic fat delivery agents such as synthetic mono- or diglycerides, fatty acids (including oleic acid), natural vegetable oils such as sesame oil, coconut oil, peanut oil, cottonseed oil, Any non-irritating fixed oil may be used. If necessary, buffering agents, preservatives, antioxidants and the like may be incorporated.

In addition, the compounds useful in the practice of the present invention may be administered in the form of a suppository suitable for rectal administration of the medicament. Such a composition may be prepared by mixing the drug with a suitable non-polar excipient, such as a synthetic glyceride ester of cocoa butter, polyethylene glycol, which is solid at normal temperature but is liquefied and / or dissolved in rectal steel to release the drug.

The pharmaceutical composition may be formulated or used in combination with an anti-inflammatory agent, an antithrombotic agent, a blood coagulation inhibitor, an antibiotic, an antibacterial agent and an anti-allergic agent.

Another aspect provides a method of preventing or treating an inflammatory disease comprising administering to a subject a pharmaceutical composition comprising a Protein Kinase C (PKC) inhibitor.

The above-described PKC inhibitors and inflammatory diseases are as described above.

The term " administering " means introducing the pharmaceutical composition of the present invention to a patient in any appropriate manner, and the administration route can be administered through various routes of oral or parenteral administration, as long as it can reach the target tissue.

The method comprises administering a pharmaceutically effective amount of a PKC inhibitor. It will be apparent to those skilled in the art that the appropriate total daily dose may be determined by the practitioner within the scope of sound medical judgment. In addition, it can be administered once or several times in divided doses. For purposes of the present invention, however, the specific therapeutically effective amount for a particular patient will depend upon the nature and extent of the reaction to be achieved, the particular composition, including whether or not other agents are used, the age, weight, Sex and diet of the patient, the time of administration, the route of administration and the rate of administration of the composition, the duration of the treatment, the drugs used or concurrently used with the specific composition, and similar factors well known in the medical arts. The subject to which the composition is administered includes, but is not limited to, mammals including humans, such as cows, pigs, horses, rabbits, mice, and humans.

Another aspect provides a food composition for improving or alleviating an inflammatory disease comprising a Protein Kinase C (PKC) inhibitor.

The above-described PKC inhibitors and inflammatory diseases are as described above.

The food composition for improving or alleviating the inflammatory disease includes forms such as pills, powders, granules, infusions, tablets, capsules or liquid preparations. Foods to which the composition of the present invention can be added include, for example, , Beverages, gum, tea, vitamin complex, and health supplement foods.

In addition to the composition comprising the composition having the ameliorating or palliative activity of an inflammatory disease comprising the above-mentioned CGP60474 or a pharmaceutically acceptable salt thereof as an essential ingredient that can be contained in the food composition for improving or alleviating inflammatory diseases, There are no limitations, and it may contain various herbal medicine extracts, food-aid additives, natural carbohydrates and the like as additional ingredients such as ordinary foods. Further, as mentioned above, the food-aid additive may be further added, and the food-aid additive includes food-aid additives customary in the art, for example, flavorings, flavors, colorants, fillers, stabilizers and the like .

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .

In addition to the above, the food composition for improving or alleviating an inflammatory disease according to one embodiment of the present invention may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, Chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, it may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

The term " health functional food " refers to a food prepared by processing a specific ingredient as a raw material for the purpose of health assisting or by extracting, concentrating, refining, mixing or the like a specific ingredient contained in a food raw material, It is the food which is designed and processed so that the biomechanical function such as bio-defense, control of the biorhythm, prevention and recovery of disease can be fully demonstrated to the living body.

Another aspect provides a feed composition for improving or alleviating an inflammatory disease comprising protein kinase C (PKC) inhibitor.

The above-described PKC inhibitors and inflammatory diseases are as described above.

The term " feed " means livestock, or any natural or artificial diet, meal, etc., or ingredient of the above formula for eating, ingesting and digesting fish.

The feed may comprise a feed additive or supplementary feed.

The kind of the feed is not particularly limited, and feeds conventionally used in the art can be used. Non-limiting examples of such feeds include vegetable feeds such as cereals, muscle roots, food processing busines logistics, algae, fibers, pharmaceutical buses, oils, fats, pastes or grain by-products; Animal feeds such as proteins, inorganic substances, fats, oils, fats, oils, monocellular proteins, animal plankton or foods. These may be used alone or in combination of two or more.

FIG. 1 is a graph illustrating the anti-inflammatory effect of CGP60474 by simultaneously treating LPS and CGP60474 in the macrophage cell line (J774 cell line) and determining IL-6 secretion concentration according to the treatment concentration.
FIG. 2 is a graph showing the concentration of IL-6 secretion according to the treatment concentration of LPS and CGP60474 in the macro cell line (BMDM cell line) simultaneously to confirm the anti-inflammatory effect of CGP60474.
FIG. 3 is a graph showing the anti-inflammatory effect of CGP60474 by pretreating LPS in the macrophage cell line (J774 cell line) for 1 hour and then determining the IL-6 secretion concentration according to the treatment concentration by treating CGP60474.
FIG. 4 is a graph showing the concentration of IL-6 secretion by treatment with CGP60474 after treatment with LPS for 1 hour in the macro cell line (BMDM cell line) to confirm the anti-inflammatory effect of CGP60474.
FIG. 5 is a graph showing the concentration of IL-6 secretion according to the treatment concentration of Poly (I: C) and CGP60474 in the macrophage cell line (J774 cell line) simultaneously to confirm the anti-inflammatory effect of CGP60474.
FIG. 6 is a graph showing the concentration of IL-6 secreted according to the treatment concentration of Poly (I: C) and CGP60474 in the macro cell line (BMDM cell line) simultaneously to confirm the anti-inflammatory effect of CGP60474.
FIG. 7 is a graph showing the anti-inflammatory effect of CGP60474 by pretreating Poly (I: C) for 1 hour with macrophage cell line (J774 cell line) and then examining CGP60474 to determine IL-6 secretion concentration according to treatment concentration.
FIG. 8 is a graph showing the concentration of IL-6 secretion according to the treatment concentration after pretreatment with Poly (I: C) for 1 hour in macrophage cell line (BMDM cell line) and CGP60474 treatment to confirm the anti-inflammatory effect of CGP60474.
9 is a graph showing the NO secretion concentration of CGP60474 treated with macrophage cell line (J774 cell line) in order to confirm the anti-inflammatory effect of CGP60474.
10 is a graph showing the NO secretion level of CGP60474 treated with macrophage cell line (BMDM cell line) in order to confirm the anti-inflammatory effect of CGP60474.
FIG. 11 shows the effect of the LPS alone, LPS, 8nM CGP60474 (LPS + 8nM CGP60474), LPS and 32nM CGP60474 (LPS + 32nM CGP60474) on the macrophage cell line (BMDM cell line) to examine the effect of CGP60474 inhibiting NF- , and p65 were immunostained.
12 is a graph showing the effect of CGP60474 on the inhibition of NF-κB activity of poly (I: C) alone, poly (I: C) and 8nM CGP60474 (Poly (I: C) + 8nM CGP60474) , Poly (I: C) and 32 nM CGP60474 (Poly (I: C) + 32 nM CGP60474) and immunostaining for p50 and p65.
FIG. 13 is a graph showing the results of confirming the survival rate of the mouse over time after induction of sepsis.
14 is a graph showing the concentration of IL-6 in the plasma of the sepsis-induced mouse by ELISA.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these embodiments are intended to illustrate one or more embodiments, and the scope of the present invention is not limited to these embodiments.

Example  One. Of CGP60474  Identify anti-inflammatory effects

In order to confirm the anti-inflammatory effect of CGP60474, the following experiment was conducted.

1.1 LPS  IL-6 induced by concurrent treatment ( Interleukin -6) Confirming the secretion inhibition effect

To induce IL-6 secretion, BMP (bone marrow derived macrophage) cell line and J774 cell line were treated with LPS (Lipopolysaccharide) at 100 ng / ml and CGP60474 at the same time. Changes in IL-6 secretion were confirmed by ELISA. As a control group CGP60474 was used instead of Toll-like receptor-4 inhibitor TAK 272, which is known to have an anti-inflammatory effect. FIG. 1 shows the results of the J774 cell line, and FIG. 2 shows the results of the BMDM cell line.

As shown in FIG. 1 and FIG. 2, it was confirmed that the concentration of IL-6 secretion was similar to that of TAK 272 by CGP60474 co-treated with LPS in both the BMDM cell line and the J774 cell line. In addition, it was confirmed that the concentration of CGP60474 decreased IL-6 secretion in a concentration-dependent manner.

Thus, CGP60474 has been shown to inhibit inflammation through inhibition of IL-6 secretion and macrophage inactivation in superior macrophages.

1.2 LPS  Confirmation of IL-6 secretion inhibition induced by pretreatment

BMDM cell line and J774 cell line were pretreated with 100 ng / ml of LPS. The cell lines in which IL-6 secretion was induced after LPS treatment were treated with CGP60474 and the change of IL-6 secretion was confirmed by ELISA. As a control group CGP60474 was used instead of Toll-like receptor-4 inhibitor TAK 272, which is known to have an anti-inflammatory effect. FIG. 3 shows the results of the J774 cell line, and FIG. 4 shows the results of the BMDM cell line.

As shown in FIG. 3 and FIG. 4, it was confirmed that the concentration of IL-6 secretion induced by pretreatment of LPS was reduced to a level similar to that of TAK 272 by treatment with CGP60474 in both the activated BMDM cell line and the J774 cell line. In addition, it was confirmed that the concentration of CGP60474 decreased IL-6 secretion in a concentration-dependent manner.

Thus, CGP60474 has been shown to inhibit inflammation through inhibition of IL-6 secretion and macrophage inactivation in superior macrophages.

1.3 Poly ( I: C ) Inhibition of IL-6 secretion induced by the simultaneous treatment of IL-6

To induce IL-6 secretion, macrophage BMDM cell line and J774 cell line were treated with LPS at 100 ng / ml and CGP60474 at the same time. Changes in IL-6 secretion were confirmed by ELISA. As a control group (Toll-like receptor-4) inhibitor TAK-242, which is known to have an anti-inflammatory effect, was used instead of CGP60474. FIG. 5 shows the result of the test in the J774 cell line, and FIG. 6 shows the test result in the BMDM cell line.

As shown in FIG. 5 and FIG. 6, it was confirmed that the concentration of IL-6 secretion was reduced to a level similar to that of TAK-242 by CGP60474 co-treated with LPS in both the BMDM cell line and the J774 cell line. In addition, it was confirmed that the concentration of CGP60474 decreased IL-6 secretion in a concentration-dependent manner.

1.4 LPS  Confirmation of IL-6 secretion inhibition induced by pretreatment

BMDM cell line and J774 cell line were pretreated with 100 ng / ml of LPS. The cell lines in which IL-6 secretion was induced after LPS treatment were treated with CGP60474 and the change of IL-6 secretion was confirmed by ELISA. As a control group (Toll-like receptor-4) inhibitor TAK-242, which is known to have an anti-inflammatory effect, was used instead of CGP60474. FIG. 7 shows the results of the J774 cell line, and FIG. 8 shows the results of the BMDM cell line.

As shown in FIG. 7 and FIG. 8, it was confirmed that the concentration of IL-6 secretion induced by pretreatment of LPS was reduced to a level similar to that of TAK-242 by treatment with CGP60474 in both the activated BMDM cell line and the J774 cell line. In addition, it was confirmed that the concentration of CGP60474 decreased IL-6 secretion in a concentration-dependent manner.

Thus, CGP60474 has been shown to inhibit inflammation through inhibition of IL-6 secretion and macrophage inactivation in superior macrophages.

Example  2. Of CGP60474  Identify anti-inflammatory effects

In order to confirm the anti-inflammatory effect of CGP60474, the inhibitory effect of CGP60474 on nitric oxide (NO) production was confirmed as follows. To determine the amount of secreted NO after treatment with LPS in J774 and BMDM cell lines and CGP60474 or TAK-242, the NO concentration in the culture medium was measured by Griess assay.

As a control group (Toll-like receptor-4) inhibitor TAK-242, which is known to have an anti-inflammatory effect, was used instead of CGP60474. FIG. 9 shows the results of the J774 cell line, and FIG. 10 shows the results of the BMDM cell line.

As shown in FIGS. 9 and 10, it was confirmed that the NO secretion level was decreased by CGP60474 in a similar level to that of TAK-242 in both the BMDM cell line and the J774 cell line. In addition, it was confirmed that the NO secretion concentration was decreased in a concentration-dependent manner by CGP60474.

Therefore, CGP60474 has an inhibitory effect on inflammation through inhibition of NO secretion and macrophage inactivation in superior macrophages.

Example  3. Of CGP60474 NF - κB  Identification of the activity inhibition effect

To confirm the inhibitory effect of CGP60474 on NF-kB activity, the following procedure was performed. LPS alone, LPS, 8 nM CGP60474 (LPS + 8 nM CGP60474), LPS and 32 nM CGP60474 (LPS + 32 nM CGP60474) were treated with the mouse macrophage cell line (BMDM cell line) and fixed with 3.5% formaldehyde after 12 hours, The results of immunostaining are shown in Fig.

Similarly, poly (I: C), poly (I: C) and 8nM CGP60474 (Poly (I: C) + 8nM CGP60474), Poly (I: C) and 32nM CGP60474 (I: C) + 32 nM CGP60474), fixed with 3.5% formaldehyde after 12 hours, and subjected to immunostaining for p50 and p65. As a control group, cells treated only with vehicle were used.

As shown in Fig. 11 and Fig. 12, it was confirmed that when CGP60474 was treated together, p50 and p65 were present in cytoplasm more than nuclei, as compared with the case where only LPS or Poly (I: C) was treated.

When NF-κB is activated by inflammation, p50 and p65 migrate to the nucleus. When NF-κB is inactivated, p50 and p65 are present in cytoplasm. Therefore, it was confirmed that CGP60474 inhibits the activation of NF-κB induced by LPS or Poly (I: C) treatment.

Example  4. Of CGP60474  Identification of treatment effect of mouse sepsis

To confirm that CGP60474 is effective in treating sepsis in mice, a sepsis model mouse was prepared by injecting LPS into the abdominal cavity at 10 mg / kg. Immediately after inducing sepsis in mice, mice were treated only with vehicle for 4 days or injected 10 mg / kg CGP60474 once a day into peritoneal cavity and mouse survival rate was confirmed.

FIG. 13 is a graph showing the results of confirming the survival rate of mice after induction of sepsis over time. FIG.

As shown in Fig. 13, it was confirmed that when CGP60474 was injected after 4 days of induction of sepsis, the survival rate was increased about twice as compared to the case where CGP60474 was injected.

14 shows the result of ELISA for the concentration of IL-6 in the plasma after 3 hours from the treatment with vehicle alone or the treatment with CGP60474 in the LPS injected mouse as described above.

As shown in Fig. 14, it was confirmed that the plasma IL-6 concentration in sepsis-induced mice was significantly decreased by treatment with CGP-60474.

Therefore, it was confirmed that CPG06474 can be used for the prevention or treatment of sepsis since the treatment of CPG06474 in mice inducing sepsis leads to an increase in survival rate of mice and inhibition of IL-6 concentration in plasma.

Claims (8)

A pharmaceutical composition for the prophylaxis or treatment of an inflammatory disease comprising a protein kinase C (PKC) inhibitor. The pharmaceutical composition according to claim 1, wherein the PKC inhibitor is CGP60474 or a pharmaceutically acceptable salt thereof. The pharmaceutical composition according to claim 1, which inactivates an activated macrophage. The pharmaceutical composition according to claim 1, which inhibits the secretion of an inflammatory cytokine. The pharmaceutical composition according to claim 1, which inhibits nitric oxide (NO) production. The method according to claim 1, wherein the inflammatory disease is selected from the group consisting of sepsis, rheumatoid arthritis, arteriosclerosis, inflammatory visceral disease, ulcerative colitis, Crohn's disease, encephalitis, meningitis, pancreatitis, peritonitis, vasculitis, A disease selected from the group consisting of uveitis, ileitis, liver inflammation, renal inflammation, rheumatoid arthritis, autoimmune disease, sepsis shock, ischemia, ulcer, diabetes, Alzheimer's disease, Parkinson's disease, multiple sclerosis, Hemorrhagic shock, anaphylactic shock, burns, infections, bacterial infections, viral infections, fungal infections and congenital infections, Rheumatoid arthritis, schistosomiasis, hemodialysis, seizures, cardiovascular transplantation and ischemia / reperfusion Lt; RTI ID = 0.0 > a < / RTI > disorder. A protein composition for improving or alleviating an inflammatory disease comprising a protein kinase C (PKC) inhibitor. A protein composition for improving or alleviating an inflammatory disease comprising protein kinase C (PKC) inhibitor.

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