WO2019194556A1 - Novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound, or pharmaceutical composition for preventing or treating inflammatory bowel diseases and autoimmune diseases comprising same - Google Patents
Novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound, or pharmaceutical composition for preventing or treating inflammatory bowel diseases and autoimmune diseases comprising same Download PDFInfo
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- WO2019194556A1 WO2019194556A1 PCT/KR2019/003908 KR2019003908W WO2019194556A1 WO 2019194556 A1 WO2019194556 A1 WO 2019194556A1 KR 2019003908 W KR2019003908 W KR 2019003908W WO 2019194556 A1 WO2019194556 A1 WO 2019194556A1
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- DSDHOSXCVKCROP-UHFFFAOYSA-N Cc(c(C)c(Nc1nnc(-c(cc2)ccc2OC(F)(F)F)[s]1)nc1C)c1O Chemical compound Cc(c(C)c(Nc1nnc(-c(cc2)ccc2OC(F)(F)F)[s]1)nc1C)c1O DSDHOSXCVKCROP-UHFFFAOYSA-N 0.000 description 1
- KOOBIDSFTUHLHQ-UHFFFAOYSA-N Cc(c(C)c(Nc1nnc(-c(cc2)ccc2OC)[s]1)nc1C)c1O Chemical compound Cc(c(C)c(Nc1nnc(-c(cc2)ccc2OC)[s]1)nc1C)c1O KOOBIDSFTUHLHQ-UHFFFAOYSA-N 0.000 description 1
- YDZGYLCZQJQRCV-UHFFFAOYSA-N Cc(c(C)c(Nc1nnc(-c(cc2)ccc2OCc2ccccc2)[s]1)nc1C)c1O Chemical compound Cc(c(C)c(Nc1nnc(-c(cc2)ccc2OCc2ccccc2)[s]1)nc1C)c1O YDZGYLCZQJQRCV-UHFFFAOYSA-N 0.000 description 1
- XTTJIZYULSLZAU-UHFFFAOYSA-N Cc(c(C)c(Nc1nnc(-c2c[nH]c3ccccc23)[o]1)nc1C)c1O Chemical compound Cc(c(C)c(Nc1nnc(-c2c[nH]c3ccccc23)[o]1)nc1C)c1O XTTJIZYULSLZAU-UHFFFAOYSA-N 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4245—Oxadiazoles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/433—Thidiazoles
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
- A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
Definitions
- the present invention relates to novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compounds, or pharmaceutically acceptable salts thereof.
- the present invention provides a novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound having the effect of preventing or treating inflammatory bowel disease and autoimmune disease, or a pharmaceutically acceptable thereof. It relates to possible salts.
- IBD Inflammatory bowel disease
- Sustained or improper activation of the intestinal immune system plays an important role in the pathophysiology of chronic mucosal inflammation and, in particular, invasion of neutrophils, macrophages, lymphocytes and mast cells results in mucosal destruction and ulceration.
- Infiltrated and activated neutrophils are an important cause of reactive oxygen species and nitric oxide species, which are cytotoxic substances that induce oxidative stress to break down cross-linked proteins, lipids and nucleic acids and lead to epithelial dysfunction and damage do.
- Inflammatory diseases result in the release of various inflammatory cytokines from the intestinal mucosa.
- TNF- ⁇ is elevated in colon lumen and colon epithelial cells of ulcerative colitis patients.
- Recent studies have shown that TNF- ⁇ plays an important role in the pathogenesis of ulcerative colitis.
- Infliximab an anti-TNF- ⁇ antibody, is known to be effective not only for the treatment of rheumatoid arthritis but also for the treatment of Crohn's disease.
- these therapies are costly and in some patients cause side effects such as fluid response or infectious complications.
- IL-6 is one of the cytokine troikas that induce inflammation and is well known as one of the etiologies of inflammatory growth diseases. IL-6 activates JAK after binding to the receptor and then induces phosphorylation of STAT3. Phosphorylated STAT3 acts as a transcription factor to express several types of target genes, including TNF- ⁇ . As such, TNF- ⁇ and IL-6 form a close relationship with each other and are involved in inflammation amplification and tissue damage in inflammatory bowel disease.
- IL-8 is significantly elevated in the mucosa of patients with inflammatory bowel disease and promotes capillary neovascularization. Inflammation in the large intestine increases IL-8 expression, and it has been shown that IL-8 specific antibodies reduce enteritis in animal models of rodents. In this case, the change of intracellular Ca 2+ acts as an important factor of IL-8 induction.
- 5-aminosalicylic acid 5-ASA
- sulfasalazine sulfasalazine
- steroidal immunosuppressants sulfasalazine
- Sulfasalazine is prone to side effects or adverse effects, such as fullness, headache, rash, liver disease, leukopenia, agranulocytosis, male infertility, and the like. It is also unclear whether sulfasalazine has a sufficient relapse inhibitory effect in patients with incisions in the intestine or in patients with remission.
- Steroid immunosuppressants are corticosteroids, and although their short-term effects are recognized, they cannot improve their long-term prognosis. In addition, there are limitations that should be used only in acute cases in terms of side effects such as induced infectious diseases, secondary corticosteroids, peptic ulcers, diabetes, psychiatric disorders, steroidal kidney disease and the like.
- T cells one of the cell groups that play a central role in the immune system, a biological defense system against various pathogens, are generated in the human thymus, and differentiate into T cells having unique characteristics through a series of differentiation processes.
- T cells that have completed differentiation are classified into CD8 T cells and CD4 T cells according to their function.
- CD4 T cells are differentiated in detail according to environmental factors. The most representative ones are Th1, Th2, Th17 and Treg cells.
- Th1 cells are involved in cell mediated immunity
- Th2 cells are involved in humoral immunity
- Th17 cells are involved in infectious and inflammatory diseases
- Treg cells suppress immunity to maintain homeostasis. In the immune system, these cell populations are balanced against each other so that they do not overactivate each other.
- immune diseases can be attributed to the imbalance between these immune cells.
- abnormally increased activity of Th1 and Th17 cells may cause autoimmune diseases
- abnormally increased activity of Th2 cells is known to cause immune diseases due to hypersensitivity.
- Excessive immune suppression of Treg cells can also cause cancer.
- Treg cells have the property of controlling the inflammatory response by inhibiting the function of abnormally activated immune cells. Many experiments have been reported to treat immune diseases through the action of increasing the activity of Treg cells.
- Th1 cells and Th17 cells have been found to be involved in the forefront of the inflammatory response seen in immune diseases, maximizing the signal of the inflammatory response and accelerating disease progression. Therefore, in the case of autoimmune diseases which are not controlled by Treg cells among autoimmune diseases, development of therapeutic agents for autoimmune diseases that target inhibition of Th1 and Th17 cell activities has been highlighted.
- immunosuppressive agents are immunosuppressants that block the signal transduction pathways in T cells. These immunosuppressive agents are toxic, infection, lymphoma, diabetes, tremor, headache, diarrhea, high blood pressure, nausea. Or side effects such as renal failure occurs.
- the inventors have found that the novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound and its pharmaceutically acceptable salts have the effect of preventing or treating excellent inflammatory bowel disease and autoimmune disease. By confirming that the present invention was completed, the present invention was completed.
- the present invention provides novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compounds, or pharmaceutically acceptable salts thereof.
- the present invention also provides a pharmaceutical composition for preventing or treating inflammatory bowel disease or autoimmune disease comprising the compound as an active ingredient.
- the present invention provides the following novel compound of formula (1) or a pharmaceutically acceptable salt thereof.
- Z is O or S
- R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
- alkyl is C 1-30 alkyl
- Alkoxy is C 1-30 alkoxy
- Alkenyl is C 2-30 alkenyl
- Alkynyl is C 2-30 alkynyl
- Cycloalkyl is C 3-30 cycloalkyl
- Aryl is C 5-30 aryl
- Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
- Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
- novel compounds of formula (1) or pharmaceutically acceptable salts thereof of the present invention not only provide excellent inflammatory bowel disease treatment effects, but at the same time provide autoimmune disease treatment effects.
- novel compounds of formula (1) or pharmaceutically acceptable salts thereof of the present invention show superior effects over 5-ASA, the active metabolite of sulfasalazine, a drug currently being used in the treatment of inflammatory bowel disease.
- it is specifically active only in inflammatory cells, and because it is involved only in inflammatory differentiation of T cells, the effect on other cells is less, which has the advantage of less side effects.
- 1A shows the results of cytokine-induced Th1 cell differentiation inhibition by administration of the compound.
- 1B shows the results of cytokine-induced Th17 cell differentiation inhibition by administration of the compound.
- Figure 1C shows the results of the inhibition of differentiation of immunosuppressive cells (FoxP3 +, Treg cells) by administration of the compound.
- 2A to 2C show the results of toxicity experiments on lymphocytes extracted from the spleen after administration of Compound 2-036, tofacitinib and triamcinolone at different concentrations.
- Figure 3A shows the behavior pattern test results of mice in the control group (EAE) and compound 2-036 administration group after antigen injection.
- FIG. 3B and 3C show the result of confirming the number of immune cells in the spleen (FIG. 3B) and the central nervous system (brain and spinal cord) (FIG. 3C) in the control group (EAE) and the compound 2-036 administration group after antigen injection.
- FIG. 3D shows the spinal cords extracted from the control group (EAE) and the compound 2-036-administered group 21 days after antigen injection, cut to 5-10 ⁇ m thickness, and stained with H & E (top panel) and white matter stained with Luxol fast blue. Sectional view (bottom panel).
- FIG. 3E quantifies the number of cells in white matter on the sections stained with H & E of FIG. 3D.
- FIG. 3F quantifies the degree of myelination on the section stained with Luxol fast blue of FIG. 3D.
- Figure 3G is a flow cytometry diagram showing the degree of Th1 and Th17 cell fraction of the cells extracted from the brain and spinal cord of the control group (EAE) and compound 2-036 administration group after antigen injection.
- Figure 3H quantifies the Th1 and Th17 cells distributed in the brain and spinal cord of the control group (EAE) and compound 2-036 administration group after antigen injection.
- FIG. 4A shows the daily weight measurement after the DSS-treated voice control group and the DSS-treated group and the DSS 2-036.
- Figure 4B is a picture of measuring the colon length of the mouse 10 days after the DSS not treated with the negative control group and DSS treatment or DSS 2-036 together and quantified it.
- 4C quantifies the cell number in percent of CD4 T cells and CD8 T cells in the lymph nodes closest to the intestine.
- 4D quantifies the extent of Th1 and Th17 cells in the lymph nodes closest to the intestine.
- the present invention relates to the following novel compounds of formula (1), or pharmaceutically acceptable salts thereof.
- Z is O or S
- R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
- alkyl is C 1-30 alkyl
- Alkoxy is C 1-30 alkoxy
- Alkenyl is C 2-30 alkenyl
- Alkynyl is C 2-30 alkynyl
- Cycloalkyl is C 3-30 cycloalkyl
- Aryl is C 5-30 aryl
- Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
- Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
- the present invention also relates to the following novel compounds of formula (1), or pharmaceutically acceptable salts thereof.
- Z is O or S
- R is hydrogen; Alkyl; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
- alkyl Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl
- alkyl is C 1-18 alkyl
- Alkoxy is C 1-18 alkoxy
- Alkenyl is C 2-18 alkenyl
- Alkynyl is C 2-18 alkynyl
- Cycloalkyl is C 3-18 cycloalkyl
- Aryl is C 5-18 aryl
- Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
- Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
- the compound of the present invention may be a compound represented by Table 1 below, or a pharmaceutically acceptable salt thereof.
- the compound represented by Chemical Formula 2-036 or a pharmaceutically acceptable salt thereof.
- alkyl means a linear or branched hydrocarbon group, which may or may not be substituted with one or more functional groups. Unless otherwise specified, “alkyl” groups preferably contain 1 to 30 carbon atoms. Preferably the “alkyl” group may be, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, heptadecyl.
- Alkoxy refers to —O-alkyl as a functional group to which an alkyl group as defined above is bonded via an oxygen bridge, and may or may not be substituted with one or more functional groups. Preferably it contains 1-30 carbon atoms.
- Alkenyl means a linear, branched or cyclic hydrocarbon group having one or more unsaturated carbon-carbon bonds, which may or may not be substituted with one or more functional groups. Unless otherwise specified, it preferably contains 2 to 30 carbon atoms.
- Alkynyl means a linear or branched hydrocarbon group having one or more carbon-carbon triple bonds, substituted or unsubstituted with one or more functional groups. Unless otherwise specified, it preferably contains 2 to 30 carbon atoms.
- Cycloalkyl means a cyclic or polycyclic hydrocarbon group, which may or may not be substituted with one or more functional groups. Unless otherwise specified, it preferably contains 3 to 30 carbon atoms.
- the "cycloalkyl” group may be, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantane.
- Aryl means a monocyclic or polycyclic aromatic group having 6 to 30 ring carbons, which may or may not be substituted by one or more functional groups.
- the "aryl” group may be, but is not limited to, phenyl, naphthyl.
- Heterocyclic means a monocyclic or polycyclic non-aromatic group in which one or more ring carbons, each substituted or unsubstituted with one or more functional groups, are replaced with heteroatoms selected from B, N, O or S, respectively. Unless otherwise specified, it preferably contains 3 to 30 ring atoms.
- the "heterocyclic” group is piperidinyl, piperazinyl, azetidine, pyrrolidine, 1,3,2-dioxaborolane (1,3,2-dioxaborolane), morpholine, Tetrahydrofuranyl, tetrahydropyranyl, but is not limited thereto.
- Heteroaryl means a monocyclic or polycyclic aromatic group in which one or more ring carbons, each substituted or unsubstituted with one or more functional groups, are replaced with heteroatoms selected from B, N, O or S, respectively. Unless otherwise specified, it preferably contains 3 to 30 ring atoms.
- the “heteroaryl” group is pyrrolyl, triazolyl, tetrazolyl, pyrazolyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, thiadiazolyl, furanyl, Thiophenyl, pyridinyl, pyrazinyl, pyrimidinyl, indole, indazolyl, quinolinyl, isoquinolinyl, benzofuran, benzothiazolyl, benzothiophenyl, pyrazolopyridinyl, but are not limited thereto .
- the fused ring group in which the aryl group is fused to the cycloalkyl group may preferably be tetrahydronaphthyl, but is not limited thereto.
- the fused ring group in which the heteroaryl group is fused to the cycloalkyl group may preferably be tetrahydrocyclopentapyrazolyl, tetrahydroindazolyl or tetrahydrobenzothiophenyl, but is not limited thereto.
- Haloalkyl refers to a branched and straight chain saturated hydrocarbon group in which one or more carbons are replaced with halogen.
- haloalkyl may be, but is not limited to, trifluoromethyl.
- Arylalkoxy can be preferably benzyloxy and aryloxy can be preferably phenoxy, but is not limited thereto.
- the present invention also relates to a pharmaceutical composition having the effect of preventing or treating inflammatory bowel disease, including formula (1) or a pharmaceutically acceptable salt thereof.
- the present invention also relates to a pharmaceutical composition having the effect of preventing or treating an autoimmune disease comprising the formula (1) or a pharmaceutically acceptable salt thereof.
- the present invention also relates to a method for preventing or treating inflammatory bowel disease, comprising administering to a subject a compound of Formula 1 or a pharmaceutically acceptable salt thereof.
- the present invention also relates to a method for preventing or treating an autoimmune disease, comprising administering to a subject a compound of Formula 1 or a pharmaceutically acceptable salt thereof.
- Autoimmune diseases include Behcet's disease, polymyositis, cutaneous myositis, autoimmune cytopenia, autoimmune myocarditis, atopic dermatitis, asthma, primary cirrhosis, dermatitis, fibromyalgia, Goodfight syndrome, autoimmune meningitis, Sjogren's syndrome, systemic Lupus erythematosus, Addison's disease, alopecia areata, ankylosing myelitis, autoimmune hepatitis, autoimmune mumps, insulin dependent diabetes mellitus, dystrophic bullous epidermolysis, epididymitis, glomerulonephritis, Graves' disease, celiac disease, Guillain-Barré syndrome, Hashimoto's disease, Hemolytic anemia, multiple sclerosis, myasthenia gravis, myotrophic lateral sclerosis, vulgaris ulcer, psoriasis, rheumatic fever, rhe
- the inflammatory bowel disease may be selected from the group consisting of ulcerative colitis, Crohn's disease, intestinal Behcet's disease, hemorrhagic rectal ulcer and pouchitis.
- the pharmaceutically acceptable salt is an organic acid selected from the group consisting of oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid and benzoic acid, or an acid moiety formed by an inorganic acid selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid and hydrobromic acid. It may be in the form of a salt, but is not limited thereto.
- compositions of the present invention may be prepared in oral or parenteral preparations.
- Pharmaceutical formulations for oral or parenteral administration can be, for example, in unit dosage form such as tablets, dispersible tablets, coated tablets, effervescent tablets, capsules, suspension powders, suspensions or suppositories, or ampoules. They are produced by known methods, for example by processes such as mixing, granulating, coating or freeze drying.
- compositions for oral administration are prepared by combining the active ingredient with a solid carrier, granulating the mixture obtained if necessary, and processing the mixture or granules, if necessary, after the addition of suitable additives to obtain a tablet or coated tablet core.
- a solid carrier granulating the mixture obtained if necessary
- Suitable carriers may include sugars (eg, lactose, sucrose, mannitol or sorbitol), cellulose, calcium phosphate (eg tricalcium phosphate or calcium hydrogen phosphate), etc. as filler; Binders may include starch (eg, corn, wheat, rice or potato starch), gelatin, tragacanth, methylcellulose, polyvinylpyrrolidone, and the like; Disintegrating agents may include starch, polyvinylpyrrolidone, agar, alginic acid, sodium alginate salts and the like.
- sugars eg, lactose, sucrose, mannitol or sorbitol
- cellulose eg tricalcium phosphate or calcium hydrogen phosphate
- Binders may include starch (eg, corn, wheat, rice or potato starch), gelatin, tragacanth, methylcellulose, polyvinylpyrrolidone, and the like
- Disintegrating agents may include starch, polyviny
- the additive may include salicylic acid, talc, stearic acid, magnesium stearate, calcium stearate, polyethylene glycol, and the like as a lubricant.
- orally administrable pharmaceutical preparations may be hard gelatin capsules and soft sealed capsules of gelatin and plasticizers (eg, glycerol or sorbitol).
- plasticizers eg, glycerol or sorbitol
- aqueous solutions of the active ingredient in water-soluble form for example water-soluble salt form, are mainly suitable.
- the dosage of the active ingredient can be determined depending on various factors such as the activity and duration of action of the active ingredient, the severity or symptoms of the disease to be treated, the method of administration, the species, sex, age, weight of the warm-blooded animal and the individual condition of the warm-blooded animal. have.
- R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
- alkyl is C 1-30 alkyl
- Alkoxy is C 1-30 alkoxy
- Alkenyl is C 2-30 alkenyl
- Alkynyl is C 2-30 alkynyl
- Cycloalkyl is C 3-30 cycloalkyl
- Aryl is C 5-30 aryl
- Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
- Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
- HT-29 human colon cancer-derived epithelial cells and U937 human-derived mononuclear cells were cultured under RPMC 1640 medium containing 10% FBS and 1% penicillin / streptomycin (PS) under 37 ° C. and 5% CO 2 .
- PS penicillin / streptomycin
- HT-29 cells were incubated in a 24 well plate at a concentration of 2 ⁇ 10 5 cells / cm 2 , and the test drug was pretreated for 1 hour in a medium containing only 1% FBS and 1% PS. Thereafter, 10 ⁇ g / mL BCECF-AM was treated and reacted at 37 ° C.
- the inhibitory activity of the test drug (1 ⁇ M) on the adhesion of TNF- ⁇ induced intestinal epithelial cells (HT-29) and mononuclear cells (U937) is shown in Table 4 below.
- 5-ASA positive control
- an active metabolite of sulfasalazine a drug currently used in the treatment of inflammatory bowel disease
- the effect was very small, while 3-073 compound and 2-056 compound showed about 87% inhibition rate, 3-056 compound showed about 81% inhibition rate, and 2-036 compound Showed an inhibition rate of about 79%, showing very good activity.
- Most compounds including compounds 2-041, 2-046, 2-061, 3-046 were found to have superior inhibitory activity compared to the 5-ASA control.
- the 5-ASA positive control
- the 5-ASA showed 1.7% inhibition rate at 1000 ⁇ M concentration for IL-6-induced monocyte-intestinal epithelial cell adhesion, which was 1000 times higher than the test concentration of 1 ⁇ M. While almost no effect was found at the concentration, the 2-056 compound showed about 78% inhibition, the 3-073 compound showed about 77% inhibition, and the 3-056 compound showed about 73% inhibition. It showed excellent activity.
- Compound 2-036 exhibited an inhibition rate of about 69%
- 3-065 exhibited an inhibition rate of about 57%
- 3-047 exhibited an inhibition rate of about 50%, including compounds 2-041, 2-046, and 3-046. It was found that most of the compounds had superior inhibitory activity compared to the 5-ASA control.
- mice were purchased for 7-8 weeks old Sprague Dawley Rat from Orient Bio (Korea) and stabilized with general solid feed for 3 days before being used for the experiment. Feed and water were freely supplied during the experiment, and the temperature in the cage was maintained at 25 ⁇ 1 °C and relative humidity at 50 ⁇ 10%. Lighting control was controlled by a 12-hour light-dark cycle by an automatic light controller.
- the experimental group consisted of 6 animals in each group, and the average weight was 180 ⁇ 10 g by the randomized block design (5 groups (control group, TNBS-only group, TNBS + sulfasalazine 300mg / kg group, TNBS + test drug group)) The experiment was divided by.
- Rats fasted for 24 hours were anesthetized with diethyl ether, and then 5% TNBS (2,4,6-trinitrobenzene sulfonic acid) diluted with 50v / v% ethanol in the colon lumen using a 1 mL syringe connected with polyethylene catheter. After slowly injecting 0.8 mL of the rats, the rats were left upside down for 60 seconds to prevent 5% TNBS from leaking into the anus. The control group was injected with vehicle [50v / v% ethanol] in the same manner as the other groups (Thapa et al., 2008).
- the drug was administered at a fixed time every day for 5 days from the day after TNBS treatment.
- the weight change of each rat was observed from the fasting stage to the TNBS administration and drug administration using a digital mass meter.
- the large intestine of the rat was extracted, and tissues between 5-6 cm from the anus were cut into 1 cm lengths and the tissues were weighed.
- the colon of TNBS-treated rats showed edema and hyperemia compared to the control group, and edema of the appendix and congestion and adhesion of intestinal tissue were observed. .
- the group treated with 300 mg / kg of sulfasalazine, a positive control group also significantly suppressed gross symptoms and adhesions between the other organs and colonic congestion, and sulfasalazine 300 mg in the group administered Compound 2-036 at a dose of 1 mg / kg. Symptoms improved more than the group treated with / kg.
- the weight of the intestine with edema was significantly increased in the TNBS-treated group compared to the vehicle-treated control group, and 300 mg of sulfasalazine, a positive control group.
- the intestinal tissue recovery was 64%.
- Compound 2-036 showed an excellent intestinal weight recovery of 79% even when administered only 1 mg / kg, a dose of 1/300 of sulfasalazine.
- CD4 T cells were isolated from a spleen of B57BL / 6 mice using a magnetic instrument called MACS. In order to stimulate CD4 T cells, CD3 antibody (1mg / mL) was coated on the plate to grow the cells. Then, CD4 T cells were added and other stimulant CD28 antibody (1mg / mL) was added.
- Th1 cells There are several types of inflammatory cells in T cells, the most representative of which are Th1, Th2 or Th17 cells. These inflammatory cells differ in their methods of differentiation.
- Th1 cells 10% FBS and 1% penicillin / streptomycin (PS) were included in RPMI-1640 medium, and the isolated CD4 T cells were incubated under 37 ° C. and 5% CO 2 conditions. When cultured, the cell stimulant CD3 antibody and CD28 antibody were added as before.
- IL-12 (10 ng / mL) was added as a cytokine and grown for 4 days. At this time, compound 2-036 (1 ⁇ M) (treatment group) was included to confirm the inhibitory ability of the drug.
- Th17 cells which are other inflammatory cells
- IL-12 a cytokine
- anti- IFN- ⁇ antibody 5 mg / mL
- anti-IL-4 antibody 5 mg / mL
- IL-2 10 ng / mL
- TGF- ⁇ 10 ng / mL
- the inflammatory cell differentiation inhibitory effect of the compound according to the present invention is shown in Table 7 below.
- Th 1 cell differentiation inhibitory activity (%) Th 17 cell differentiation inhibitory activity (%) 2-036 55 55 2-037 22 24 2-039 27 16 2-041 37 23 2-045 10 30 2-046 26 14 2-047 55 -11 2-048 26 13 2-049 38 22 2-053 18 37 2-056 28 3 2-061 8 9 3-002 23 6 3-010 40 37 3-036 43 8 3-041 33 30 3-046 24 18 3-047 37 16 3-048 27 10 3-049 34 19 3-053 -3 6 3-056 8 -41 3-061 -16 15 3-065 14 20 3-073 One 20 3-086 22 11 3-100 23 23 3-123 25 19
- 2-036 compound showed a inhibition rate of 50% or more in both Th1 and Th17 cells.
- Compound 2-036 was a compound having high inhibition rate and no cytotoxicity under T cell differentiation stimulation conditions.
- Th1 cell differentiation was measured by flow cytometry 3 days after cytokine treatment and is shown in FIG. 1.
- the degree of Th1 differentiation by cytokines was about 30%.
- the degree of cytokine-induced Th1 cell differentiation by the administration of the compound was 13% or less for Compound 2-036, and 8% of tofacitinib and triamcinolone used as positive controls (FIG. 1A).
- Compound 2-036 showed differentiation of 10% or less, tofacitinib, 5%, and triamcinolone about 1% (FIG. 1B).
- tofacitinib and triamcinolone also significantly inhibited the differentiation of immunosuppressive cells (FoxP3 +, Treg cells), whereas Compound 2-036 inhibited the differentiation of Th1 and Th17 cells and did not affect the differentiation of Treg cells, which are immunosuppressive cells. (FIG. 1C).
- Compound 2-036 specifically responds only to inflammatory cells.
- Cells were stimulated under inflammatory cell stimulation conditions for 3 days, and then stained with PI and Annexin V to identify dying cells or dead cells.
- Compound 2-036 shows only inhibitory ability to T cell differentiation and no toxicity to T cell itself, and is evaluated as a drug having a very high specificity for T cell differentiation. Therefore, compound 2-036 is only involved in inflammatory differentiation of T cells, thus showing the possibility of minimizing the effects on other cells and minimizing side effects and maximizing the effect.
- mice B57BL / 6 mice, 8-12 weeks old, were used to feed and water freely during the breeding period, and the temperature of the room was 25 ⁇ 1 °C and relative humidity was 50 ⁇ 10%. Lighting control was controlled by a 12 hour contrast cycle by an automatic light controller.
- the experimental group was divided into 2 groups (control group, drug administration group) by randomized block design with 5 animals in each group.
- EAE experimentalal autoimmune encephalomyelitis
- the control group was intraperitoneally injected with PBS every other day, and the drug administration group was intraperitoneally injected with compound 2-036 every other day in an amount of 3 mg / kg.
- the severity of the disease was divided from 0 to 5.
- the spleen, lymph node (LN), brain and spinal cord (CNS) were extracted from the mouse to calculate the number of immune cells, and the immune cells obtained from each organ to check the distribution of Th1 and Th17 cells.
- LN lymph node
- CNS spinal cord
- the total immune cell number itself was reduced in the drug administration group (FIGS. 3B and 3C), and the inflammatory cells, Th1 and Th17 cells, were also analyzed in the spleen, lymphocyte, It was confirmed that significantly reduced in the brain and spinal cord (Fig. 3G, 3H).
- Compound 2-036 was excellent in inhibiting inflammatory differentiation of T cells and also in inhibiting EAE disease in vivo.
- mice Animals were 8-12 weeks old B57BL / 6 mice, freely supplied with feed and water during the breeding period, and maintained at 25 ⁇ 1 °C and 50 ⁇ 10% relative humidity. Lighting control was controlled by a 12 hour contrast cycle by an automatic light controller.
- the experimental group was divided into three groups (control group, DSS alone administration group, DSS + drug administration group) by randomized block design with 5 animals in each group.
- Enteritis was induced by dissolving DSS in water at 2.5% concentration for 6 days.
- Digital mass meter was used to observe the weight change of each mouse during fasting and DSS and drug administration.
- the colon of the mouse was removed and the length of the tissue was measured.
Abstract
The present invention relates to a novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound, or a pharmaceutically acceptable salt thereof. In particular, the present invention relates to a novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound having an effect of preventing or treating inflammatory bowel diseases and autoimmune diseases, or a pharmaceutically acceptable salt thereof.
Description
본 발명은 신규한 6-헤테로아릴아미노-2,4,5-트라이메틸피리딘-3-올 화합물, 또는 이의 약제학적으로 허용 가능한 염에 관한 것이다. 구체적으로, 본 발명은 염증성 장질환 및 자가면역 질환을 예방 또는 치료하는 효과를 갖는 신규한 6-헤테로아릴아미노-2,4,5-트라이메틸피리딘-3-올 화합물, 또는 이의 약제학적으로 허용 가능한 염에 관한 것이다. The present invention relates to novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compounds, or pharmaceutically acceptable salts thereof. Specifically, the present invention provides a novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound having the effect of preventing or treating inflammatory bowel disease and autoimmune disease, or a pharmaceutically acceptable thereof. It relates to possible salts.
염증성 장질환(Inflammatory bowel disease, IBD)은 임상적으로 유사하면서도, 조직학적 소견과 내시경 및 면역학적 측면에서 서로 다른 궤양성 대장염 및 크론병의 두 가지 질환으로 분류되며 염증세포의 활성화가 중요한 병인인 것으로 알려져 있다.Inflammatory bowel disease (IBD) is clinically similar, but is classified into two types of ulcerative colitis and Crohn's disease in terms of histological findings and endoscopic and immunological aspects. It is known.
장 면역계의 지속적이거나 부적절한 활성화는 만성 점막성 염증의 병태생리에 중요한 역할을 하며, 특히 호중구, 대식세포, 림프구 및 비만세포의 침윤에 의해 결국 점막 파괴 및 궤양을 초래한다. 침윤되고 활성화된 호중구는 활성산소종 및 활성질소종의 중요한 원인이 되며, 이러한 활성종은 세포독성 물질로서 가교 단백질, 지질 및 핵산을 분해하는 산화적 스트레스를 유도하고 상피성 기능장애 및 손상을 초래한다.Sustained or improper activation of the intestinal immune system plays an important role in the pathophysiology of chronic mucosal inflammation and, in particular, invasion of neutrophils, macrophages, lymphocytes and mast cells results in mucosal destruction and ulceration. Infiltrated and activated neutrophils are an important cause of reactive oxygen species and nitric oxide species, which are cytotoxic substances that induce oxidative stress to break down cross-linked proteins, lipids and nucleic acids and lead to epithelial dysfunction and damage do.
염증성 질환이 있으면 장관의 점막에서 다양한 염증성 사이토카인들이 분비된다. TNF-α는 궤양성 대장염 환자의 대장 루멘과 대장 상피세포에서 높게 나타난다. 최근 연구에 의하면, TNF-α는 궤양성 대장염의 병인으로 중요한 역할을 한다고 알려졌다. 항-TNF-α 항체인 인플릭시맵(infliximab)은 류마티스 관절염의 치료뿐 아니라, 기존에 치료되지 않던 크론병의 치료에 효과적이라고 알려졌다. 그러나, 이러한 치료법은 비용이 많이 들고, 일부 환자에게서는 수액 반응 또는 전염성 합병증과 같은 부작용이 야기된다.Inflammatory diseases result in the release of various inflammatory cytokines from the intestinal mucosa. TNF-α is elevated in colon lumen and colon epithelial cells of ulcerative colitis patients. Recent studies have shown that TNF-α plays an important role in the pathogenesis of ulcerative colitis. Infliximab, an anti-TNF-α antibody, is known to be effective not only for the treatment of rheumatoid arthritis but also for the treatment of Crohn's disease. However, these therapies are costly and in some patients cause side effects such as fluid response or infectious complications.
IL-6는 염증을 유도하는 사이토카인 트로이카의 하나이며, 염증성장질환의 병인의 하나로 잘 알려져 있다. IL-6는 수용체와 결합 후 JAK을 활성화시키고 이어서 STAT3의 인산화를 유도한다. 인산화된 STAT3는 전사인자로 작동하여 TNF-α를 비롯한 여러 종류의 타겟유전자를 발현시키게 된다. 이와 같이 TNF-α와 IL-6는 서로 긴밀한 관계를 형성하면서 염증성 장질환에서 염증 증폭 및 조직 손상에 관여하고 있다.IL-6 is one of the cytokine troikas that induce inflammation and is well known as one of the etiologies of inflammatory growth diseases. IL-6 activates JAK after binding to the receptor and then induces phosphorylation of STAT3. Phosphorylated STAT3 acts as a transcription factor to express several types of target genes, including TNF-α. As such, TNF-α and IL-6 form a close relationship with each other and are involved in inflammation amplification and tissue damage in inflammatory bowel disease.
또한, 염증성 장질환을 지닌 환자의 점막에서는 유의성 있게 IL-8이 상승하고, 모세혈관 신생을 촉진하는 것으로 알려져 있다. 대장에서의 염증이 심할수록 IL-8의 발현이 증가하며, 설치류를 이용한 동물실험 모델에서 IL-8 특이적 항체가 장염증을 감소시킨다는 것이 알려졌다. 이때, 세포내 Ca2+의 변화가 IL-8 유도의 중요한 요소로 작용한다.In addition, it is known that IL-8 is significantly elevated in the mucosa of patients with inflammatory bowel disease and promotes capillary neovascularization. Inflammation in the large intestine increases IL-8 expression, and it has been shown that IL-8 specific antibodies reduce enteritis in animal models of rodents. In this case, the change of intracellular Ca 2+ acts as an important factor of IL-8 induction.
현재 염증성 장질환 치료제로는 프로스타글란딘(prostaglandins)의 생성을 차단하는 5-아미노살리실산(5-aminosalicylic acid; 5-ASA) 계통 약물 예를 들어, 설파살라진(sulfasalazine) 등을 이용하거나, 스테로이드류의 면역억제제를 사용하고 있다.Current treatment for inflammatory bowel disease uses 5-aminosalicylic acid (5-ASA) -based drugs that block the production of prostaglandins, such as sulfasalazine, or steroidal immunosuppressants. I'm using.
설파살라진은 복부허실(fullness), 두통, 발진, 간질환, 백혈구 감소증, 무과립구증, 남성 불임 등과 같은 부작용 또는 역효과를 일으키기 쉽다. 또한, 설파살라진이 장의 환부를 절개한 환자 또는 차도가 있는 환자에게 충분한 재발 억제 효과가 있는지는 불분명하다.Sulfasalazine is prone to side effects or adverse effects, such as fullness, headache, rash, liver disease, leukopenia, agranulocytosis, male infertility, and the like. It is also unclear whether sulfasalazine has a sufficient relapse inhibitory effect in patients with incisions in the intestine or in patients with remission.
스테로이드류의 면역억제제는 부신피질 스테로이드로서, 단기적인 효과는 인정받고 있지만, 장기적인 예후를 향상시킬 수는 없다. 또한, 유도된 감염성 질환, 2차 부신피질 부전증, 소화성 궤양, 당뇨병, 정신장애, 스테로이 드성 신장병 등과 같은 부작용의 측면에서 단지 급성인 경우에만 사용되어야 하는 한계가 있다.Steroid immunosuppressants are corticosteroids, and although their short-term effects are recognized, they cannot improve their long-term prognosis. In addition, there are limitations that should be used only in acute cases in terms of side effects such as induced infectious diseases, secondary corticosteroids, peptic ulcers, diabetes, psychiatric disorders, steroidal kidney disease and the like.
즉, 아직까지 염증성 장질환에 대해 신뢰할 만한 경구용 치료요법이 없으므로, 이러한 질환에 대해 효과적이고 저비용의 경구용 치료제의 개발이 요구되고 있다.That is, since there is no reliable oral therapy for inflammatory bowel disease, there is a need for the development of effective and low-cost oral therapy for such diseases.
한편, 각종 병원체에 대한 생체 방어 시스템인 면역계에서 중심적 역할을 담당하는 세포군의 하나인 T 세포는 인체의 흉선에서 생성되며, 일련의 분화 과정을 거치면서 고유의 특성을 지닌 T 세포로 분화한다. 분화를 완료한 T 세포는 그 기능에 따라 크게 CD8 T 세포 및 CD4 T 세포로 구분된다. 이 중 CD4 T 세포는 환경적인 요인에 따라 세부적으로 분화가 되는데, 가장 대표적인 것이 Th1, Th2, Th17 및 Treg 세포이다.Meanwhile, T cells, one of the cell groups that play a central role in the immune system, a biological defense system against various pathogens, are generated in the human thymus, and differentiate into T cells having unique characteristics through a series of differentiation processes. T cells that have completed differentiation are classified into CD8 T cells and CD4 T cells according to their function. Among them, CD4 T cells are differentiated in detail according to environmental factors. The most representative ones are Th1, Th2, Th17 and Treg cells.
Th1 세포는 세포 매개성 면역에 관여하고, Th2 세포는 체액성 면역에 관여하며, Th17 세포는 감염 및 염증 질환에 관여하고, Treg 세포는 면역을 억제하여 항상성을 유지하게 만들어 준다. 면역계에서 이러한 세포 집단들은 서로 과활성화되지 않도록 서로 견제를 통해 면역계의 균형을 유지하고 있다.Th1 cells are involved in cell mediated immunity, Th2 cells are involved in humoral immunity, Th17 cells are involved in infectious and inflammatory diseases, and Treg cells suppress immunity to maintain homeostasis. In the immune system, these cell populations are balanced against each other so that they do not overactivate each other.
면역질환의 대부분은 이러한 면역 세포 간의 불균형에 기인하는 것으로 볼 수 있다. 예를 들어, Th1 세포와 Th17 세포의 활성이 비정상적으로 증가하는 경우 자가면역 질환이 발생할 수 있고, Th2 세포의 활성이 비정상적으로 증가하는 경우 과민반응에 의한 면역질환이 발생하는 것으로 알려져 있다. 또한 Treg 세포의 과도한 면역 억제는 암을 유발할 수 있다.Most of the immune diseases can be attributed to the imbalance between these immune cells. For example, abnormally increased activity of Th1 and Th17 cells may cause autoimmune diseases, and abnormally increased activity of Th2 cells is known to cause immune diseases due to hypersensitivity. Excessive immune suppression of Treg cells can also cause cancer.
최근, Th1 세포와 Th17 세포에 의한 자가면역 질환 연구 결과에 따르면, 이들 세포의 활성을 조절할 수 있는 면역조절 T 세포(Treg)의 기능의 기작이 알려지면서 이를 이용한 면역질환의 치료에 대한 연구가 대두되고 있다. Treg 세포는 비정상적으로 활성화된 면역세포의 기능을 억제하여 염증 반응을 제어하는 특성을 가진다. Treg 세포의 활성을 증가시키는 작용을 통해 면역질환을 치료하는 실험들이 많이 보고되고 있다.Recently, according to a study of autoimmune diseases caused by Th1 and Th17 cells, the mechanism of the function of immunoregulatory T cells (Tregs) that can regulate the activity of these cells is known, and research on the treatment of immune diseases using them has emerged. It is becoming. Treg cells have the property of controlling the inflammatory response by inhibiting the function of abnormally activated immune cells. Many experiments have been reported to treat immune diseases through the action of increasing the activity of Treg cells.
Th1 세포 및 Th17 세포는 면역질환에서 보이는 염증반응의 최전방에서 관여하여 염증 반응의 신호를 최대화 시켜 질병의 진행을 가속화시키는 것이 밝혀지고 있다. 그러므로 자가면역 질환 중 Treg 세포에 의해 제어되지 않는 자가면역 질환의 경우, Th1 및 Th17 세포 활성의 억제를 표적으로 한 자가면역 질환의 치료제 개발이 크게 부각되고 있다.Th1 cells and Th17 cells have been found to be involved in the forefront of the inflammatory response seen in immune diseases, maximizing the signal of the inflammatory response and accelerating disease progression. Therefore, in the case of autoimmune diseases which are not controlled by Treg cells among autoimmune diseases, development of therapeutic agents for autoimmune diseases that target inhibition of Th1 and Th17 cell activities has been highlighted.
현재 사용되고 있는 면역질환 치료제로는 T 세포에서의 신호변환 경로를 차단하는 면역 억제제가 가장 많이 사용되고 있는데, 이러한 면역억제제들은 독성, 감염, 임파종, 당뇨병, 진전(tremor), 두통, 설사, 고혈압, 오심 또는 신기능 장애 등의 부작용이 발생하는 문제점이 있다.Currently used immunosuppressive agents are immunosuppressants that block the signal transduction pathways in T cells. These immunosuppressive agents are toxic, infection, lymphoma, diabetes, tremor, headache, diarrhea, high blood pressure, nausea. Or side effects such as renal failure occurs.
또한, T 세포의 활성화를 억제하는 방법을 통해 면역질환을 치료하는 방법 이외에도, 면역 세포로부터 분비되는 사이토카인의 양을 조절하는 치료법, 및 면역 세포로부터 분비되는 사이토카인을 표적으로 하는 항체를 이용한 치료법이 개발 중에 있다. 그러나 이러한 방법은 실제적으로 임상실험을 거쳐 환자들에게 적용하기까지 많은 시간이 소요 되어야 한다. 또한 항체를 이용하는 방법은 항체 제작 과정에서 너무 많은 비용이 든다는 문제점이 있다.In addition to the treatment of immune diseases through a method of inhibiting the activation of T cells, a therapy for controlling the amount of cytokines secreted from immune cells, and a therapy using antibodies targeting cytokines secreted from immune cells This is under development. However, this method should take a long time to be practically applied to patients after clinical trials. In addition, there is a problem that the method using the antibody is too expensive in the antibody manufacturing process.
따라서 부작용이 없고 저렴하면서도 치료 효과가 우수한 새로운 면역질환 치료제의 개발이 필요한 실정이다. Therefore, there is a need for the development of a new immune disease treatment agent that is inexpensive and has excellent therapeutic effect.
이에, 본 발명자들은 신규한 6-헤테로아릴아미노-2,4,5-트라이메틸피리딘-3-올 화합물 및 이의 약제학적 허용 가능한 염이 우수한 염증성 장질환 및 자가면역 질환을 예방 또는 치료하는 효과를 가짐을 확인함으로써, 본 발명을 완성하였다.Accordingly, the inventors have found that the novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compound and its pharmaceutically acceptable salts have the effect of preventing or treating excellent inflammatory bowel disease and autoimmune disease. By confirming that the present invention was completed, the present invention was completed.
구체적으로, 본 발명은 신규한 6-헤테로아릴아미노-2,4,5-트라이메틸피리딘-3-올 화합물, 또는 이의 약제학적 허용 가능한 염을 제공한다. 또한, 본 발명은 상기 화합물을 유효성분으로 포함하는 염증성 장질환 또는 자가면역 질환의 예방 또는 치료용 약학 조성물을 제공한다. In particular, the present invention provides novel 6-heteroarylamino-2,4,5-trimethylpyridin-3-ol compounds, or pharmaceutically acceptable salts thereof. The present invention also provides a pharmaceutical composition for preventing or treating inflammatory bowel disease or autoimmune disease comprising the compound as an active ingredient.
본 발명은 하기 신규한 화학식 1의 화합물 또는 이의 약제학적으로 허용 가능한 염을 제공한다. The present invention provides the following novel compound of formula (1) or a pharmaceutically acceptable salt thereof.
[화학식 1] [Formula 1]
상기 식에서,Where
Z는 O 또는 S이고,Z is O or S,
R은 수소; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 헤테로아릴알킬; 아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 벤질기로 치환되거나 치환되지 않는 피롤리돈; 알킬벤젠설포네이트; 벤조다이옥솔(benzodioxol); 또는 2,3-디하이드로-1,4-벤조다이옥신(2,3-dihydro-1,4-benzodioxin)이고,R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
상기 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 및 헤테로아릴알킬은The alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl
히드록시; 보로네이트; 할로겐; 할로알킬; 할로알콕시; 할로아릴; 아미노; 알킬아미노; 디알킬아미노; 아실아미노; 아릴카르보닐아미노; 술폰아미노; 니트로; 시아노; 카르보닐; 아실; 아미노카르보닐; 알킬아미노카르보닐; 디알킬아미노카르보닐; 카보히드라지드; 카르복실; 알콕시카르보닐; 아릴술포닐; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 아릴알킬; 디아릴알킬; 아릴알콕시; 알킬 및 할로겐으로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않은 아릴옥시; 알킬로 치환되거나 치환되지 않은 헤테로시클릭; 및 알킬로 치환되거나 치환되지 않은 헤테로아릴로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않으며,Hydroxy; Boronate; halogen; Haloalkyl; Haloalkoxy; Haloaryl; Amino; Alkylamino; Dialkylamino; Acylamino; Arylcarbonylamino; Sulfonamino; Nitro; Cyano; Carbonyl; Acyl; Aminocarbonyl; Alkylaminocarbonyl; Dialkylaminocarbonyl; Carbohydrazide; Carboxyl; Alkoxycarbonyl; Arylsulfonyl; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Arylalkyl; Diarylalkyl; Arylalkoxy; Aryloxy unsubstituted or substituted with one or more groups selected from the group consisting of alkyl and halogen; Heterocyclic, optionally substituted with alkyl; And at least one group selected from the group consisting of heteroaryl unsubstituted or substituted with alkyl,
여기서, 알킬은 C1-30 알킬이고,Wherein alkyl is C 1-30 alkyl,
알콕시는 C1-30 알콕시이고,Alkoxy is C 1-30 alkoxy,
알케닐은 C2-30 알케닐이고,Alkenyl is C 2-30 alkenyl,
알키닐은 C2-30 알키닐이고,Alkynyl is C 2-30 alkynyl,
시클로알킬은 C3-30 시클로알킬이고,Cycloalkyl is C 3-30 cycloalkyl,
아릴은 C5-30 아릴이고,Aryl is C 5-30 aryl,
헤테로시클릭은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로시클릭이고,Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
헤테로아릴은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로아릴이다. Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
본 발명의 신규한 화학식 1의 화합물 또는 이의 약제학적으로 허용 가능한 염은 우수한 염증성 장질환 치료 효과뿐만 아니라, 동시에 자가면역 질환 치료 효과를 제공한다.The novel compounds of formula (1) or pharmaceutically acceptable salts thereof of the present invention not only provide excellent inflammatory bowel disease treatment effects, but at the same time provide autoimmune disease treatment effects.
특히, 본 발명의 신규한 화학식 1의 화합물 또는 이의 약제학적으로 허용 가능한 염은 현재 임상에서 염증성 장질환 치료에 사용되고 있는 약물인 설파살라진의 활성 대사체인 5-ASA에 비해 월등히 우수한 효과를 나타낸다. 또한, 염증 세포에만 특이적으로 활성을 나타내며, T세포의 염증 분화에만 관여하기 때문에 다른 세포에 대한 영향이 적어 부작용이 적은 장점을 갖는다. In particular, the novel compounds of formula (1) or pharmaceutically acceptable salts thereof of the present invention show superior effects over 5-ASA, the active metabolite of sulfasalazine, a drug currently being used in the treatment of inflammatory bowel disease. In addition, it is specifically active only in inflammatory cells, and because it is involved only in inflammatory differentiation of T cells, the effect on other cells is less, which has the advantage of less side effects.
도 1A는 화합물의 투여에 의한 사이토카인-유도성 Th1 세포 분화 억제 실험결과를 나타낸다.1A shows the results of cytokine-induced Th1 cell differentiation inhibition by administration of the compound.
도 1B는 화합물의 투여에 의한 사이토카인-유도성 Th17 세포 분화 억제 실험결과를 나타낸다.1B shows the results of cytokine-induced Th17 cell differentiation inhibition by administration of the compound.
도 1C는 화합물의 투여에 의한 면역억제 세포(FoxP3+, Treg 세포) 분화 억제 실험결과를 나타낸다.Figure 1C shows the results of the inhibition of differentiation of immunosuppressive cells (FoxP3 +, Treg cells) by administration of the compound.
도 2A 내지 2C는, 화합물 2-036, 토파시티닙과 트리암시놀론을 농도별로 투여한 후 비장에서 추출한 임파구세포에 대한 독성 실험결과를 나타낸다.2A to 2C show the results of toxicity experiments on lymphocytes extracted from the spleen after administration of Compound 2-036, tofacitinib and triamcinolone at different concentrations.
도 3A는 항원 주입 후 대조군(EAE) 및 화합물 2-036 투여군에서 마우스의 행동패턴 실험결과를 나타낸다.Figure 3A shows the behavior pattern test results of mice in the control group (EAE) and compound 2-036 administration group after antigen injection.
도 3B와 3C는 항원 주입 후 대조군(EAE) 및 화합물 2-036 투여군에서 비장 (도 3B)과 중추신경(뇌 및 척수)(도 3C)에서의 면역세포 수를 확인한 결과를 나타낸다.3B and 3C show the result of confirming the number of immune cells in the spleen (FIG. 3B) and the central nervous system (brain and spinal cord) (FIG. 3C) in the control group (EAE) and the compound 2-036 administration group after antigen injection.
도 3D는 항원 주입 후 21일째에 대조군(EAE)과 화합물 2-036 투여군에서 척수를 적출하여 5-10 μm 두께로 자른 뒤 H&E로 염색한 단면도(위쪽 패널) 및 Luxol fast blue로 백질을 염색한 단면도(아래 쪽 패널)이다.FIG. 3D shows the spinal cords extracted from the control group (EAE) and the compound 2-036-administered group 21 days after antigen injection, cut to 5-10 μm thickness, and stained with H & E (top panel) and white matter stained with Luxol fast blue. Sectional view (bottom panel).
도 3E는 도 3D의 H&E로 염색한 단면 상의 백질 내 세포 수를 정량한 것이다.FIG. 3E quantifies the number of cells in white matter on the sections stained with H & E of FIG. 3D.
도 3F는 도 3D의 Luxol fast blue로 염색한 단면 상에서 수초화 정도를 정량한 것이다.FIG. 3F quantifies the degree of myelination on the section stained with Luxol fast blue of FIG. 3D.
도 3G는 항원 주입 후 대조군(EAE) 및 화합물 2-036 투여군의 뇌와 척수에서 추출한 세포 중 Th1 및 Th17 세포 분획 정도를 보여주는 유세포분석 결과도이다.Figure 3G is a flow cytometry diagram showing the degree of Th1 and Th17 cell fraction of the cells extracted from the brain and spinal cord of the control group (EAE) and compound 2-036 administration group after antigen injection.
도 3H는 항원 주입 후 대조군(EAE) 및 화합물 2-036 투여군의 뇌와 척수에서 분포하는 Th1 및 Th17 세포 정도를 정량한 것이다.Figure 3H quantifies the Th1 and Th17 cells distributed in the brain and spinal cord of the control group (EAE) and compound 2-036 administration group after antigen injection.
도 4A는 DSS를 처리하지 않은 음성대조군과 DSS 처리한 그룹과 DSS와 2-036을 함께 처리한 후 몸무게를 매일 측정한 것이다.FIG. 4A shows the daily weight measurement after the DSS-treated voice control group and the DSS-treated group and the DSS 2-036.
도 4B는 DSS를 처리하지 않은 음성대조군과 DSS 처리 또는 DSS와 2-036을 함께 처리한 후 10일 뒤 마우스의 대장 길이를 측정한 사진과 이를 정량한 것이다.Figure 4B is a picture of measuring the colon length of the mouse 10 days after the DSS not treated with the negative control group and DSS treatment or DSS 2-036 together and quantified it.
도 4C는 장에 가장 가까운 임파절에서 CD4 T 세포와 CD8 T 세포의 퍼센트로 세포 수를 정량한 것이다.4C quantifies the cell number in percent of CD4 T cells and CD8 T cells in the lymph nodes closest to the intestine.
도 4D는 장에 가장 가까운 임파절에서 Th1 및 Th17 세포의 정도를 정량한 것이다. 4D quantifies the extent of Th1 and Th17 cells in the lymph nodes closest to the intestine.
본 발명은 하기 신규한 화학식 1의 화합물, 또는 이의 약제학적으로 허용 가능한 염에 관한 것이다. The present invention relates to the following novel compounds of formula (1), or pharmaceutically acceptable salts thereof.
[화학식 1] [Formula 1]
상기 식에서,Where
Z는 O 또는 S이고,Z is O or S,
R은 수소; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 헤테로아릴알킬; 아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 벤질기로 치환되거나 치환되지 않는 피롤리돈; 알킬벤젠설포네이트; 벤조다이옥솔(benzodioxol); 또는 2,3-디하이드로-1,4-벤조다이옥신(2,3-dihydro-1,4-benzodioxin)이고,R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
상기 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 및 헤테로아릴알킬은The alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl
히드록시; 보로네이트; 할로겐; 할로알킬; 할로알콕시; 할로아릴; 아미노; 알킬아미노; 디알킬아미노; 아실아미노; 아릴카르보닐아미노; 술폰아미드; 니트로; 시아노; 카르보닐; 아실; 아미노카르보닐; 알킬아미노카르보닐; 디알킬아미노카르보닐; 히드라지드; 카르복실; 알콕시카르보닐; 아릴술포닐; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 아릴알킬; 디아릴알킬; 아릴알콕시; 아세트아미드; 벤즈아미드; 알킬 및 할로겐으로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않은 아릴옥시; 알킬로 치환되거나 치환되지 않은 헤테로시클릭; 및 알킬로 치환되거나 치환되지 않은 헤테로아릴로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않으며,Hydroxy; Boronate; halogen; Haloalkyl; Haloalkoxy; Haloaryl; Amino; Alkylamino; Dialkylamino; Acylamino; Arylcarbonylamino; Sulfonamides; Nitro; Cyano; Carbonyl; Acyl; Aminocarbonyl; Alkylaminocarbonyl; Dialkylaminocarbonyl; Hydrazide; Carboxyl; Alkoxycarbonyl; Arylsulfonyl; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Arylalkyl; Diarylalkyl; Arylalkoxy; Acetamide; Benzamide; Aryloxy unsubstituted or substituted with one or more groups selected from the group consisting of alkyl and halogen; Heterocyclic, optionally substituted with alkyl; And at least one group selected from the group consisting of heteroaryl unsubstituted or substituted with alkyl,
여기서, 알킬은 C1-30 알킬이고,Wherein alkyl is C 1-30 alkyl,
알콕시는 C1-30 알콕시이고,Alkoxy is C 1-30 alkoxy,
알케닐은 C2-30 알케닐이고,Alkenyl is C 2-30 alkenyl,
알키닐은 C2-30 알키닐이고,Alkynyl is C 2-30 alkynyl,
시클로알킬은 C3-30 시클로알킬이고,Cycloalkyl is C 3-30 cycloalkyl,
아릴은 C5-30 아릴이고,Aryl is C 5-30 aryl,
헤테로시클릭은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로시클릭이고,Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
헤테로아릴은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로아릴이다.Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
또한 본 발명은 하기 신규한 화학식 1의 화합물, 또는 이의 약제학적으로 허용 가능한 염에 관한 것이다.The present invention also relates to the following novel compounds of formula (1), or pharmaceutically acceptable salts thereof.
[화학식 1] [Formula 1]
상기 식에서,Where
Z는 O 또는 S이고,Z is O or S,
R은 수소; 알킬; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 헤테로아릴알킬; 아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 벤질기로 치환되거나 치환되지 않는 피롤리돈; 알킬벤젠설포네이트; 벤조다이옥솔(benzodioxol); 또는 2,3-디하이드로-1,4-벤조다이옥신(2,3-dihydro-1,4-benzodioxin)이고,R is hydrogen; Alkyl; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
상기 알킬; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 및 헤테로아릴알킬은The alkyl; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl
히드록시; 보로네이트; 할로겐; 할로알킬; 할로알콕시; 할로아릴; 아미노; 디알킬아미노; 아실아미노; 아릴카르보닐아미노; 술폰아미드; 니트로; 시아노; 히드라지드; 카르복실; 알콕시카르보닐; 아릴술포닐; 알킬; 알콕시; 아릴; 디아릴알킬; 아릴알콕시; 아세트아미드; 벤즈아미드; 알킬 및 할로겐으로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않은 아릴옥시; 알킬로 치환되거나 치환되지 않은 헤테로시클릭; 및 알킬로 치환되거나 치환되지 않은 헤테로아릴로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않으며,Hydroxy; Boronate; halogen; Haloalkyl; Haloalkoxy; Haloaryl; Amino; Dialkylamino; Acylamino; Arylcarbonylamino; Sulfonamides; Nitro; Cyano; Hydrazide; Carboxyl; Alkoxycarbonyl; Arylsulfonyl; Alkyl; Alkoxy; Aryl; Diarylalkyl; Arylalkoxy; Acetamide; Benzamide; Aryloxy unsubstituted or substituted with one or more groups selected from the group consisting of alkyl and halogen; Heterocyclic, optionally substituted with alkyl; And at least one group selected from the group consisting of heteroaryl unsubstituted or substituted with alkyl,
여기서, 알킬은 C1-18 알킬이고,Wherein alkyl is C 1-18 alkyl,
알콕시는 C1-18 알콕시이고,Alkoxy is C 1-18 alkoxy,
알케닐은 C2-18 알케닐이고,Alkenyl is C 2-18 alkenyl,
알키닐은 C2-18 알키닐이고,Alkynyl is C 2-18 alkynyl,
시클로알킬은 C3-18 시클로알킬이고,Cycloalkyl is C 3-18 cycloalkyl,
아릴은 C5-18 아릴이고,Aryl is C 5-18 aryl,
헤테로시클릭은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로시클릭이고,Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
헤테로아릴은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로아릴이다. Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
바람직하게, 본 발명의 화합물은 하기 표 1로 표시되는 화합물, 또는 이의 약제학적으로 허용 가능한 염일 수 있다.Preferably, the compound of the present invention may be a compound represented by Table 1 below, or a pharmaceutically acceptable salt thereof.
더욱 바람직하게는, 상기 표 1에 개시된 화학식 2-036, 2-037, 2-039, 2-041, 2-045, 2-046, 2-047, 2-048, 2-049, 2-053, 2-056, 2-061, 3-002, 3-010, 3-036, 3-041, 3-046, 3-047, 3-048, 3-049, 3-053, 3-056, 3-061, 3-065, 3-073, 3-086, 3-100 또는 3-123로 표시된 화합물, 또는 이의 약제학적으로 허용 가능한 염일 수 있다.More preferably, the chemical formulas 2-036, 2-037, 2-039, 2-041, 2-045, 2-046, 2-047, 2-048, 2-049, and 2-053 disclosed in Table 1 above. , 2-056, 2-061, 3-002, 3-010, 3-036, 3-041, 3-046, 3-047, 3-048, 3-049, 3-053, 3-056, 3 -061, 3-065, 3-073, 3-086, 3-100 or 3-123, or a pharmaceutically acceptable salt thereof.
가장 바람직하게는 상기 화학식 2-036로 표시된 화합물, 또는 이의 약제학적으로 허용 가능한 염이다.Most preferably, the compound represented by Chemical Formula 2-036, or a pharmaceutically acceptable salt thereof.
본 발명에서, "알킬"은 하나 이상의 작용기로 치환되거나 치환되지 않는, 선형 또는 분지형의 탄화수소기를 의미한다. 달리 특정되지 않는 한, "알킬" 기는 바람직하게는 1 내지 30개의 탄소수를 함유한다. 바람직하게는 “알킬”기는 메틸, 에틸, 프로필, 이소프로필, 부틸, 헵타데실일 수 있으나, 이에 제한되지 않는다.In the present invention, "alkyl" means a linear or branched hydrocarbon group, which may or may not be substituted with one or more functional groups. Unless otherwise specified, "alkyl" groups preferably contain 1 to 30 carbon atoms. Preferably the "alkyl" group may be, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, heptadecyl.
"알콕시"는 상기 정의된 알킬기가 산소 가교(oxygen bridge)를 통해 결합되는 작용기로 -O-알킬을 나타내며, 하나 이상의 작용기로 치환되거나 치환되지 않는다. 바람직하게는 1 내지 30개의 탄소수를 함유한다.“Alkoxy” refers to —O-alkyl as a functional group to which an alkyl group as defined above is bonded via an oxygen bridge, and may or may not be substituted with one or more functional groups. Preferably it contains 1-30 carbon atoms.
"알케닐"은 하나 이상의 작용기로 치환되거나 치환되지 않는, 하나 이상의 불포화 탄소-탄소 결합을 갖는 선형, 분지형 또는 고리형의 탄화수소기를 의미한다. 달리 특정되지 않는 한, 바람직하게는 2 내지 30개의 탄소수를 함유한다."Alkenyl" means a linear, branched or cyclic hydrocarbon group having one or more unsaturated carbon-carbon bonds, which may or may not be substituted with one or more functional groups. Unless otherwise specified, it preferably contains 2 to 30 carbon atoms.
"알키닐"은 하나 이상의 작용기로 치환되거나 치환되지 않는, 하나 이상의 탄소-탄소 삼중 결합을 갖는 선형 또는 분지형의 탄화수소기를 의미한다. 달리 특정되지 않는 한, 바람직하게는 2 내지 30개의 탄소수를 함유한다."Alkynyl" means a linear or branched hydrocarbon group having one or more carbon-carbon triple bonds, substituted or unsubstituted with one or more functional groups. Unless otherwise specified, it preferably contains 2 to 30 carbon atoms.
“시클로알킬”은 하나 이상의 작용기로 치환되거나 치환되지 않는, 고리형 또는 다중고리형 탄화수소기를 의미한다. 달리 특정되지 않는 한, 바람직하게는 3 내지 30개의 탄소수를 함유한다. 바람직하게는 “시클로알킬”기는 시클로프로필, 시클로부틸, 시클로펜틸, 시클로헥실, 아다만탄일 수 있으나, 이에 제한되는 것은 아니다."Cycloalkyl" means a cyclic or polycyclic hydrocarbon group, which may or may not be substituted with one or more functional groups. Unless otherwise specified, it preferably contains 3 to 30 carbon atoms. Preferably, the "cycloalkyl" group may be, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantane.
“아릴”은 하나 이상의 작용기로 치환되거나 치환되지 않는, 6 내지 30개의 고리 탄소를 갖는 단환 또는 다환의 방향족기를 의미한다. 바람직하게는 “아릴”기는 페닐, 나프틸일 수 있으나, 이에 제한되는 것은 아니다."Aryl" means a monocyclic or polycyclic aromatic group having 6 to 30 ring carbons, which may or may not be substituted by one or more functional groups. Preferably, the "aryl" group may be, but is not limited to, phenyl, naphthyl.
“헤테로시클릭”은 하나 이상의 작용기로 치환되거나 치환되지 않는, 하나 이상의 고리 탄소가 각각 B, N, O 또는 S로부터 선택된 헤테로원자로 대체되는 단환 또는 다환의 비-방향족기를 의미한다. 달리 특정되지 않는 한, 바람직하게는 3 내지 30개의 고리 원자를 함유한다. 바람직하게는 “헤테로시클릭”기는 피페리디닐, 피페라지닐, 아제티딘, 피롤리딘, 1,3,2-디옥사보롤란(1,3,2-dioxaborolane), 모르폴린(morpholine), 테트라히드로퓨라닐, 테트라히드로피라닐일 수 있으나, 이에 제한되는 것은 아니다."Heterocyclic" means a monocyclic or polycyclic non-aromatic group in which one or more ring carbons, each substituted or unsubstituted with one or more functional groups, are replaced with heteroatoms selected from B, N, O or S, respectively. Unless otherwise specified, it preferably contains 3 to 30 ring atoms. Preferably, the "heterocyclic" group is piperidinyl, piperazinyl, azetidine, pyrrolidine, 1,3,2-dioxaborolane (1,3,2-dioxaborolane), morpholine, Tetrahydrofuranyl, tetrahydropyranyl, but is not limited thereto.
“헤테로아릴”은 하나 이상의 작용기로 치환되거나 치환되지 않는, 하나 이상의 고리 탄소가 각각 B, N, O 또는 S로부터 선택된 헤테로원자로 대체되는 단환 또는 다환의 방향족기를 의미한다. 달리 특정되지 않는 한, 바람직하게는 3 내지 30개의 고리 원자를 함유한다. 바람직하게는 “헤테로아릴”기는 피롤릴, 트리아졸릴, 테트라졸릴, 피라졸일, 옥사졸릴(oxazolyl), 옥사디아졸릴 (oxadiazolyl), 이속사졸릴(isoxazolyl), 티아졸릴, 티아디아졸릴, 퓨라닐, 티오페닐, 피리디닐, 피라지닐, 피리미디닐, 인돌, 인다졸릴, 퀴놀리닐, 이소퀴놀리닐, 벤조퓨란, 벤조티아졸릴, 벤조티오페닐, 피라졸로피리디닐일 수 있으나, 이에 제한되지 않는다."Heteroaryl" means a monocyclic or polycyclic aromatic group in which one or more ring carbons, each substituted or unsubstituted with one or more functional groups, are replaced with heteroatoms selected from B, N, O or S, respectively. Unless otherwise specified, it preferably contains 3 to 30 ring atoms. Preferably, the “heteroaryl” group is pyrrolyl, triazolyl, tetrazolyl, pyrazolyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, thiadiazolyl, furanyl, Thiophenyl, pyridinyl, pyrazinyl, pyrimidinyl, indole, indazolyl, quinolinyl, isoquinolinyl, benzofuran, benzothiazolyl, benzothiophenyl, pyrazolopyridinyl, but are not limited thereto .
아릴기가 시클로알킬기에 융합되어 있는 융합고리기는 바람직하게는 테트라히드로나프틸일 수 있으나, 이에 제한되지 않는다.The fused ring group in which the aryl group is fused to the cycloalkyl group may preferably be tetrahydronaphthyl, but is not limited thereto.
헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기는 바람직하게는 테트라히드로시클로펜타피라졸릴, 테트라히드로인다졸릴 또는 테트라히드로벤조티오페닐일 수 있으나, 이에 제한되지 않는다.The fused ring group in which the heteroaryl group is fused to the cycloalkyl group may preferably be tetrahydrocyclopentapyrazolyl, tetrahydroindazolyl or tetrahydrobenzothiophenyl, but is not limited thereto.
"할로알킬"은 하나 이상의 탄소가 할로겐으로 치환된 분지쇄 및 직쇄의 포화된 탄화수소기를 의미한다. 바람직하게는, “할로알킬”은 트리플루오로메틸일 수 있으나, 이에 제한되지 않는다."Haloalkyl" refers to a branched and straight chain saturated hydrocarbon group in which one or more carbons are replaced with halogen. Preferably, “haloalkyl” may be, but is not limited to, trifluoromethyl.
아릴알콕시는 바람직하게는 벤질옥시일 수 있고, 아릴옥시는 바람직하게는 페녹시일 수 있으나, 이에 제한되지 않는다. Arylalkoxy can be preferably benzyloxy and aryloxy can be preferably phenoxy, but is not limited thereto.
또한, 본 발명은 화학식 1 또는 이의 약제학적으로 허용 가능한 염을 포함하는 염증성 장질환을 예방 또는 치료하는 효과를 갖는 약학적 조성물에 관한 것이다.The present invention also relates to a pharmaceutical composition having the effect of preventing or treating inflammatory bowel disease, including formula (1) or a pharmaceutically acceptable salt thereof.
또한, 본 발명은 화학식 1 또는 이의 약제학적으로 허용 가능한 염을 포함하는 자가면역 질환을 예방 또는 치료하는 효과를 갖는 약학적 조성물에 관한 것이다.The present invention also relates to a pharmaceutical composition having the effect of preventing or treating an autoimmune disease comprising the formula (1) or a pharmaceutically acceptable salt thereof.
또한, 본 발명은 화학식 1의 화합물 또는 이의 약제학적으로 허용 가능한 염을 대상체에 투여하는 것을 포함하는, 염증성 장질환의 예방 또는 치료방법에 관한 것이다.The present invention also relates to a method for preventing or treating inflammatory bowel disease, comprising administering to a subject a compound of Formula 1 or a pharmaceutically acceptable salt thereof.
또한, 본 발명은 화학식 1의 화합물 또는 이의 약제학적으로 허용 가능한 염을 대상체에 투여하는 것을 포함하는, 자가면역 질환의 예방 또는 치료방법에 관한 것이다.The present invention also relates to a method for preventing or treating an autoimmune disease, comprising administering to a subject a compound of Formula 1 or a pharmaceutically acceptable salt thereof.
자가면역 질환은 베체트병, 다발성 근육염, 피부 근육염, 자가면역 혈구감소증, 자가면역 심근염, 아토피피부염, 천식, 일차성간경변, 피부근염, 섬유근육통, 굿파이처 증후군, 자가면역 뇌수막염, 쇼그렌 증후군, 전신 홍반성 루프스, 애디슨병, 원형 탈모증, 강직성 척수염, 자가면역성 간염, 자가면역성 이하선염, 인슐린 의존성 당뇨병, 이영양성 수포성 표피박리증, 부고환염, 사구체 신염, 그레이브스병, 셀리악병, 길랑바레 증후군, 하시모토병, 용혈성 빈혈, 다발성 경화증, 중증 근무력증, 근위축성측색경화증, 심상천포창, 건선, 류마티스열, 류마티스 관절염, 유육종증, 피부 경화증, 척추관절증, 갑상선염, 혈관염, 백반증, 점액수종, 악성빈혈, 항인지질증후군, 고형장기 이식 후기 및 만성 거부증, 또는 이식편대숙주질환일 수 있다.Autoimmune diseases include Behcet's disease, polymyositis, cutaneous myositis, autoimmune cytopenia, autoimmune myocarditis, atopic dermatitis, asthma, primary cirrhosis, dermatitis, fibromyalgia, Goodfight syndrome, autoimmune meningitis, Sjogren's syndrome, systemic Lupus erythematosus, Addison's disease, alopecia areata, ankylosing myelitis, autoimmune hepatitis, autoimmune mumps, insulin dependent diabetes mellitus, dystrophic bullous epidermolysis, epididymitis, glomerulonephritis, Graves' disease, celiac disease, Guillain-Barré syndrome, Hashimoto's disease, Hemolytic anemia, multiple sclerosis, myasthenia gravis, myotrophic lateral sclerosis, vulgaris ulcer, psoriasis, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, spondyloarthropathy, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, antiphospholipid syndrome, solid Organ transplant late and chronic rejection, or graft-versus-host disease.
상기 염증성 장질환은 궤양성 대장염, 크론병(Crohn's disease), 장관형 베체트병, 출혈성 직장 궤양 및 회장낭 염(pouchitis)으로 이루어진 군에서 선택될 수 있다. The inflammatory bowel disease may be selected from the group consisting of ulcerative colitis, Crohn's disease, intestinal Behcet's disease, hemorrhagic rectal ulcer and pouchitis.
상기 약제학적 허용 가능한 염은 옥살산, 말레산, 푸마르산, 말산, 타르타르산, 시트르산 및 벤조산으로 이루어 진 군에서 선택된 유기산이거나, 또는 염산, 황산, 인산 및 브롬화수소산으로 이루어진 군에서 선택된 무기산에 의해 형성되는 산부가염의 형태일 수 있으나, 이에 제한되지 않는다. The pharmaceutically acceptable salt is an organic acid selected from the group consisting of oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid and benzoic acid, or an acid moiety formed by an inorganic acid selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid and hydrobromic acid. It may be in the form of a salt, but is not limited thereto.
또한, 본 발명의 약학적 조성물은 경구 또는 비경구용 제제로 제조될 수 있다. 경구 또는 비경구 투여를 위한 약제학적 제제는, 예를 들면, 정제, 분산성 정제, 코팅정, 발포성 정제, 캡슐, 현탁성 분말제, 현탁액제 또는 좌약, 또는 앰풀과 같은 단위 제형일 수 있다. 이들은 공지된 방법으로, 예를 들어 혼합, 과립화, 코팅 또는 동결 건조 등의 공정에 의해 제조된다.In addition, the pharmaceutical compositions of the present invention may be prepared in oral or parenteral preparations. Pharmaceutical formulations for oral or parenteral administration can be, for example, in unit dosage form such as tablets, dispersible tablets, coated tablets, effervescent tablets, capsules, suspension powders, suspensions or suppositories, or ampoules. They are produced by known methods, for example by processes such as mixing, granulating, coating or freeze drying.
경구 투여를 위한 약제학적 제제는 활성 성분을 고형 담체와 조합하고, 필요하다면 수득한 혼합물을 과립화하고, 혼합물 또는 과립을, 필요하다면 적당한 첨가제의 첨가 후 가공하여 정제 또는 코팅정 코어를 수득함으로써 제조될 수 있다.Pharmaceutical preparations for oral administration are prepared by combining the active ingredient with a solid carrier, granulating the mixture obtained if necessary, and processing the mixture or granules, if necessary, after the addition of suitable additives to obtain a tablet or coated tablet core. Can be.
상기 적당한 담체는, 충전제로 당 (예를 들어, 락토오스, 수크로스, 만니톨 또는 솔비톨), 셀룰로오스, 인산칼슘 (예를 들어, 인산삼칼슘 또는 인산수소칼슘) 등을 포함할 수 있으며; 결합제로 전분 (예를 들어, 옥수수, 밀, 쌀 또는 감자 전분), 젤라틴, 트라가칸트, 메틸셀룰로오스, 폴리비닐피롤리돈 등을 포함할 수 있으며; 붕해제로 전분, 폴리비닐피롤리돈, 한천, 알긴산, 또는 알긴산나트륨염 등을 포함할 수 있다.Suitable carriers may include sugars (eg, lactose, sucrose, mannitol or sorbitol), cellulose, calcium phosphate (eg tricalcium phosphate or calcium hydrogen phosphate), etc. as filler; Binders may include starch (eg, corn, wheat, rice or potato starch), gelatin, tragacanth, methylcellulose, polyvinylpyrrolidone, and the like; Disintegrating agents may include starch, polyvinylpyrrolidone, agar, alginic acid, sodium alginate salts and the like.
상기 첨가제는 윤활제로 살리실산, 활석, 스테아르산, 스테아르산 마그네슘, 스테아르산 칼슘, 폴리에틸렌 글리콜 등을 포함할 수 있다.The additive may include salicylic acid, talc, stearic acid, magnesium stearate, calcium stearate, polyethylene glycol, and the like as a lubricant.
또한, 경구 투여 가능한 약제학적 제제는 경질의 젤라틴 캡슐, 및 젤라틴 및 가소제(예를 들어, 글리세롤 또는 솔비톨)의 연질 밀봉 캡슐일 수 있다.In addition, orally administrable pharmaceutical preparations may be hard gelatin capsules and soft sealed capsules of gelatin and plasticizers (eg, glycerol or sorbitol).
비경구적 투여의 경우는, 수용성 형태, 예를 들면 수용성 염 형태의 활성 성분의 수용액이 주로 적당하다.For parenteral administration, aqueous solutions of the active ingredient in water-soluble form, for example water-soluble salt form, are mainly suitable.
활성 성분의 복용량은, 활성 성분의 활성 및 작용 지속 기간, 치료할 질병의 심각성 또는 그의 증상, 투여 방법, 온혈 동물의 종, 성별, 나이, 체중 및 온혈 동물의 개별적인 상태와 같은 각종 요인에 따라 결정할 수 있다. The dosage of the active ingredient can be determined depending on various factors such as the activity and duration of action of the active ingredient, the severity or symptoms of the disease to be treated, the method of administration, the species, sex, age, weight of the warm-blooded animal and the individual condition of the warm-blooded animal. have.
본 발명의 화학식 1로 표시되는 화합물은 하기 반응식 1 내지 3과 같은 제조방법으로 제조하였다.Compound represented by Formula 1 of the present invention was prepared by the same method as in Schemes 1 to 3.
[반응식 1] Scheme 1
상기 VIII 및 IX에서,In VIII and IX above,
R은 수소; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 헤테로아릴알킬; 아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 벤질기로 치환되거나 치환되지 않는 피롤리돈; 알킬벤젠설포네이트; 벤조다이옥솔(benzodioxol); 또는 2,3-디하이드로-1,4-벤조다이옥신(2,3-dihydro-1,4-benzodioxin)이고,R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),
상기 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 및 헤테로아릴알킬은The alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl
히드록시; 보로네이트; 할로겐; 할로알킬; 할로알콕시; 할로아릴; 아미노; 알킬아미노; 디알킬아미노; 아실아미노; 아릴카르보닐아미노; 술폰아미드; 니트로; 시아노; 카르보닐; 아실; 아미노카르보닐; 알킬아미노카르보닐; 디알킬아미노카르보닐; 히드라지드; 카르복실; 알콕시카르보닐; 아릴술포닐; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 아릴알킬; 디아릴알킬; 아릴알콕시; 아세트아미드; 벤즈아미드; 알킬 및 할로겐으로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않은 아릴옥시; 알킬로 치환되거나 치환되지 않은 헤테로시클릭; 및 알킬로 치환되거나 치환되지 않은 헤테로아릴로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않으며,Hydroxy; Boronate; halogen; Haloalkyl; Haloalkoxy; Haloaryl; Amino; Alkylamino; Dialkylamino; Acylamino; Arylcarbonylamino; Sulfonamides; Nitro; Cyano; Carbonyl; Acyl; Aminocarbonyl; Alkylaminocarbonyl; Dialkylaminocarbonyl; Hydrazide; Carboxyl; Alkoxycarbonyl; Arylsulfonyl; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Arylalkyl; Diarylalkyl; Arylalkoxy; Acetamide; Benzamide; Aryloxy unsubstituted or substituted with one or more groups selected from the group consisting of alkyl and halogen; Heterocyclic, optionally substituted with alkyl; And at least one group selected from the group consisting of heteroaryl unsubstituted or substituted with alkyl,
여기서, 알킬은 C1-30 알킬이고,Wherein alkyl is C 1-30 alkyl,
알콕시는 C1-30 알콕시이고,Alkoxy is C 1-30 alkoxy,
알케닐은 C2-30 알케닐이고,Alkenyl is C 2-30 alkenyl,
알키닐은 C2-30 알키닐이고,Alkynyl is C 2-30 alkynyl,
시클로알킬은 C3-30 시클로알킬이고,Cycloalkyl is C 3-30 cycloalkyl,
아릴은 C5-30 아릴이고,Aryl is C 5-30 aryl,
헤테로시클릭은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로시클릭이고,Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,
헤테로아릴은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로아릴이다. Heteroaryl is a heteroaryl having 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S.
[반응식 2] Scheme 2
상기 반응식 1에서 수득한 화학식 IX의 화합물로부터 하기 화학식 2의 화합물을 하기 반응식 2에 따라 합성하였다. From the compound of Formula IX obtained in Scheme 1, a compound of Formula 2 was synthesized according to Scheme 2 below.
상기 방법으로 제조된 본 발명의 화합물을 하기 표 2에 나타내었다. The compounds of the present invention prepared by the above method are shown in Table 2 below.
[반응식 3]Scheme 3
상기 반응식 1에서 수득한 화학식 IX의 화합물로부터 하기 화학식 3의 화합물을 반응식 3에 따라 합성하였다. From the compound of Formula IX obtained in Scheme 1, a compound of Formula 3 was synthesized according to Scheme 3.
상기 방법으로 제조된 본 발명의 화합물을 하기 표 3에 나타내었다. The compounds of the present invention prepared by the above method are shown in Table 3 below.
[실시예 1]Example 1
4,5-비스(클로로메틸)-2-메틸피리딘-3-올 염산염 합성 (화학식 II)Synthesis of 4,5-bis (chloromethyl) -2-methylpyridin-3-ol hydrochloride (Formula II)
[4,5-bis(chloromethyl)-2-methylpyridin-3-ol hydrochloride] [4,5-bis (chloromethyl) -2-methylpyridin-3-ol hydrochloride]
피리독신염산염(화학식 I)(5 g)에 SOCl2(30 mL)와 DMF(0.2 mL)를 가한 후 80 ℃에서 3시간 환류교반하였다. 반응 혼합물을 실온으로 냉각한 후 Et2O를 가하고 빙냉 하에서 1시간 교반하였다. 석출된 고체를 감압여과하고 걸러진 고체를 Et2O로 세척한 후 건조시켜 목적 화합물 II (5.5 g)을 흰색의 고체로 얻었다.SOCl 2 (30 mL) and DMF (0.2 mL) were added to pyridoxine hydrochloride (Formula I) (5 g), followed by stirring under reflux at 80 ° C. for 3 hours. After cooling the reaction mixture to room temperature, Et 2 O was added and stirred under ice cooling for 1 hour. The precipitated solid was filtered under reduced pressure and the filtered solid was washed with Et 2 O and dried to obtain the target compound II (5.5 g) as a white solid.
1H-NMR ((CD3)2SO) δ 8.42 (s, 1H), 4.99 (s, 2H), 4.96 (s, 2H), 2.63 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 8.42 (s, 1H), 4.99 (s, 2H), 4.96 (s, 2H), 2.63 (s, 3H).
[실시예 2]Example 2
2,4,5-트라이메틸피리딘-3-올 합성 (화학식 III)Synthesis of 2,4,5-trimethylpyridin-3-ol (Formula III)
[2,4,5-trimethylpyridin-3-ol] [2,4,5-trimethylpyridin-3-ol]
화합물 II(10 g)의 아세트산(50 mL) 현탁액에 아연 분말 (8.08 g)을 소량씩 나누어 넣고 130 ℃에서 2시간 환류교반하였다. 반응혼합물을 실온으로 식힌 뒤 감압여과하고, 10 M NaOH 용액을 사용하여 여액의 pH를 6으로 조절하였다. 상기 여액을 소금으로 포화한 후 EtOAc로 추출하였다.A small amount of zinc powder (8.08 g) was added to an acetic acid (50 mL) suspension of compound II (10 g), and the mixture was stirred under reflux at 130 ° C. for 2 hours. The reaction mixture was cooled to room temperature and filtered under reduced pressure, and the pH of the filtrate was adjusted to 6 using 10 M NaOH solution. The filtrate was saturated with salt and extracted with EtOAc.
EtOAc 용액을 포화소금물로 세척하고 MgSO4로 건조, 여과하여 감압 농축하였다. 잔사를 관크로마토그래피(CHCl3:MeOH=20:1)로 정제하여 목적 화합물 III (5.2 g)를 흰색의 고체로 얻었다.The EtOAc solution was washed with saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (CHCl 3 : MeOH = 20: 1) to give the title compound III (5.2 g) as a white solid.
1H-NMR ((CD3)2SO) δ 8.49 (s, 1H), 7.72 (s, 1H), 2,31 (s, 3H), 2.12 (s, 3H), 2.08 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 8.49 (s, 1H), 7.72 (s, 1H), 2,31 (s, 3H), 2.12 (s, 3H), 2.08 (s, 3H).
[실시예 3]Example 3
6-브로모-2,4,5-트라이메틸피리딘-3-올 합성 (화학식 IV)Synthesis of 6-bromo-2,4,5-trimethylpyridin-3-ol (Formula IV)
[6-bromo-2,4,5-trimethylpyridin-3-ol] [6-bromo-2,4,5-trimethylpyridin-3-ol]
화합물 III(2.5 g)의 THF(30 mL) 현탁액에 1,3-디브로모-5,5-디메틸히단토인(DBDMH, 2.5 g)을 가한 뒤 실온에서 3시간 교반하였다. 반응혼합물을 농축한 후 잔사를 EtOAc과 물로 희석하고 수층을 EtOAc로 추출하였다. EtOAc 용액을 포화소금물로 세척하고 MgSO4로 건조, 여과하여 감압 농축하였다. 잔사를 관크로마토그래피(EtOAc:Hex=1:4)로 정제하여 목적 화합물 IV (3.22 g)을 연노란색의 고체로 얻었다.To a THF (30 mL) suspension of compound III (2.5 g) was added 1,3-dibromo-5,5-dimethylhydantoin (DBDMH, 2.5 g) and stirred at room temperature for 3 hours. After the reaction mixture was concentrated, the residue was diluted with EtOAc and water, and the aqueous layer was extracted with EtOAc. The EtOAc solution was washed with saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (EtOAc: Hex = 1: 4) to afford the desired compound IV (3.22 g) as a pale yellow solid.
1H-NMR (CDCl3) δ 5.56 (br s, 1H), 2.42 (s, 3H), 2.31 (s, 3H), 2.25 (s, 3H). 1 H-NMR (CDCl 3 ) δ 5.56 (br s, 1H), 2.42 (s, 3H), 2.31 (s, 3H), 2.25 (s, 3H).
[실시예 4]Example 4
2-브로모-5-((t-부틸다이페닐실릴)옥시)-3,4,6-트라이메틸피리딘 (화학식 V)2-bromo-5-((t-butyldiphenylsilyl) oxy) -3,4,6-trimethylpyridine (Formula V)
[2-bromo-5-((tert-butyldiphenylsilyl)oxy)-3,4,6-trimethylpyridine] [2-bromo-5-((tert-butyldiphenylsilyl) oxy) -3,4,6-trimethylpyridine]
화합물 IV(5.11 g)의 DMF(34 mL) 용액에 이미다졸(3.54 g)과 TBDPSCl(9.1 mL)를 가한 후 실온에서 16시간 교반하였다. 반응 혼합물에 물을 가한 후 CH2Cl2로 추출하였다. 추출액을 물, 포화소금물로 차례로 세척한 후, MgSO4로 건조, 여과한 후 여액을 감압 농축하였다. 잔사를 관크로마토그래피(hexanes 중 1% EtOAc)로 정제하여 목적 화합물 V (9.73 g)를 흰색의 고체로 얻었다.Imidazole (3.54 g) and TBDPSCl (9.1 mL) were added to a solution of Compound IV (5.11 g) in DMF (34 mL), followed by stirring at room temperature for 16 hours. Water was added to the reaction mixture, which was then extracted with CH 2 Cl 2 . The extract was washed successively with water and brine, dried over MgSO 4 , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by tube chromatography (1% EtOAc in hexanes) to afford the desired compound V (9.73 g) as a white solid.
1H-NMR (CDCl3) δ 7.67-7.62 (m, 4H), 7.44-7.31 (m, 6H), 2.21 (s, 3H), 2.16 (s, 3H), 1.98 (s, 3H), 1.10 (s, 9H). 1 H-NMR (CDCl 3 ) δ 7.67-7.62 (m, 4H), 7.44-7.31 (m, 6H), 2.21 (s, 3H), 2.16 (s, 3H), 1.98 (s, 3H), 1.10 ( s, 9H).
[실시예 5]Example 5
N-(5-((t-부틸다이페닐실릴)옥시)-3,4,6-트리메틸피리딘-2-일)-1.1-다이페닐메탄이민 (화학식 VI)N- (5-((t-butyldiphenylsilyl) oxy) -3,4,6-trimethylpyridin-2-yl) -1.1-diphenylmethaneimine (Formula VI)
[N-(5-((tert-butyldiphenylsilyl)oxy)-3,4,6-trimethylpyridin-2-yl)-1,1-diphenylmethanimine] [N- (5-((tert-butyldiphenylsilyl) oxy) -3,4,6-trimethylpyridin-2-yl) -1,1-diphenylmethanimine]
화합물 V(3.7 g)의 톨루엔(32 mL) 용액에 Pd2(dba)3(375 mg), BINAP(504 mg), NaOtBu(860 mg), 벤조페논이민(1.37 mL)을 차례대로 넣고 3시간 동안 아르곤 기류 하 환류교반하였다. 실온으로 식힌 후, 반응 혼합물을 EtOAc로 희석하고 물, 포화소금물로 세척하였다. EtOAc 용액을 MgSO4로 건조하고 여과한 후, 여액을 감압 농축하였다. 잔사를 관크로마토그래피(hexanes 중 3% 내지 5% EtOAc)로 정제하여 목적 화합물 VI(3.72 g)을 노란색 고체로 얻었다.To a solution of compound V (3.7 g) in toluene (32 mL) was added Pd 2 (dba) 3 (375 mg), BINAP (504 mg), NaO t Bu (860 mg), and benzophenoneimine (1.37 mL) in this order. It was stirred at reflux under argon for 3 hours. After cooling to rt, the reaction mixture was diluted with EtOAc and washed with water, saturated brine. The EtOAc solution was dried over MgSO 4 , filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (3% to 5% EtOAc in hexanes) to afford the desired compound VI (3.72 g) as a yellow solid.
1H-NMR (CDCl3) δ 7.85-7.75 (m, 2H), 7.66-7.61 (m, 4H), 7.48-7.28 (m, 11H), 7.25-7.10 (m, 3H), 2.09 (s, 3H), 1.84 (d, J = 9.1 Hz, 6H), 1.10 (s, 9H). 1 H-NMR (CDCl 3 ) δ 7.85-7.75 (m, 2H), 7.66-7.61 (m, 4H), 7.48-7.28 (m, 11H), 7.25-7.10 (m, 3H), 2.09 (s, 3H ), 1.84 (d, J = 9.1 Hz, 6H), 1.10 (s, 9H).
[실시예 6]Example 6
5-((t-부틸다이페닐실릴)옥시)-3,4,6-트리메틸피리딘-2-아민 (화학식 VII)5-((t-butyldiphenylsilyl) oxy) -3,4,6-trimethylpyridin-2-amine (Formula VII)
[5-((tert-butyldiphenylsilyl)oxy)-3,4,6-trimethylpyridin-2-amine] [5-((tert-butyldiphenylsilyl) oxy) -3,4,6-trimethylpyridin-2-amine]
화합물 VI(6.78 g)을 MeOH(60 mL)과 THF(7 mL)의 혼합 용매에 녹인 후 용액을 빙냉하였다. 여기에 AcCl(1.92 mL)를 천천히 적가한 후 실온으로 온도를 올려서 28시간 교반하였다. 반응 혼합물을 감압 농축한 후, 잔사를 EtOAc로 희석하고 2M NaOH를 가하여 용액의 pH를 7로 조절하였다. 이 용액을 포화소금물로 세척하고 MgSO4로 건조, 여과한 후 감압 농축하였다. 잔사를 관크로마토그래피(CH2Cl2 중 2.5% MeOH)로 정제하여 목적 화합물 VII(2.52 g)을 갈색 고체로 얻었다.Compound VI (6.78 g) was dissolved in a mixed solvent of MeOH (60 mL) and THF (7 mL) and the solution was ice cooled. AcCl (1.92 mL) was slowly added dropwise thereto, and the temperature was raised to room temperature and stirred for 28 hours. After the reaction mixture was concentrated under reduced pressure, the residue was diluted with EtOAc and 2M NaOH was added to adjust the pH of the solution to 7. The solution was washed with saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by tube chromatography (2.5% MeOH in CH 2 Cl 2 ) to afford the desired compound VII (2.52 g) as a brown solid.
1H-NMR ((CD3)2SO) δ 7.65 (d, J = 1.4 Hz, 4H), 7.48-7.36 (m, 6H), 5.02 (s, 2H), 1.92 (s, 3H), 1.86 (d, J = 2.4 Hz, 6H), 1.04 (s, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 7.65 (d, J = 1.4 Hz, 4H), 7.48-7.36 (m, 6H), 5.02 (s, 2H), 1.92 (s, 3H), 1.86 ( d, J = 2.4 Hz, 6H), 1.04 (s, 9H).
[실시예 7]Example 7
2,4,5-트라이메틸-6-((5-R-1,3,4-티아디아졸-2-일)아미노)피리딘-3-올 (화학식 2)의 일반적 합성법General synthesis of 2,4,5-trimethyl-6-((5-R-1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol (Formula 2)
[2,4,5-trimethyl-6-((5-R-1,3,4-thiadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5-R-1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol]
화합물 VII(1.0 mmol)과 다양한 치환기 R을 갖는 화합물 VIII(1.1 mmol)의 CH2Cl2(5 mL) 용액을 실온에서 교반하였다. 반응 혼합물을 감압 농축한 후, 잔사를 관크로마토그래피로 정제하여 화합물 IX를 얻었다.A solution of CH 2 Cl 2 (5 mL) of compound VII (1.0 mmol) and compound VIII (1.1 mmol) with various substituents R was stirred at room temperature. After the reaction mixture was concentrated under reduced pressure, the residue was purified by column chromatography to obtain Compound IX.
화합물 IX(1.0 mmol)를 N-메틸피롤리딘(10 mL)에 녹이고 Et3N(2.4 mmol), p-TsCl(1.2 mmol)을 차례대로 가한 후 실온에서 교반하였다. 반응 혼합물을 EtOAc로 희석하여 물, 포화소금물로 세척한 후, MgSO4로 건조, 여과하고 감압 농축하였다. 잔사를 관크로마토그래피로 정제하여 화합물 X를 얻었다.Compound IX (1.0 mmol) was dissolved in N-methylpyrrolidine (10 mL), and Et 3 N (2.4 mmol) and p-TsCl (1.2 mmol) were added sequentially, followed by stirring at room temperature. The reaction mixture was diluted with EtOAc, washed with water and saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give compound X.
화합물 X(1 mmol)를 THF(5 mL)에 녹인 후 1M n-Bu4NF THF 용액(1.2 mmol)을 가하고 실온에서 교반하였다. 반응 혼합물을 EtOAc로 희석하고 물, 포화소금물로 세척한 후, MgSO4로 건조, 여과하여 감압 농축하였다. 잔사를 관크로마토그래피로 정제하여 화합물 2를 얻었다. Compound X (1 mmol) was dissolved in THF (5 mL), 1M n-Bu 4 NF THF solution (1.2 mmol) was added thereto, and the mixture was stirred at room temperature. The reaction mixture was diluted with EtOAc, washed with water and saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give compound 2.
[실시예 8]Example 8
2,4,5-트라이메틸-6-((5-R-1,3,4-옥사다이아졸-2-일)아미노)피리딘-3-올 (화학식 3)의 일반적 합성법General synthesis of 2,4,5-trimethyl-6-((5-R-1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol (Formula 3)
[2,4,5-trimethyl-6-((5-R-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5-R-1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
화합물 VII(1.0 mmol)과 다양한 치환기 R을 갖는 화합물 VIII(1.1 mmol)의 CH2Cl2(5 mL) 용액을 실온에서 교반하였다. 반응 혼합물을 감압 농축한 후, 잔사를 관크로마토그래피로 정제하여 화합물 IX를 얻었다.A solution of CH 2 Cl 2 (5 mL) of compound VII (1.0 mmol) and compound VIII (1.1 mmol) with various substituents R was stirred at room temperature. After the reaction mixture was concentrated under reduced pressure, the residue was purified by column chromatography to obtain Compound IX.
화합물 IX(1.0 mmol)을 DMSO(5 mL)에 녹인 후, EDCI(1.2 mmol)를 가하고 50~60 ℃에서 교반하였다. 반응 혼합물을 실온으로 냉각한 후 EtOAc로 희석하였다. EtOAc 용액을 물, 포화소금물로 차례대로 세척한 후, MgSO4로 건조, 여과하고 감압 농축하였다. 잔사를 관크로마토그래피로 정제하여 화합물 XI를 얻었다.Compound IX (1.0 mmol) was dissolved in DMSO (5 mL), EDCI (1.2 mmol) was added thereto, and the obtained mixture was stirred at 50 to 60 ° C. The reaction mixture was cooled to rt and diluted with EtOAc. The EtOAc solution was washed sequentially with water and saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give compound XI.
화합물 XI(1 mmol)를 THF(5 mL)에 녹인 후 1M n-Bu4NF THF 용액(1.2 mmol)을 가하고 실온에서 교반하였다. 반응 혼합물을 EtOAc로 희석하고 물, 포화소금물로 세척한 후, MgSO4로 건조, 여과하여 감압 농축하였다. 잔사를 관크로마토그래피로 정제하여 화합물 3을 얻었다. Compound XI (1 mmol) was dissolved in THF (5 mL), then 1M n-Bu 4 NF THF solution (1.2 mmol) was added and stirred at room temperature. The reaction mixture was diluted with EtOAc, washed with water and saturated brine, dried over MgSO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give compound 3.
[실시예 9]Example 9
2,4,5-트라이메틸-6-((5-페닐-1,3,4-티아디아졸-2-일)아미노)피리딘-3-올 (화학식 2-036)2,4,5-trimethyl-6-((5-phenyl-1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol (Formula 2-036)
[2,4,5-trimethyl-6-((5-phenyl-1,3,4-thiadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5-phenyl-1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.55 (s, 1H), 8.29 (s, 1H), 7.96-7.87 (m, 2H), 7.56-7.43 (m, 3H), 2.43 (s, 3H), 2.23 (s, 3H), 2.17 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.55 (s, 1H), 8.29 (s, 1H), 7.96-7.87 (m, 2H), 7.56-7.43 (m, 3H), 2.43 (s, 3H ), 2.23 (s, 3H), 2.17 (s, 3H).
[실시예 10]Example 10
2,4,5-트라이메틸-6-((5-(p-톨일)-1,3,4-티아디아졸-2-일)아미노)피리딘-3-올 (화학식 2-037)2,4,5-trimethyl-6-((5- (p-tolyl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol (Formula 2-037)
[2,4,5-trimethyl-6-((5-(p-tolyl)-1,3,4-thiadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (p-tolyl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.50 (s, 1H), 8.28 (s, 1H), 7.79 (d, J = 8.1 Hz, 2H), 7.31 (d, J = 8.0 Hz, 2H), 2.42 (s, 3H), 2.36 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.50 (s, 1H), 8.28 (s, 1H), 7.79 (d, J = 8.1 Hz, 2H), 7.31 (d, J = 8.0 Hz, 2H) , 2.42 (s, 3H), 2.36 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H).
[실시예 11]Example 11
6-((5-(4-브로모페닐)-1,3,4-티아디아졸-2-일)아미노)-2,4,5-트라이메틸피리딘-3-올 (화학식 2-039)6-((5- (4-bromophenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol (Formula 2-039)
[6-((5-(4-bromophenyl)-1,3,4-thiadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (4-bromophenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.61 (s, 1H), 8.31 (s, 1H), 7.86 (d, J = 8.5 Hz, 2H), 7.73-7.68 (m, 2H), 2.43 (s, 3H), 2.23 (s, 3H), 2.17 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.61 (s, 1H), 8.31 (s, 1H), 7.86 (d, J = 8.5 Hz, 2H), 7.73-7.68 (m, 2H), 2.43 ( s, 3H), 2.23 (s, 3H), 2.17 (s, 3H).
[실시예 12]Example 12
6-((5-(4-(디메틸아미노)페닐)-1,3,4-티아디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 2-041)6-((5- (4- (dimethylamino) phenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol (Formula 2-041 )
[6-((5-(4-(dimethylamino)phenyl)-1,3,4-thiadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (4- (dimethylamino) phenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.29 (s, 1H), 8.23 (s, 1H), 7.70 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 9.0 Hz, 2H), 2.98 (s, 6H), 2.41 (s, 3H), 2.21 (s, 3H), 2.16 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.29 (s, 1H), 8.23 (s, 1H), 7.70 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 9.0 Hz, 2H) , 2.98 (s, 6H), 2.41 (s, 3H), 2.21 (s, 3H), 2.16 (s, 3H).
[실시예 13]Example 13
6-((5-(4-메톡시페닐)-1,3,4-티아디아졸-2-일)아미노)-2,4,5-트라이메틸피리딘-3-올 (화학식 2-045)6-((5- (4-methoxyphenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol (Formula 2-045)
[6-((5-(4-methoxyphenyl)-1,3,4-thiadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (4-methoxyphenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.44 (s, 1H), 8.26 (s, 1H), 7.83 (d, J = 8.8 Hz, 2H), 7.09-7.03 (m, 2H), 3.82 (s, 3H), 2.42 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.44 (s, 1H), 8.26 (s, 1H), 7.83 (d, J = 8.8 Hz, 2H), 7.09-7.03 (m, 2H), 3.82 ( s, 3H), 2.42 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H).
[실시예 14]Example 14
2,4,5-트라이메틸-6-((5-(4-(트라이플루오로메틸)페닐)-1,3,4-티아디아졸-2-일)아미노)피리딘-3-올 (화학식 2-046)2,4,5-trimethyl-6-((5- (4- (trifluoromethyl) phenyl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol 2-046)
[2,4,5-trimethyl-6-((5-(4-(trifluoromethyl)phenyl)-1,3,4-thiadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (4- (trifluoromethyl) phenyl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.73 (s, 1H), 8.33 (s, 1H), 8.12 (d, J = 8.1 Hz, 2H), 7.85 (d, J = 8.2 Hz, 2H), 2.44 (s, 3H), 2.23 (s, 3H), 2.19-2.14 (m, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.73 (s, 1H), 8.33 (s, 1H), 8.12 (d, J = 8.1 Hz, 2H), 7.85 (d, J = 8.2 Hz, 2H) , 2.44 (s, 3H), 2.23 (s, 3H), 2.19-2.14 (m, 3H).
[실시예 15]Example 15
2,4,5-트리메틸-6-((5-(4-(트리플루오로메톡시)페닐)-1,3,4-티아디아졸-2-일)아미노)피리딘-3-올 (화학식 2-047)2,4,5-trimethyl-6-((5- (4- (trifluoromethoxy) phenyl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol -047)
[2,4,5-trimethyl-6-((5-(4-(trifluoromethoxy)phenyl)-1,3,4-thiadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (4- (trifluoromethoxy) phenyl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.63 (s, 1H), 8.31 (s, 1H), 8.04 (d, J = 8.8 Hz, 2H), 7.53-7.48 (m, 2H), 2.43 (s, 3H), 2.23 (s, 3H), 2.17 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.63 (s, 1H), 8.31 (s, 1H), 8.04 (d, J = 8.8 Hz, 2H), 7.53-7.48 (m, 2H), 2.43 ( s, 3H), 2.23 (s, 3H), 2.17 (s, 3H).
[실시예 16]Example 16
6-((5-(4-(벤질옥시)페닐)-1,3,4-티아디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 2-048)6-((5- (4- (benzyloxy) phenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol (Formula 2-048 )
[6-((5-(4-(benzyloxy)phenyl)-1,3,4-thiadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (4- (benzyloxy) phenyl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.43 (s, 1H), 8.26 (s, 1H), 7.83 (d, J = 8.7 Hz, 2H), 7.51-7.46 (m, 2H), 7.44-7.38 (m, 2H), 7.35 (dt, J = 9.6, 4.3 Hz, 1H), 7.17-7.12 (m, 2H), 5.18 (s, 2H), 2.42 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.43 (s, 1H), 8.26 (s, 1H), 7.83 (d, J = 8.7 Hz, 2H), 7.51-7.46 (m, 2H), 7.44- 7.38 (m, 2H), 7.35 (dt, J = 9.6, 4.3 Hz, 1H), 7.17-7.12 (m, 2H), 5.18 (s, 2H), 2.42 (s, 3H), 2.22 (s, 3H) , 2.16 (s, 3 H).
[실시예 17]Example 17
6-((5-(벤조[d][1,3]디옥솔-5-일)-1,3,4-티아디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 2-049)6-((5- (benzo [d] [1,3] dioxol-5-yl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridine- 3-ol (Formula 2-049)
[6-((5-(benzo[d][1,3]dioxo-5-yl)-1,3,4-thiadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (benzo [d] [1,3] dioxo-5-yl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol ]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.47 (s, 1H), 8.27 (s, 1H), 7.46 (d, J = 1.6 Hz, 1H), 7.36 (dd, J = 8.1, 1.7 Hz, 1H), 7.03 (d, J = 8.1 Hz, 1H), 6.11 (s, 2H), 2.42 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.47 (s, 1H), 8.27 (s, 1H), 7.46 (d, J = 1.6 Hz, 1H), 7.36 (dd, J = 8.1, 1.7 Hz, 1H), 7.03 (d, J = 8.1 Hz, 1H), 6.11 (s, 2H), 2.42 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H).
[실시예 18]Example 18
6-((5-([1,1'-비페닐]-4-일)-1,3,4-티아디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 2-053)6-((5-([1,1'-biphenyl] -4-yl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridine-3- All (Formula 2-053)
[6-((5-([1,1’-biphenyl]4-yl)-1,3,4-thiadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5-([1,1'-biphenyl] 4-yl) -1,3,4-thiadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.58 (s, 1H), 8.30 (s, 1H), 7.99 (d, J = 8.3 Hz, 2H), 7.84-7.79 (m, 2H), 7.77-7.72 (m, 2H), 7.51 (dd, J = 10.4, 4.8 Hz, 2H), 7.41 (t, J = 7.3 Hz, 1H), 2.45 (s, 3H), 2.24 (s, 3H), 2.17 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.58 (s, 1H), 8.30 (s, 1H), 7.99 (d, J = 8.3 Hz, 2H), 7.84-7.79 (m, 2H), 7.77- 7.72 (m, 2H), 7.51 (dd, J = 10.4, 4.8 Hz, 2H), 7.41 (t, J = 7.3 Hz, 1H), 2.45 (s, 3H), 2.24 (s, 3H), 2.17 (s , 3H).
[실시예 19]Example 19
2,4,5-트리메틸-6-((5-(나프탈렌-1-일)-1,3,4-티아디아졸-2-일)아미노)피리딘-3-올 (화학식 2-056)2,4,5-trimethyl-6-((5- (naphthalen-1-yl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol (Formula 2-056)
[2,4,5-trimethyl-6-((5-(naphthalene-1-yl)-1,3,4-thiadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (naphthalene-1-yl) -1,3,4-thiadiazol-2-yl) amino) pyridin-3-ol]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.62 (s, 1H), 8.76-8.69 (m, 1H), 8.30 (s, 1H), 8.11-8.03 (m, 2H), 7.85 (dd, J = 7.2, 1.1 Hz, 1H), 7.70-7.58 (m, 3H), 2.38 (s, 3H), 2.26 (s, 3H), 2.18 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.62 (s, 1H), 8.76-8.69 (m, 1H), 8.30 (s, 1H), 8.11-8.03 (m, 2H), 7.85 (dd, J = 7.2, 1.1 Hz, 1H), 7.70-7.58 (m, 3H), 2.38 (s, 3H), 2.26 (s, 3H), 2.18 (s, 3H).
[실시예 20]Example 20
4-(5-((5-히드록시-3,4,6-트리메틸피리딘-2-일)아미노)-1,3,4-티아디아졸-2-일)페닐 4-메틸벤젠설포네이트 (화학식 2-061)4- (5-((5-hydroxy-3,4,6-trimethylpyridin-2-yl) amino) -1,3,4-thiadiazol-2-yl) phenyl 4-methylbenzenesulfonate ( Formula 2-061)
[4-(5-((5-hydroxy-3,4,6-trimethylpyridin-2-yl)amino)-1,3,4-thiadiazol-2-yl)phenyl 4-methylbenzenesulfonate] [4- (5-((5-hydroxy-3,4,6-trimethylpyridin-2-yl) amino) -1,3,4-thiadiazol-2-yl) phenyl 4-methylbenzenesulfonate]
실시예 7의 방법에 따라 제조하였다.Prepared according to the method of Example 7.
1H-NMR ((CD3)2SO) δ 10.60 (s, 1H), 8.30 (s, 1H), 7.91 (d, J = 8.7 Hz, 2H), 7.81-7.75 (m, 2H), 7.53-7.46 (m, 2H), 7.18-7.12 (m, 2H), 2.43 (s, 3H), 2.41 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.60 (s, 1H), 8.30 (s, 1H), 7.91 (d, J = 8.7 Hz, 2H), 7.81-7.75 (m, 2H), 7.53- 7.46 (m, 2H), 7.18-7.12 (m, 2H), 2.43 (s, 3H), 2.41 (s, 3H), 2.22 (s, 3H), 2.16 (s, 3H).
[실시예 21]Example 21
2,4,5-트라이메틸-6-((5-메틸-1,3,4-옥사다이아졸-2-일)아미노)피리딘-3-올 (화학식 3-002)2,4,5-trimethyl-6-((5-methyl-1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol (Formula 3-002)
[2,4,5-trimethyl-6-((5-methyl-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5-methyl-1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.38 (s, 1H), 8.44 (s, 1H), 2.33 (s, 3H), 2.28 (s, 3H), 2.10 (d, J = 34.2 Hz, 6H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.38 (s, 1H), 8.44 (s, 1H), 2.33 (s, 3H), 2.28 (s, 3H), 2.10 (d, J = 34.2 Hz, 6H).
[실시예 22]Example 22
2,4,5-트라이메틸-6-((5-(트라이플루오로메틸)-1,3,4-옥사다이아졸-2-일)아미노)피리딘-3-올 (화학식 3-010)2,4,5-trimethyl-6-((5- (trifluoromethyl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol (Formula 3-010)
[2,4,5-trimethyl-6-((5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (trifluoromethyl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 10.44 (s, 1H), 8.67 (s, 1H), 2.33 (s, 3H), 2.17 (d, J = 21.1 Hz, 6H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.44 (s, 1H), 8.67 (s, 1H), 2.33 (s, 3H), 2.17 (d, J = 21.1 Hz, 6H).
[실시예 23]Example 23
2,4,5-트라이메틸-6-((5-페닐-1,3,4-옥사다이아졸-2-일)아미노)피리딘-3-올 (화학식 3-036)2,4,5-trimethyl-6-((5-phenyl-1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol (Formula 3-036)
[2,4,5-trimethyl-6-((5-phenyl-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5-phenyl-1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.79 (s, 1H), 8.53 (s, 1H), 7.91 (d, J = 57.9 Hz, 2H), 7.58-7.51 (m, 3H), 2.45-2.06 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.79 (s, 1H), 8.53 (s, 1H), 7.91 (d, J = 57.9 Hz, 2H), 7.58-7.51 (m, 3H), 2.45- 2.06 (m, 9 H).
[실시예 24]Example 24
6-((5-(4-(디메틸아미노)페닐)-1,3,4-옥사디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 3-041)6-((5- (4- (dimethylamino) phenyl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol (Formula 3-041 )
[6-((5-(4-(dimethylamino)phenyl)-1,3,4-oxadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (4- (dimethylamino) phenyl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.53 (s, 1H), 8.43 (s, 1H), 7.61 (d, J = 8.2 Hz, 2H), 6.80 (d, J = 8.8 Hz, 2H), 2.98 (s, 6H), 2.18 (dd, J = 47.2, 24.9 Hz, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.53 (s, 1H), 8.43 (s, 1H), 7.61 (d, J = 8.2 Hz, 2H), 6.80 (d, J = 8.8 Hz, 2H) , 2.98 (s, 6H), 2.18 (dd, J = 47.2, 24.9 Hz, 9H).
[실시예 25]Example 25
2,4,5-트라이메틸-6-((5-(4-트라이플루오로메틸)페닐)-1,3,4-옥사다이아졸-2-일)아미노)피리딘-3-올 (화학식 3-046)2,4,5-trimethyl-6-((5- (4-trifluoromethyl) phenyl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol -046)
[2,4,5-trimethyl-6-((5-(4-(trifluoromethyl)phenyl)-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (4- (trifluoromethyl) phenyl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.99 (s, 1H), 8.60 (s, 1H), 8.05 (s, 2H), 7.91 (d, J = 8.3 Hz, 2H), 2.34 (s, 3H), 2.18 (d, J = 16.9 Hz, 6H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.99 (s, 1H), 8.60 (s, 1H), 8.05 (s, 2H), 7.91 (d, J = 8.3 Hz, 2H), 2.34 (s, 3H), 2.18 (d, J = 16.9 Hz, 6H).
[실시예 26]Example 26
2,4,5-트리메틸-6-((5-(4-(트리플루오로메톡시)페닐)-1,3,4-옥사디아졸-2-일)아미노)피리딘-3-올 (화학식 3-047)2,4,5-trimethyl-6-((5- (4- (trifluoromethoxy) phenyl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol -047)
[2,4,5-trimethyl-6-((5-(4-(trifluoromethoxy)phenyl)-1,3,4-oxadazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (4- (trifluoromethoxy) phenyl) -1,3,4-oxadazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.88 (s, 1H), 8.54 (s, 1H), 7.96 (s, 2H), 7.55 (dd, J = 8.9, 0.9 Hz, 2H), 2.42-2.06 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.88 (s, 1H), 8.54 (s, 1H), 7.96 (s, 2H), 7.55 (dd, J = 8.9, 0.9 Hz, 2H), 2.42- 2.06 (m, 9 H).
[실시예 27]Example 27
6-((5-(4-(벤질옥시)페닐)-1,3,4-옥사디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 3-048)6-((5- (4- (benzyloxy) phenyl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol (Formula 3-048) )
[6-((5-(4-(benzyloxy)phenyl)-1,3,4-oxadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (4- (benzyloxy) phenyl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.68 (s, 1H), 8.49 (s, 1H), 7.78 (s, 2H), 7.51-7.32 (m, 5H), 7.20-7.15 (m, 2H), 5.19 (s, 2H), 2.40-2.06 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.68 (s, 1H), 8.49 (s, 1H), 7.78 (s, 2H), 7.51-7.32 (m, 5H), 7.20-7.15 (m, 2H ), 5.19 (s, 2 H), 2.40-2.06 (m, 9 H).
[실시예 28]Example 28
6-((5-(벤조[d][1,3]디옥솔-5-일)-1,3,4-옥사디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 3-049)6-((5- (benzo [d] [1,3] dioxol-5-yl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridine- 3-ol (Formula 3-049)
[6-((5-(benzo[d][1,3]dioxo-5-yl)-1,3,4-oxadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (benzo [d] [1,3] dioxo-5-yl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol ]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.71 (s, 1H), 8.47 (s, 1H), 7.46-7.26 (m, 2H), 7.07 (d, J = 8.1 Hz, 1H), 6.13 (s, 2H), 2.35-2.03 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.71 (s, 1H), 8.47 (s, 1H), 7.46-7.26 (m, 2H), 7.07 (d, J = 8.1 Hz, 1H), 6.13 ( s, 2H), 2.35-2.03 (m, 9H).
[실시예 29]Example 29
6-((5-([1,1'-비페닐]-4-일)-1,3,4-옥사디아졸-2-일)아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 3-053)6-((5-([1,1'-biphenyl] -4-yl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridine-3- All (Formula 3-053)
[6-((5-([1,1’-biphenyl]4-yl)-1,3,4-oxadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5-([1,1'-biphenyl] 4-yl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.83 (s, 1H), 8.54 (s, 1H), 8.04-7.81 (m, 4H), 7.78-7.70 (m, 2H), 7.55-7.46 (m, 2H), 7.44-7.38 (m, 1H), 2.31 (s, 3H), 2.26-2.01 (m, 6H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.83 (s, 1H), 8.54 (s, 1H), 8.04-7.81 (m, 4H), 7.78-7.70 (m, 2H), 7.55-7.46 (m , 2H), 7.44-7.38 (m, 1 H), 2.31 (s, 3 H), 2.26-2.01 (m, 6H).
[실시예 30]Example 30
2,4,5-트리메틸-6-((5-(나프탈렌-1-일)-1,3,4-옥사디아졸-2-일)아미노)피리딘-3-올 (화학식 3-056)2,4,5-trimethyl-6-((5- (naphthalen-1-yl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol (Formula 3-056)
[2,4,5-trimethyl-6-((5-(naphthalene-1-yl)-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (naphthalene-1-yl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.92 (s, 1H), 9.05 (d, J = 8.1 Hz, 1H), 8.50 (s, 1H), 8.16-7.94 (m, 4H), 7.75-7.61 (m, 4H), 2.34 (s, 3H), 2.16 (d, J = 9.3 Hz, 6H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.92 (s, 1H), 9.05 (d, J = 8.1 Hz, 1H), 8.50 (s, 1H), 8.16-7.94 (m, 4H), 7.75- 7.61 (m, 4 H), 2.34 (s, 3 H), 2.16 (d, J = 9.3 Hz, 6 H).
[실시예 31]Example 31
4-(5-((5-히드록시-3,4,6-트리메틸피리딘-2-일)아미노)-1,3,4-옥사디아졸-2-일)페닐 4-메틸벤젠설포네이트 (화학식 3-061)4- (5-((5-hydroxy-3,4,6-trimethylpyridin-2-yl) amino) -1,3,4-oxadiazol-2-yl) phenyl 4-methylbenzenesulfonate ( Formula 3-061)
[4-(5-((5-hydroxy-3,4,6-trimethylpyridin-2-yl)amino)-1,3,4-oxadiazol-2-yl)phenyl 4-methylbenzenesulfonate] [4- (5-((5-hydroxy-3,4,6-trimethylpyridin-2-yl) amino) -1,3,4-oxadiazol-2-yl) phenyl 4-methylbenzenesulfonate]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.84 (s, 1H), 8.50 (s, 1H), 7.92-7.72 (m, 4H), 7.49 (d, J = 8.1 Hz, 2H), 7.24-7.18 (m, 2H), 2.43 (s, 3H), 2.34-2.02 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.84 (s, 1H), 8.50 (s, 1H), 7.92-7.72 (m, 4H), 7.49 (d, J = 8.1 Hz, 2H), 7.24- 7.18 (m, 2 H), 2.43 (s, 3 H), 2.34-2.02 (m, 9 H).
[실시예 32]Example 32
2,4,5-트리메틸-6-(5-(피리딘-4-일)-1,3,4-옥사디아졸-2-일아미노)피리딘-3-올 (화학식 3-065)2,4,5-trimethyl-6- (5- (pyridin-4-yl) -1,3,4-oxadiazol-2-ylamino) pyridin-3-ol (Formula 3-065)
[2,4,5-trimethyl-6-((5-(pyridin-4-yl)-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (pyridin-4-yl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 10.16-9.95 (m, 1H), 8.77-8.73 (m, 2H), 8.62-8.48 (m, 1H), 7.87-7.70 (m, 2H), 2.32-1.96 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.16-9.95 (m, 1H), 8.77-8.73 (m, 2H), 8.62-8.48 (m, 1H), 7.87-7.70 (m, 2H), 2.32 -1.96 (m, 9H).
[실시예 33]Example 33
2,4,5-트리메틸-6-((5-(피리미딘-4-일)-1,3,4-옥사디아졸-2-일)아미노)피리딘-3-올 (화학식 3-073)2,4,5-trimethyl-6-((5- (pyrimidin-4-yl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol (Formula 3-073)
[2,4,5-trimethyl-6-((5-(pyrimidin-4-yl)-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (pyrimidin-4-yl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 10.21 (s, 1H), 9.30 (s, 1H), 8.98 (d, J = 5.3 Hz, 1H), 8.61 (s, 1H), 8.03 (s, 1H), 2.37-2.02 (m, 9H). 1 H-NMR ((CD 3 ) 2 SO) δ 10.21 (s, 1H), 9.30 (s, 1H), 8.98 (d, J = 5.3 Hz, 1H), 8.61 (s, 1H), 8.03 (s, 1H), 2.37-2.02 (m, 9H).
[실시예 34]Example 34
2,4,5-트리메틸-6-(5-(5-메틸-1H-피라졸-3-일)-1,3,4-옥사디아졸-2-일아미노)피리딘-3-올 (화학식 3-086)2,4,5-trimethyl-6- (5- (5-methyl-1H-pyrazol-3-yl) -1,3,4-oxadiazol-2-ylamino) pyridin-3-ol 3-086)
[2,4,5-trimethyl-6-((5-(5-methyl-1H-pyrazol-3-yl)-1,3,4-oxadiazol-2-yl)amino)pyridin-3-ol] [2,4,5-trimethyl-6-((5- (5-methyl-1H-pyrazol-3-yl) -1,3,4-oxadiazol-2-yl) amino) pyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 13.07 (s, 1H), 9.65 (s, 1H), 8.48 (s, 1H), 6.46 (s, 1H), 2.28 (s, 3H), 2.27 (s, 3H), 2.15 (s, 3H), 2.08 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 13.07 (s, 1H), 9.65 (s, 1H), 8.48 (s, 1H), 6.46 (s, 1H), 2.28 (s, 3H), 2.27 ( s, 3H), 2.15 (s, 3H), 2.08 (s, 3H).
[실시예 35]Example 35
6-(5-(1H-인돌-3-일)-1,3,4-옥사디아졸-2-일아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 3-100)6- (5- (1H-indol-3-yl) -1,3,4-oxadiazol-2-ylamino) -2,4,5-trimethylpyridin-3-ol (Formula 3-100)
[6-((5-(1H-indol-3-yl)-1,3,4-oxadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (1H-indol-3-yl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin-3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 11.79 (s, 1H), 9.53 (s, 1H), 8.41 (s, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.90 (d, J = 2.7 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.20 (dd, J = 15.9, 7.5 Hz, 2H), 2.29 (s, 3H), 2.17 (s, 3H), 2.12 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 11.79 (s, 1H), 9.53 (s, 1H), 8.41 (s, 1H), 8.02 (d, J = 7.6 Hz, 1H), 7.90 (d, J = 2.7 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 7.20 (dd, J = 15.9, 7.5 Hz, 2H), 2.29 (s, 3H), 2.17 (s, 3H), 2.12 ( s, 3H).
[실시예 36]Example 36
6-(5-(2,3-디히드로벤조[b][1,4]다이옥신-2-일)-1,3,4-옥사디아졸-2-일아미노)-2,4,5-트리메틸피리딘-3-올 (화학식 3-123)6- (5- (2,3-dihydrobenzo [b] [1,4] dioxin-2-yl) -1,3,4-oxadiazol-2-ylamino) -2,4,5- Trimethylpyridin-3-ol (Formula 3-123)
[6-((5-(2,3-dihydrobenzo[b][1,4]dioxin-2-yl)-1,3,4-oxadiazol-2-yl)amino)-2,4,5-trimethylpyridin-3-ol] [6-((5- (2,3-dihydrobenzo [b] [1,4] dioxin-2-yl) -1,3,4-oxadiazol-2-yl) amino) -2,4,5-trimethylpyridin -3-ol]
실시예 8의 방법에 따라 제조하였다.Prepared according to the method of Example 8.
1H-NMR ((CD3)2SO) δ 9.78 (s, 1H), 8.49 (s, 1H), 6.92-6.86 (m, 4H), 5.62-5.60 (m, 1H), 4.56 (dd, J = 11.8, 2.3 Hz, 1H), 4.42 (dd, J = 11.8, 5.8 Hz, 1H), 2.25 (s, 3H), 2.13 (s, 3H), 2.03 (s, 3H). 1 H-NMR ((CD 3 ) 2 SO) δ 9.78 (s, 1H), 8.49 (s, 1H), 6.92-6.86 (m, 4H), 5.62-5.60 (m, 1H), 4.56 (dd, J = 11.8, 2.3 Hz, 1H), 4.42 (dd, J = 11.8, 5.8 Hz, 1H), 2.25 (s, 3H), 2.13 (s, 3H), 2.03 (s, 3H).
[시험예 1][Test Example 1]
TNF-α 또는 IL-6로 유도한 단핵구의 장상피세포 부착 억제 활성 시험Inhibitory activity test of TNF-α or IL-6 induced monocyte epithelial cell adhesion
<시험 방법><Test method>
HT-29 사람 대장암 유래 상피세포와 U937 사람 유래 단핵구성 세포는 10% FBS, 1% penicillin/streptomycin(PS)가 함유된 RPMI 1640 배지로 37℃, 5% CO2 조건 하에서 배양하여, 세포가 배양 플라스크에 80% 이상 성장하면 1:3의 비율로 계대하여 본 실험에 사용하였다. HT-29 세포를 24 well plate에 2×105 cells/cm2의 농도로 배양하여 1% FBS와 1% PS만 함유된 배지에 시험 약물을 1시간 전처리 하였다. 그 후 10 μg/mL BCECF-AM를 처리하여 37℃에서 30분 반응시켜 BCECF가 세포 내에 탑재된 U937 세포와 TNF-α(10 ng/mL) 또는 IL-6(10 ng/mL)를 앞서 시험 약물이 처리된 HT-29 세포와 37℃에서 3시간 반응시켰다. 반응이 끝나면 배지를 제거하고 부착되지 않은 U937 세포를 제거하기 위하여 PBS로 2회 세척하였다. 다음 단계로 세포 용해를 위하여 0.1% Triton X-100 (0.1 M Tris)를 처리하여 실온에서 30분 반응시킨 후, Fluostar optima microplate reader (BMG Labtechnologies, Germany)을 사용하여 형광을 측정하여 정량하였다 (Carvalho et al., 1996; Thapa et al., 2008). HT-29 human colon cancer-derived epithelial cells and U937 human-derived mononuclear cells were cultured under RPMC 1640 medium containing 10% FBS and 1% penicillin / streptomycin (PS) under 37 ° C. and 5% CO 2 . When more than 80% growth in the culture flask was passaged at a ratio of 1: 3 was used in this experiment. HT-29 cells were incubated in a 24 well plate at a concentration of 2 × 10 5 cells / cm 2 , and the test drug was pretreated for 1 hour in a medium containing only 1% FBS and 1% PS. Thereafter, 10 μg / mL BCECF-AM was treated and reacted at 37 ° C. for 30 minutes to previously test U937 cells loaded with BCECF and TNF-α (10 ng / mL) or IL-6 (10 ng / mL). The drug-treated HT-29 cells were reacted for 3 hours at 37 ° C. At the end of the reaction, the medium was removed and washed twice with PBS to remove unattached U937 cells. In the next step, the cells were treated with 0.1% Triton X-100 (0.1 M Tris) for 30 minutes at room temperature, followed by fluorescence measurement using a Fluostar optima microplate reader (BMG Labtechnologies, Germany) (Carvalho et al., 1996; Thapa et al., 2008).
<TNF-α로 유도한 단핵구의 장상피세포 부착 억제 활성>Inhibitory Activity of TNF-α Induced Monocyte Epithelial Adhesion
TNF-α에 의해 유도된 장 상피 세포(HT-29)와 단핵구성 세포(U937)의 부착에 대한 시험 약물(1μM)의 억제 활성을 조사한 결과를 하기 표 4에 나타내었다. The inhibitory activity of the test drug (1 μM) on the adhesion of TNF-α induced intestinal epithelial cells (HT-29) and mononuclear cells (U937) is shown in Table 4 below.
| 화합물compound | 화합물 농도 (μM)Compound concentration (μM) | 억제 활성(%)Inhibitory Activity (%) |
| 5-ASA5-ASA | 10001000 | 9.9±2.1* 9.9 ± 2.1 * |
| 5-ASA5-ASA | 1One | 3.8±1.03.8 ± 1.0 |
| 2-0362-036 | 1One | 78.7±2.5* 78.7 ± 2.5 * |
| 2-0372-037 | 1One | 16.6±3.1* 16.6 ± 3.1 * |
| 2-0392-039 | 1One | 13.7±2.3* 13.7 ± 2.3 * |
| 2-0412-041 | 1One | 62.5±6.5* 62.5 ± 6.5 * |
| 2-0452-045 | 1One | 13.2±1.9* 13.2 ± 1.9 * |
| 2-0462-046 | 1One | 52.3±1.1* 52.3 ± 1.1 * |
| 2-0472-047 | 1One | 30.9±6.9* 30.9 ± 6.9 * |
| 2-0482-048 | 1One | 19.5±3.319.5 ± 3.3 |
| 2-0492-049 | 1One | 19.4±6.619.4 ± 6.6 |
| 2-0532-053 | 1One | 38.4±2.6* 38.4 ± 2.6 * |
| 2-0562-056 | 1One | 86.8±3.5* 86.8 ± 3.5 * |
| 2-0612-061 | 1One | 54.6±3.9* 54.6 ± 3.9 * |
| 3-0023-002 | 1One | 26.2±4.6* 26.2 ± 4.6 * |
| 3-0103-010 | 1One | 35.8±5.0* 35.8 ± 5.0 * |
| 3-0363-036 | 1One | 1.4±3.01.4 ± 3.0 |
| 3-0413-041 | 1One | 15.3±4.215.3 ± 4.2 |
| 3-0463-046 | 1One | 55.7±4.9* 55.7 ± 4.9 * |
| 3-0473-047 | 1One | 60.3±6.5* 60.3 ± 6.5 * |
| 3-0483-048 | 1One | 13.6±2.513.6 ± 2.5 |
| 3-0493-049 | 1One | 45.0±5.1* 45.0 ± 5.1 * |
| 3-0533-053 | 1One | 22.5±4.822.5 ± 4.8 |
| 3-0563-056 | 1One | 81.4±3.0* 81.4 ± 3.0 * |
| 3-0613-061 | 1One | 32.3±3.9* 32.3 ± 3.9 * |
| 3-0653-065 | 1One | 61.9±3.9* 61.9 ± 3.9 * |
| 3-0733-073 | 1One | 87.2±4.1* 87.2 ± 4.1 * |
| 3-0863-086 | 1One | 24.2±2.824.2 ± 2.8 |
| 3-1003-100 | 1One | 28.3±4.228.3 ± 4.2 |
| 3-1233-123 | 1One | 27.4±6.1* 27.4 ± 6.1 * |
* P <0.05 compared to vehicle-treated control group.* P <0.05 compared to vehicle-treated control group.
상기 표 4에 나타낸 바와 같이, 현재 임상에서 염증성 장질환 치료에 쓰이고 있는 약물인 설파살라진의 활성 대사체인 5-ASA(양성대조군)의 경우는 1000μM 농도에서 9.9%의 억제율을 나타내어 화합물들의 시험 농도인 1μM의 1000배 높은 농도에서도 그 효과가 매우 미미한 반면, 3-073 화합물과 2-056 화합물은 약 87%의 억제율을 나타내었고, 3-056 화합물은 약 81%의 억제율을 나타내었으며, 2-036 화합물은 약 79%의 억제율을 나타내어 매우 우수한 활성을 보였다. 화합물 2-041, 2-046, 2-061, 3-046을 비롯한 대부분의 화합물이 대조 물질인 5-ASA에 비해서 우수한 억제 활성을 갖는 것으로 확인되었다. As shown in Table 4, 5-ASA (positive control), an active metabolite of sulfasalazine, a drug currently used in the treatment of inflammatory bowel disease, exhibits an inhibition rate of 9.9% at a concentration of 1000 μM. At 1000-fold higher concentration of, the effect was very small, while 3-073 compound and 2-056 compound showed about 87% inhibition rate, 3-056 compound showed about 81% inhibition rate, and 2-036 compound Showed an inhibition rate of about 79%, showing very good activity. Most compounds including compounds 2-041, 2-046, 2-061, 3-046 were found to have superior inhibitory activity compared to the 5-ASA control.
<IL-6로 유도한 단핵구의 장상피세포 부착 억제 활성>IL-6-induced monocyte adhesion inhibitory activity of monocytes
상기와 같은 방법으로, IL-6에 의해 유도된 장 상피 세포(HT-29)와 단핵구성 세포(U937)의 부착에 대한 시험 약물(1μM)의 억제 활성을 조사한 결과를 하기 표 5에 나타내었다. In the same manner as described above, the results of examining the inhibitory activity of the test drug (1 μM) on the adhesion of IL-6-induced intestinal epithelial cells (HT-29) and mononuclear cells (U937) are shown in Table 5 below. .
| 화합물compound | 화합물 농도 (μM)Compound concentration (μM) | 억제 활성(%)Inhibitory Activity (%) |
| 5-ASA5-ASA | 10001000 | 1.7±7.81.7 ± 7.8 |
| 2-0362-036 | 1One | 68.6±5.4* 68.6 ± 5.4 * |
| 2-0372-037 | 1One | 13.1±7.1* 13.1 ± 7.1 * |
| 2-0392-039 | 1One | 9.3±12.3* 9.3 ± 12.3 * |
| 2-0412-041 | 1One | 51.7±4.8* 51.7 ± 4.8 * |
| 2-0452-045 | 1One | 8.7±6.3* 8.7 ± 6.3 * |
| 2-0462-046 | 1One | 50.4±5.4* 50.4 ± 5.4 * |
| 2-0472-047 | 1One | 21.4±5.821.4 ± 5.8 |
| 2-0482-048 | 1One | 9.6±5.59.6 ± 5.5 |
| 2-0492-049 | 1One | 11.4±4.411.4 ± 4.4 |
| 2-0532-053 | 1One | 30.5±4.9* 30.5 ± 4.9 * |
| 2-0562-056 | 1One | 77.7±3.2* 77.7 ± 3.2 * |
| 2-0612-061 | 1One | 46.3±5.6* 46.3 ± 5.6 * |
| 3-0023-002 | 1One | 16.4±6.216.4 ± 6.2 |
| 3-0103-010 | 1One | 30.8±7.0* 30.8 ± 7.0 * |
| 3-0363-036 | 1One | 9.2±3.59.2 ± 3.5 |
| 3-0413-041 | 1One | 8.1±3.38.1 ± 3.3 |
| 3-0463-046 | 1One | 44.7±5.7* 44.7 ± 5.7 * |
| 3-0473-047 | 1One | 49.8±7.0* 49.8 ± 7.0 * |
| 3-0483-048 | 1One | 6.0±4.66.0 ± 4.6 |
| 3-0493-049 | 1One | 31.6±4.2* 31.6 ± 4.2 * |
| 3-0533-053 | 1One | 14.3±4.114.3 ± 4.1 |
| 3-0563-056 | 1One | 72.5±3.3* 72.5 ± 3.3 * |
| 3-0613-061 | 1One | 25.0±2.9* 25.0 ± 2.9 * |
| 3-0653-065 | 1One | 57.0±2.9* 57.0 ± 2.9 * |
| 3-0733-073 | 1One | 77.4±3.9* 77.4 ± 3.9 * |
| 3-0863-086 | 1One | 15.2±5.315.2 ± 5.3 |
| 3-1003-100 | 1One | 20.3±2.020.3 ± 2.0 |
| 3-1233-123 | 1One | 19.3±3.019.3 ± 3.0 |
* P <0.05 compared to vehicle-treated control group. * P <0.05 compared to vehicle-treated control group.
상기 표 5에 나타낸 바와 같이, IL-6로 유도된 단핵구-장상피세포 부착에 대해 5-ASA(양성대조군)의 경우 1000μM 농도에서 1.7%의 억제율을 나타내어 화합물들의 시험 농도인 1μM의 1000배 높은 농도에서도 그 효과가 거의 나타나지 않은 반면, 2-056 화합물은 약 78%의 억제율을 보였고, 3-073 화합물은 약 77%의 억제율을 나타내었으며, 3-056 화합물은 약 73%의 억제율을 나타내어 매우 우수한 활성을 보였다. 화합물 2-036은 약 69%의 억제율을, 3-065는 약 57%의 억제율을, 3-047은 약 50%의 억제율을 보였으며, 화합물 2-041, 2-046, 3-046을 비롯한 대부분의 화합물이 대조 물질인 5-ASA에 비해서 우수한 억제 활성을 갖는 것으로 확인되었다. As shown in Table 5, the 5-ASA (positive control) showed 1.7% inhibition rate at 1000 μM concentration for IL-6-induced monocyte-intestinal epithelial cell adhesion, which was 1000 times higher than the test concentration of 1 μM. While almost no effect was found at the concentration, the 2-056 compound showed about 78% inhibition, the 3-073 compound showed about 77% inhibition, and the 3-056 compound showed about 73% inhibition. It showed excellent activity. Compound 2-036 exhibited an inhibition rate of about 69%, 3-065 exhibited an inhibition rate of about 57%, and 3-047 exhibited an inhibition rate of about 50%, including compounds 2-041, 2-046, and 3-046. It was found that most of the compounds had superior inhibitory activity compared to the 5-ASA control.
[시험예 2][Test Example 2]
TNBS로 유도한 염증성 장질환 동물 모델에서 화합물의 경구 투여 in vivo 효능 시험In vivo Efficacy Testing of Compounds in TNBS-Induced Inflammatory Bowel Disease Animal Models
<시험 방법><Test method>
동물은 7-8주령된 Sprague Dawley Rat을 오리엔트바이오(Korea)로부터 구입하여 3일간 일반 고형 사료로 안정화 시킨 후 실험에 이용하였다. 실험 기간 중 사료와 물을 자유로이 공급하였고, 사육실의 온도는 25±1℃, 상대습도는 50±10%로 유지시켰다. 점등 관리는 자동조명조절기에 의해 12시간 명암주기(light-dark cycle)로 조절하였다. 실험군은 각 군당 6마리로 하여 평균체중이 180±10 g이 되도록 난괴법 (randomized block design)에 의하여 5군(대조군, TNBS 단독 투여군, TNBS+설파살라진 300mg/kg 투여군, TNBS+시험약물 1mg/kg 투여군)으로 나누어 실험하였다.Animals were purchased for 7-8 weeks old Sprague Dawley Rat from Orient Bio (Korea) and stabilized with general solid feed for 3 days before being used for the experiment. Feed and water were freely supplied during the experiment, and the temperature in the cage was maintained at 25 ± 1 ℃ and relative humidity at 50 ± 10%. Lighting control was controlled by a 12-hour light-dark cycle by an automatic light controller. The experimental group consisted of 6 animals in each group, and the average weight was 180 ± 10 g by the randomized block design (5 groups (control group, TNBS-only group, TNBS + sulfasalazine 300mg / kg group, TNBS + test drug group)) The experiment was divided by.
(1) TNBS 직장 투여 장염 유발(1) TNBS enteral administration induced enteritis
24시간 절식한 rat를 diethyl ether로 마취한 후, polyethylene catheter를 연결한 1 mL 주사기를 이용하여 colon의 lumen에 50v/v% ethanol로 희석한 5% TNBS(2,4,6-trinitrobenzene sulfonic acid)를 0.8 mL을 천천히 주입한 후, 항문으로 5% TNBS가 새어 나오는 것을 방지하기 위하여 rat를 거꾸로 세운 상태에서 60초 동안 정치시켰다. 대조군은 vehicle [50v/v% ethanol]만을 다른 군과 마찬가지 방법으로 주입하였다 (Thapa et al., 2008).Rats fasted for 24 hours were anesthetized with diethyl ether, and then 5% TNBS (2,4,6-trinitrobenzene sulfonic acid) diluted with 50v / v% ethanol in the colon lumen using a 1 mL syringe connected with polyethylene catheter. After slowly injecting 0.8 mL of the rats, the rats were left upside down for 60 seconds to prevent 5% TNBS from leaking into the anus. The control group was injected with vehicle [50v / v% ethanol] in the same manner as the other groups (Thapa et al., 2008).
(2) 약물 투여(2) drug administration
약물의 효과를 조사하기 위하여 TNBS를 처치한 다음날부터 5일 동안 약물을 매일 일정한 시간에 투여하였다.To investigate the effect of the drug, the drug was administered at a fixed time every day for 5 days from the day after TNBS treatment.
(3) 체중 관찰(3) weight observation
Digital mass meter를 이용하여 절식 단계부터 TNBS 투여 및 약물 투여 과정 동안 각 rat의 체중 변화를 관찰하였다.The weight change of each rat was observed from the fasting stage to the TNBS administration and drug administration using a digital mass meter.
(4) 장 무게 측정(4) sheet weight measurement
Rat의 대장을 적출하여 항문으로부터 5-6 cm 사이의 조직을 1 cm 길이로 잘라서 조직의 무게를 측정하였다. The large intestine of the rat was extracted, and tissues between 5-6 cm from the anus were cut into 1 cm lengths and the tissues were weighed.
<TNBS로 유도한 장염증에 대한 화합물의 경구 투여 효과>Effect of Oral Administration of Compounds on Intestinal Inflammation Induced by TNBS
In vitro 부착 억제 시험에서 활성이 우수한 2-036 화합물을 대상으로 in vivo 장염 억제 활성을 측정하여 하기 표 6에 나타내었다. In vitro enteritis inhibitory activity was measured in 2-036 compound having excellent activity in in vitro adhesion inhibition test, and is shown in Table 6 below.
| 화합물compound | 투여 용량Dosage | 장 무게 회복률(%)Intestinal weight recovery (%) | 체중 회복률(%)Weight recovery rate (%) |
| 설파살라진Sulfasalin | 300 mg/kg300 mg / kg | 6464 | 4949 |
| 2-0362-036 | 1 mg/kg1 mg / kg | 7979 | 5959 |
A. 체중의 변화A. Change in weight
체중 180-190 g인 rat에 5% TNBS를 이용하여 장내에 염증을 유발한 대장염 모델에서 TNBS 처리 전의 몸무게를 기준으로 5일 간 매일 일정시간에 몸무게의 변화를 관찰한 결과, vehicle 처리 대조군은 계속해서 몸무게가 증가함을 보이고 TNBS만 처리한 군은 계속하여 몸무게가 감소하며 5일째부터 몸무게가 약간 회복되었으나, 정상군과 비교했을 때 몸무게가 현저히 감소되었다. 양성대조군인 설파살라진 300mg/kg을 처리한 군은 몸무게가 서서히 회복되어 vehicle 처리 대조군에 비해서는 감소되었지만, TNBS 단독 투여군에 비해 현저히 몸무게가 증가하여 몸무게 회복률 49%를 나타냈다. 화합물 2-036은 설파살라진 투여용량의 1/300인 1mg/kg만을 투여하였음에도 59%의 체중 회복률을 나타내었다.In the colitis model that induced inflammation in the intestine with 5% TNBS in rats weighing 180-190 g, the weight change was observed at a certain time every day for 5 days based on the weight before TNBS treatment. The weight increased and the TNBS-treated group continued to lose weight and recovered slightly from day 5, but the weight was significantly reduced compared to the normal group. The group treated with 300 mg / kg of sulfasalazine, a positive control group, recovered slowly and decreased compared to the vehicle-treated control group, but significantly increased weight compared to the TNBS-only group, showing 49% recovery. Compound 2-036 showed a 59% body weight recovery even when only 1 mg / kg, which was 1/300 of the sulfasalazine dose.
B. 형태학적 관찰B. Morphological Observations
5일 간의 약물투여가 끝난 후에 대장을 적출하여 육안으로 살펴 본 결과, TNBS를 처리한 rat의 대장은 대조군에 비하여 부종과 충혈이 관찰되었으며, 충수돌기의 부종과 울혈 및 장조직의 유착현상이 나타났다. 양성대조군인 설파살라진 300mg/kg을 처리한 군에서는 육안적 증상과 다른 기관들 사이의 유착이나 대장의 충혈도 현저히 억제되었으며, 화합물 2-036을 1mg/kg의 용량으로 투여한 그룹의 경우는 설파살라진 300mg/kg을 처리한 그룹보다 증상이 더욱 개선되었다.After 5 days of drug administration, the colon was extracted and visually examined. The colon of TNBS-treated rats showed edema and hyperemia compared to the control group, and edema of the appendix and congestion and adhesion of intestinal tissue were observed. . The group treated with 300 mg / kg of sulfasalazine, a positive control group, also significantly suppressed gross symptoms and adhesions between the other organs and colonic congestion, and sulfasalazine 300 mg in the group administered Compound 2-036 at a dose of 1 mg / kg. Symptoms improved more than the group treated with / kg.
C. 장 무게 측정C. Intestinal Weight Measurement
Rat의 대장을 적출하여 항문으로부터 5-6 cm 사이의 조직 무게를 측정한 결과, vehicle-처리 대조군에 비해 TNBS 단독 처리군의 경우 부종이 있는 장의 무게가 유의적으로 증가하였으며, 양성대조군인 설파살라진 300mg/kg을 처리한 군에서는 장조직 무게 회복률 64%를 나타냈다. 화합물 2-036은 설파살라진의 1/300의 용량인 1mg/kg만을 투여하였음에도 79%의 우수한 장 무게 회복률을 나타내었다. When the colon of the rat was harvested and the tissue weight was measured between 5-6 cm from the anus, the weight of the intestine with edema was significantly increased in the TNBS-treated group compared to the vehicle-treated control group, and 300 mg of sulfasalazine, a positive control group. In the group treated with / kg, the intestinal tissue recovery was 64%. Compound 2-036 showed an excellent intestinal weight recovery of 79% even when administered only 1 mg / kg, a dose of 1/300 of sulfasalazine.
[시험예 3][Test Example 3]
염증 T 세포 분화 억제 활성 시험 (in vitro)Inflammatory T cell differentiation inhibitory activity test (in vitro)
B57BL/6 마우스의 비장(spleen)으로부터 CD4 microbead를 이용하여 MACS라는 자석 장비를 통해 CD4 T 세포를 분리했다. CD4 T 세포를 자극하기 위해 CD3 항체(1mg/mL)를 세포를 키울 플레이트에 코팅 한 후, CD4 T 세포를 넣고 다른 자극제인 CD28 항체(1mg/mL)를 넣었다.CD4 T cells were isolated from a spleen of B57BL / 6 mice using a magnetic instrument called MACS. In order to stimulate CD4 T cells, CD3 antibody (1mg / mL) was coated on the plate to grow the cells. Then, CD4 T cells were added and other stimulant CD28 antibody (1mg / mL) was added.
T 세포에서 염증세포는 여러 종류가 있는데 가장 대표적인 염증 세포는 Th1, Th2 또는 Th17 세포이다. 이들 염증 세포는 분화 방법이 다르다. Th1 세포를 만들기 위하여, RPMI-1640 배지에 10% FBS와 1% penicillin/streptomycin (PS)를 포함시킨 후, 분리한 CD4 T 세포를 37℃, 5% CO2 조건 하에 배양하였다. 배양할 때 세포자극제인 CD3 항체와 CD28 항체를 앞에서와 같이 넣어주었다. Th1 세포로 분화시키기 위해 사이토카인으로 IL-12(10 ng/mL)을 넣어주고 4일 동안 키웠다. 이 때 약물의 억제 능력을 확인하기 위해 화합물 2-036(1μM) (처리군)을 포함시켰다.There are several types of inflammatory cells in T cells, the most representative of which are Th1, Th2 or Th17 cells. These inflammatory cells differ in their methods of differentiation. To make Th1 cells, 10% FBS and 1% penicillin / streptomycin (PS) were included in RPMI-1640 medium, and the isolated CD4 T cells were incubated under 37 ° C. and 5% CO 2 conditions. When cultured, the cell stimulant CD3 antibody and CD28 antibody were added as before. To differentiate into Th1 cells, IL-12 (10 ng / mL) was added as a cytokine and grown for 4 days. At this time, compound 2-036 (1 μM) (treatment group) was included to confirm the inhibitory ability of the drug.
대조군으로 류마티스 관절염 질환의 T 세포 억제제인 토파시티닙(tofacitinib, 1μM)과 트리암시놀론(triamcinolone, 1μM)을 사용하였다. 4일 후 세포를 PMA/ionomycin 및 golgi stop으로 4시간동안 재자극을 주고, PBS로 2번 세척하였다. 세척 후 Fix/Perm buffer를 이용하여 세포 표면에 구멍을 내고, PE-anti-IFN-γ 항체를 이용하여 유세포 분석기(flow cytometry)를 통해 IFN-γ를 분비하는 Th1 세포를 확인하였다.Tofacitinib (1 μM) and triamcinolone (1 μM), which are T cell inhibitors of rheumatoid arthritis, were used as controls. After 4 days, cells were restimulated with PMA / ionomycin and golgi stop for 4 hours and washed twice with PBS. After washing, the cell surface was punctured using Fix / Perm buffer, and Th1 cells secreting IFN-γ were identified through flow cytometry using PE-anti-IFN-γ antibody.
다른 염증 세포인 Th17 세포의 분화를 위하여, Th1 세포의 분화 방법과 동일하되, 사이토카인으로 IL-12 대신에 IL-6(10 ng/mL), TGF-β(1 ng/mL), anti-IFN-γ 항체(5 mg/mL) 및 anti-IL-4 항체(5 mg/mL)를 넣었다. 염증억제 세포인 조절T 세포(regulatory T cells)는 상기와 같이 동일한 방법으로 하되, 사이토카인으로 IL-2(10 ng/mL) 및 TGF-β(10 ng/mL)를 넣었다.For the differentiation of Th17 cells, which are other inflammatory cells, the same method as for the differentiation of Th1 cells, but instead of IL-12 as a cytokine, IL-6 (10 ng / mL), TGF-β (1 ng / mL), anti- IFN-γ antibody (5 mg / mL) and anti-IL-4 antibody (5 mg / mL) were added. Regulatory T cells, which are inflammation inhibitory cells, were prepared in the same manner as above, but IL-2 (10 ng / mL) and TGF-β (10 ng / mL) were added as cytokines.
본 발명에 따른 화합물의 염증세포 분화 억제 효과를 하기 표 7에 나타내었다. The inflammatory cell differentiation inhibitory effect of the compound according to the present invention is shown in Table 7 below.
| 화합물 (1μM)Compound (1 μM) |
Th 1 세포 분화억제 활성(%) |
Th 17 세포 분화억제 활성(%)Th 17 cell differentiation inhibitory activity (%) | |
| 2-0362-036 | 5555 | 5555 | |
| 2-0372-037 | 2222 | 2424 | |
| 2-0392-039 | 2727 | 1616 | |
| 2-0412-041 | 3737 | 2323 | |
| 2-0452-045 | 1010 | 3030 | |
| 2-0462-046 | 2626 | 1414 | |
| 2-0472-047 | 5555 | -11-11 | |
| 2-0482-048 | 2626 | 1313 | |
| 2-0492-049 | 3838 | 2222 | |
| 2-0532-053 | 1818 | 3737 | |
| 2-0562-056 | 2828 | 33 | |
| 2-0612-061 | 88 | 99 | |
| 3-0023-002 | 2323 | 66 | |
| 3-0103-010 | 4040 | 3737 | |
| 3-0363-036 | 4343 | 88 | |
| 3-0413-041 | 3333 | 3030 | |
| 3-0463-046 | 2424 | 1818 | |
| 3-0473-047 | 3737 | 1616 | |
| 3-0483-048 | 2727 | 1010 | |
| 3-0493-049 | 3434 | 1919 | |
| 3-0533-053 | -3-3 | 66 | |
| 3-0563-056 | 88 | -41-41 | |
| 3-0613-061 | -16-16 | 1515 | |
| 3-0653-065 | 1414 | 2020 | |
|
3-0733- | 1One | 2020 | |
| 3-0863-086 | 2222 | 1111 | |
| 3-1003-100 | 2323 | 2323 | |
| 3-1233-123 | 2525 | 1919 |
표 7에 나타낸 바와 같이, 본 발명 화합물의 Th1 및 Th17 세포의 in vitro 분화 억제 정도를 조사한 결과, 2-036 화합물이 Th1 및 Th17 세포 모두에서 50% 이상의 분화 억제율을 나타내었다. 그리고 화합물 2-036은 T 세포 분화 자극 조건에서 억제율이 높으면서 세포독성을 보이지 않는 화합물이었다.As shown in Table 7, when the degree of inhibition of differentiation of Th1 and Th17 cells of the compound of the present invention was examined, 2-036 compound showed a inhibition rate of 50% or more in both Th1 and Th17 cells. Compound 2-036 was a compound having high inhibition rate and no cytotoxicity under T cell differentiation stimulation conditions.
Th1 세포 분화 유도는 사이토카인 처리 3일 후 유세포분석기를 통하여 측정하여 도 1에 나타내었다. 사이토카인에 의한 Th1 분화 정도는 30% 정도 이루어졌다.Induction of Th1 cell differentiation was measured by flow cytometry 3 days after cytokine treatment and is shown in FIG. 1. The degree of Th1 differentiation by cytokines was about 30%.
화합물의 투여에 의한 사이토카인-유도성 Th1 세포 분화 정도는 화합물 2-036의 경우 13% 이하로 나타났으며, 양성대조군으로 사용한 토파시티닙과 트리암시놀론은 8% 정도로 나타났다(도 1A).The degree of cytokine-induced Th1 cell differentiation by the administration of the compound was 13% or less for Compound 2-036, and 8% of tofacitinib and triamcinolone used as positive controls (FIG. 1A).
또한 Th17 세포의 분화에 대해서도 화합물 2-036은 10% 이하의 분화를 보였고 토파시티닙은 5%, 트리암시놀론은 1% 정도의 분화가 보였다(도 1B).In addition, for the differentiation of Th17 cells, Compound 2-036 showed differentiation of 10% or less, tofacitinib, 5%, and triamcinolone about 1% (FIG. 1B).
하지만 토파시티닙과 트리암시놀론은 면역억제 세포(FoxP3+, Treg 세포)의 분화도 심각하게 억제시키는 반면, 화합물 2-036은 Th1와 Th17 세포 분화는 억제하고 면역억제 세포인 Treg 세포의 분화에는 영향을 나타내지 않았다(도 1C).However, tofacitinib and triamcinolone also significantly inhibited the differentiation of immunosuppressive cells (FoxP3 +, Treg cells), whereas Compound 2-036 inhibited the differentiation of Th1 and Th17 cells and did not affect the differentiation of Treg cells, which are immunosuppressive cells. (FIG. 1C).
따라서, 화합물 2-036은 염증 세포에만 특이적으로 반응하는 것을 확인할 수 있다. Thus, it can be seen that Compound 2-036 specifically responds only to inflammatory cells.
[시험예 4][Test Example 4]
세포 독성 시험 (in vitro)Cytotoxicity test (in vitro)
세포를 염증세포 자극 조건에서 3일 동안 자극한 후 PI와 Annexin V를 염색하여 죽어가는 세포 또는 죽은 세포를 구별하여 세포 독성을 확인하였다.Cells were stimulated under inflammatory cell stimulation conditions for 3 days, and then stained with PI and Annexin V to identify dying cells or dead cells.
화합물 2-036의 세포독성을 정확하게 측정하기 위해 다양한 농도에서 독성 시험을 하고 그 결과를 도 2에 나타냈었다. 도 2의 세포 분열 그래프에서 나타나듯이, 화합물 2-036은 모든 농도에서 독성을 나타내지 않았다(도 2A).To accurately measure the cytotoxicity of the compound 2-036 was tested for toxicity at various concentrations and the results are shown in FIG. As shown in the cell division graph of FIG. 2, compound 2-036 did not show toxicity at all concentrations (FIG. 2A).
반면, 양성 대조 약물인 토파시티닙과 트리암시놀론은 낮은 약물처리농도에서부터 세포 독성이 유의적으로 심하게 나타났다(도 2B 및 2C).On the other hand, the positive control drugs tofacitinib and triamcinolone showed significant severe cytotoxicity from low drug concentrations (FIGS. 2B and 2C).
이들 대조 약물은 세포 독성이 나타나서, 분화 억제와 독성 모두에 영향을 미쳐 약물의 특이성이 떨어진다. 반면, 화합물 2-036은 오직 T세포 분화에만 억제 능력을 보이고, T세포 자체에 대한 독성은 보이지 T세포 분화에 대한 특이성이 매우 높은 약물로 평가된다. 따라서, 화합물 2-036은 T세포의 염증 분화에만 관여하기 때문에 다른 세포에 대한 영향이 적고 부작용이 작을 뿐만 아니라 효과를 극대화할 수 있는 가능성을 보여준다. These control drugs are cytotoxic, affecting both differentiation inhibition and toxicity, resulting in poor drug specificity. On the other hand, Compound 2-036 shows only inhibitory ability to T cell differentiation and no toxicity to T cell itself, and is evaluated as a drug having a very high specificity for T cell differentiation. Therefore, compound 2-036 is only involved in inflammatory differentiation of T cells, thus showing the possibility of minimizing the effects on other cells and minimizing side effects and maximizing the effect.
[시험예 5][Test Example 5]
다발성 경화증 동물 모델(EAE)에서 화합물 2-036의 in vivo 효능 시험In vivo Efficacy Test of Compound 2-036 in Multiple Sclerosis Animal Model (EAE)
동물은 8~12주령 된 B57BL/6 마우스를 사용하였으며, 사육 기간 동안 사료와 물을 자유로이 공급하였고, 사욱실의 온도는 25±1℃, 상대습도는 50±10%로 유지시켰다. 점등 관리는 자동조명조절기의 의해 12시간 명암주기로 조절하였다. 실험군은 각 군당 5마리로 하여 난괴법(randomized block design)에 의하여 2군(대조군, 약물투여군)으로 나누어 실험하였다.B57BL / 6 mice, 8-12 weeks old, were used to feed and water freely during the breeding period, and the temperature of the room was 25 ± 1 ℃ and relative humidity was 50 ± 10%. Lighting control was controlled by a 12 hour contrast cycle by an automatic light controller. The experimental group was divided into 2 groups (control group, drug administration group) by randomized block design with 5 animals in each group.
(1) Experimental autoimmune encephalomyelitis (EAE) 유발(1) Induce experimentalal autoimmune encephalomyelitis (EAE)
피하주사로 6 mg/mL의 합성 펩티드 myelin oligodendrocyte glycoprotein(MOG, 신경계 항원, MEVGWYRSPFSRVVHLYRNGK; 펩티드 시퀀스)를 complete fluid adjuvant(CFA, 10 mg/ml heat-killed H37Ra, strain of Mycobacterium tuberculosis 포함된 면역증강제)와 섞여 주입하였다. 이틀 후 pertussis toxin 250 ng을 복강 주사로 주입하였다.Subcutaneous injection of 6 mg / mL synthetic peptide myelin oligodendrocyte glycoprotein (MOG, nervous system antigen, MEVGWYRSPFSRVVHLYRNGK; peptide sequence) with complete fluid adjuvant (CFA, 10 mg / ml heat-killed H37Ra, strain of Mycobacterium tuberculosis) It was mixed and injected. Two days later, 250 ng of pertussis toxin was injected by intraperitoneal injection.
대조군은 PBS를 격일로 복강으로 주입하였고 약물투여군은 화합물 2-036을 3mg/kg 양으로 격일로 복강에 주입하였다. 질환의 심각성 정도는 0에서 5까지로 나누었다.The control group was intraperitoneally injected with PBS every other day, and the drug administration group was intraperitoneally injected with compound 2-036 every other day in an amount of 3 mg / kg. The severity of the disease was divided from 0 to 5.
0: 아무 증상이 없음0: no symptoms
1: 꼬리가 축쳐진 상태1: tail is damp
2: 말단의 일부 마비 증세2: terminal paralysis
3: 양측 하지 마비3: bilateral paralysis
4: 네 다리 모두 마비4: paralyzed all four legs
5: 죽음5: death
보통 항원 주입 후 3주까지 관찰하며 그 이후에는 동물모델에서는 자생적으로 회복된다.It is usually observed up to 3 weeks after antigen injection, after which the animal recovers spontaneously.
(2) 면역 세포 분석을 통한 약물 억제 효과 확인(2) Confirmation of drug inhibitory effect through immune cell analysis
항원 주입 후 2주 뒤 마우스에서 비장(spleen), 림프절(LN), 뇌와 척수(CNS)를 적출하여 면역 세포 수를 계산하고, Th1 및 Th17 세포의 분포를 확인하기 위해 각 기관에서 얻은 면역 세포를 PMA/Ionomycin 및 golgi stop를 이용하여 자극하였다.Two weeks after antigen injection, the spleen, lymph node (LN), brain and spinal cord (CNS) were extracted from the mouse to calculate the number of immune cells, and the immune cells obtained from each organ to check the distribution of Th1 and Th17 cells. Was stimulated with PMA / Ionomycin and golgi stop.
그 후, Th1 세포를 확인할 수 있는 anti-IFN-γ 항체와 Th17 세포를 확인할 수 있는 anti-IL-17 항체를 사용하여 염색한 후 유세포 분석기(flow cytometry)를 이용하여 전체 면역 세포 중에서 염증 세포의 비중을 확인하였다. 그 결과를 도 3에 나타내었다.Thereafter, the cells were stained with an anti-IFN-γ antibody capable of identifying Th1 cells and an anti-IL-17 antibody capable of identifying Th17 cells, followed by flow cytometry. Specific gravity was confirmed. The results are shown in FIG.
도 3의 그래프를 보면, 항원 주입 후 일주일이 지나면 마우스의 행동패턴이 비정상적인 모습을 보이기 시작하는데, 대조군에서는 일주일부터 움직임이 적고 10일부터 꼬리가 축 쳐지면서 꼬리 움직임이 확연히 줄며, 14주가 되면 꼬리에 마비가 발생하고, 2-3일 뒤에는 하반신 마비가 시작되었다.In the graph of Figure 3, a week after the antigen injection, the behavioral pattern of the mouse begins to show an abnormal appearance, in the control group less movement from a week and the tail is drooping from the 10th, the tail movement is significantly reduced, the tail at 14 weeks Paralysis occurred in the lower body, followed by paraplegia after 2-3 days.
그러나, 격일로 화합물 2-036을 투여한 군에서는 행동 패턴의 이상이 10일 이후에 비로소 나타나면서 대조군보다 증상이 2-3 일 늦게 나타났다. 또한, 이상 증세 정도도 약하여 꼬리 마비까지만 나타나고 하반신 마비 증세를 볼 수 없었다(도 3A).However, in the group treated with Compound 2-036 every other day, symptoms of behavioral patterns appeared only after 10 days, and symptoms appeared 2-3 days later than the control group. In addition, the degree of abnormality was also weak, until only tail paralysis and paraplegia could not be seen (Fig. 3A).
조직 검사를 하여 확인해본 결과 척수에 면역세포 침입이 약물을 통해 줄어들어 있으며 (도 3D, 3E) 신경을 둘러쌓고 있는 미엘린수초도 질환유도 그룹에서는 많이 파괴되어 있는 반면 2-036 약물처리 그룹에서는 미엘린수초가 잘 보전되어 있었다. (도 3D, 3 F)The results of histological examination confirmed that the invasion of immune cells into the spinal cord was reduced through drugs (FIGS. 3D and 3E), and myelin sheaths surrounding the nerves were much destroyed in disease-induced groups, while myelin sheaths in the 2-036 drug treatment group. Was well preserved. (Figure 3D, 3F)
따라서 in vivo에서 본 발명에 따른 약물의 질환 억제 효과가 좋은 것을 확인할 수 있었다.Therefore, it was confirmed that the disease inhibiting effect of the drug according to the present invention is good in vivo.
비장, 림프구, 뇌 및 척수에서 면역 세포를 분리하여 분석한 결과에서도 전체 면역세포 수 자체가 약물 투여군에서 모두 줄어들어 있었고(도 3B, 3C), 염증 세포인 Th1 및 Th17 세포 분석 결과도 비장, 림프구, 뇌와 척수에서 유의적으로 줄어들어 있는 것을 확인하였다(도 3G, 3H).As a result of separating and analyzing immune cells in the spleen, lymphocytes, brain and spinal cord, the total immune cell number itself was reduced in the drug administration group (FIGS. 3B and 3C), and the inflammatory cells, Th1 and Th17 cells, were also analyzed in the spleen, lymphocyte, It was confirmed that significantly reduced in the brain and spinal cord (Fig. 3G, 3H).
따라서 화합물 2-036이 in vivo 에서도 T 세포의 염증 분화 억제 능력이 우수하고 EAE 질환 억제 효과도 우수함을 확인하였다.Therefore, it was confirmed that Compound 2-036 was excellent in inhibiting inflammatory differentiation of T cells and also in inhibiting EAE disease in vivo.
[시험예 6][Test Example 6]
DSS로 유도한 염증성 장질환 동물 모델에서 화합물의 경구 투여 in vivo 효능 시험In vivo Efficacy Testing of Compounds in DSS-induced Inflammatory Bowel Disease Animal Models
<시험방법><Test method>
동물은 8~12주령 된 B57BL/6 마우스를 사용하였으며, 사육 기간 동안 사료와 물을 자유로이 공급하였고, 사육실의 온도는 25±1℃, 상대습도는 50±10%로 유지시켰다. 점등 관리는 자동조명조절기의 의해 12시간 명암주기로 조절하였다. 실험군은 각 군당 5마리로 하여 난괴법(randomized block design)에 의하여 3군(대조군, DSS 단독투여군, DSS+약물투여군)으로 나누어 실험하였다.Animals were 8-12 weeks old B57BL / 6 mice, freely supplied with feed and water during the breeding period, and maintained at 25 ± 1 ℃ and 50 ± 10% relative humidity. Lighting control was controlled by a 12 hour contrast cycle by an automatic light controller. The experimental group was divided into three groups (control group, DSS alone administration group, DSS + drug administration group) by randomized block design with 5 animals in each group.
(1) DSS 구강 투여 장염 유발(1) DSS oral administration of enteritis
DSS를 2.5%농도로 물에 녹여 그 물을 6일동안 섭취하게 하여 장염을 유발하였다.Enteritis was induced by dissolving DSS in water at 2.5% concentration for 6 days.
(2) 체중 관찰(2) weight observation
Digital mass meter를 이용하여 절식 단계부터 DSS 투여 및 약물 투여 과정 동안 각 마우스의 체중 변화를 관찰하였다.Digital mass meter was used to observe the weight change of each mouse during fasting and DSS and drug administration.
(3) 장 길이 측정(3) sheet length measurement
마우스의 대장을 적출하여 조직의 길이를 측정하였다. The colon of the mouse was removed and the length of the tissue was measured.
<DSS로 유도한 장염증에 대한 화합물의 경구 투여 효과>Effect of Oral Administration of Compounds on Intestinal Inflammation Induced by DSS
In vitro 염증 T 세포 분화 억제 시험에서 활성이 우수한 2-036 화합물을 대상으로 in vivo 장염 억제 활성을 측정하여 효과를 확인하였다.In vitro inflammatory T cell differentiation inhibition test was confirmed by measuring the in vivo enteritis inhibitory activity against 2-036 compound with excellent activity.
A. 체중의 변화A. Change in weight
체중이 약 20g인 마우스에 2.5% DSS를 이용하여 장내에 염증을 유발한 대장염 모델에서 DSS 처리 전의 몸무게를 기준으로 10일 간 매일 일정시간에 몸무게의 변화를 관찰한 결과, 물만 제공한 대조군은 계속해서 몸무게가 증가함을 보이고 DSS를 처리한 군은 몸무게가 3일째부터 감소가 시작되어 계속 감소가 진행되었으며, 정상군과 비교했을 때 몸무게가 현저히 감소되었다. 반면, DSS와 2-036 약물을 투여한 그룹에서는 대조군에 비해 몸무게가 증가되지는 않았지만 몸무게 감소가 상당히 억제되는 것을 확인하였다 (도 4A).In a model of inflammation-induced colitis with 2.5% DSS in a mouse weighing about 20 g, the weight change was observed daily for 10 days based on the weight before DSS treatment. As the weight increased and the DSS-treated group began to decrease on the 3rd day, the weight continued to decrease, and the weight was significantly decreased compared to the normal group. On the other hand, in the group administered the DSS and 2-036 drug, it was confirmed that the weight loss is significantly suppressed compared to the control group (Fig. 4A).
B. 형태학적 관찰B. Morphological Observations
6일 간의 약물투여가 끝난 후에 4일이 지난 다음 대장을 적출하여 육안으로 살펴본 결과, DSS를 처리한 마우스의 대장은 대조군에 비하여 장의 길이가 현저히 줄어들어 있는 것을 확인하였고, DSS+2-036약물 처리군은 장의 길이가 회복되어 있었다(도 4B).After 6 days of drug administration, the colon was extracted and examined visually after 4 days, and the colon of DSS-treated mice was found to have significantly reduced intestinal length compared to the control group, and DSS + 2-036 drug treatment. The group had recovered its bowel length (FIG. 4B).
C. 면역세포 분석C. Immune Cell Analysis
또한, 림프절에서 면역 세포를 분리하여 분석한 결과에서도 전체 면역세포 수 자체가 약물 투여군에서 모두 줄어들어 있었고(도 4C), 염증 세포인 Th1 및 Th17 세포 분석 결과도 유의적으로 줄어들어 있는 것을 확인하였다(도 4D). In addition, as a result of separating and analyzing immune cells from lymph nodes, the total number of immune cells itself was reduced in the drug administration group (FIG. 4C), and the results of inflammatory cells, Th1 and Th17 cells, were also significantly reduced (FIG. 4C). 4D).
Claims (9)
- 하기 화학식 1의 화합물, 또는 이의 약제학적으로 허용 가능한 염:A compound of Formula 1, or a pharmaceutically acceptable salt thereof:[화학식 1][Formula 1]
상기 식에서,WhereZ는 O 또는 S이고,Z is O or S,R은 수소; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 헤테로아릴알킬; 아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 벤질기로 치환되거나 치환되지 않는 피롤리돈; 알킬벤젠설포네이트; 벤조다이옥솔(benzodioxol); 또는 2,3-디하이드로-1,4-벤조다이옥신(2,3-dihydro-1,4-benzodioxin)이고,R is hydrogen; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),상기 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 및 헤테로아릴알킬은The alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl히드록시; 보로네이트; 할로겐; 할로알킬; 할로알콕시; 할로아릴; 아미노; 알킬아미노; 디알킬아미노; 아실아미노; 아릴카르보닐아미노; 술폰아미드; 니트로; 시아노; 카르보닐; 아실; 아미노카르보닐; 알킬아미노카르보닐; 디알킬아미노카르보닐; 히드라지드; 카르복실; 알콕시카르보닐; 아릴술포닐; 알킬; 알콕시; 알케닐; 알키닐; 시클로알킬; 아릴; 아릴알킬; 디아릴알킬; 아릴알콕시; 아세트아미드; 벤즈아미드; 알킬 및 할로겐으로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않은 아릴옥시; 알킬로 치환되거나 치환되지 않은 헤테로시클릭; 및 알킬로 치환되거나 치환되지 않은 헤테로아릴로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않으며,Hydroxy; Boronate; halogen; Haloalkyl; Haloalkoxy; Haloaryl; Amino; Alkylamino; Dialkylamino; Acylamino; Arylcarbonylamino; Sulfonamides; Nitro; Cyano; Carbonyl; Acyl; Aminocarbonyl; Alkylaminocarbonyl; Dialkylaminocarbonyl; Hydrazide; Carboxyl; Alkoxycarbonyl; Arylsulfonyl; Alkyl; Alkoxy; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Arylalkyl; Diarylalkyl; Arylalkoxy; Acetamide; Benzamide; Aryloxy unsubstituted or substituted with one or more groups selected from the group consisting of alkyl and halogen; Heterocyclic, optionally substituted with alkyl; And at least one group selected from the group consisting of heteroaryl unsubstituted or substituted with alkyl,여기서, 알킬은 C1-30 알킬이고,Wherein alkyl is C 1-30 alkyl,알콕시는 C1-30 알콕시이고,Alkoxy is C 1-30 alkoxy,알케닐은 C2-30 알케닐이고,Alkenyl is C 2-30 alkenyl,알키닐은 C2-30 알키닐이고,Alkynyl is C 2-30 alkynyl,시클로알킬은 C3-30 시클로알킬이고,Cycloalkyl is C 3-30 cycloalkyl,아릴은 C5-30 아릴이고,Aryl is C 5-30 aryl,헤테로시클릭은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로시클릭이고,Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,헤테로아릴은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로아릴임. Heteroaryl is heteroaryl having 3 to 30 ring atoms, each of which one or more ring carbons is replaced with a heteroatom selected from B, N, O, S. - 하기 화학식 1의 화합물, 또는 이의 약제학적으로 허용 가능한 염:A compound of Formula 1, or a pharmaceutically acceptable salt thereof:[화학식 1][Formula 1]
상기 식에서,WhereZ는 O 또는 S이고,Z is O or S,R은 수소; 알킬; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 헤테로아릴알킬; 아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 헤테로아릴기가 시클로알킬기에 융합되어 있는 융합고리기; 벤질기로 치환되거나 치환되지 않는 피롤리돈; 알킬벤젠설포네이트; 벤조다이옥솔(benzodioxol); 또는 2,3-디하이드로-1,4-벤조다이옥신(2,3-dihydro-1,4-benzodioxin)이고,R is hydrogen; Alkyl; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; Heteroarylalkyl; A fused ring group in which an aryl group is fused to a cycloalkyl group; A fused ring group in which a heteroaryl group is fused to a cycloalkyl group; Pyrrolidone substituted or unsubstituted with a benzyl group; Alkylbenzenesulfonate; Benzodioxol; Or 2,3-dihydro-1,4-benzodioxin (2,3-dihydro-1,4-benzodioxin),상기 알킬; 알케닐; 알키닐; 시클로알킬; 아릴; 헤테로시클릭; 헤테로아릴; 아릴알킬; 헤테로시클릭알킬; 및 헤테로아릴알킬은The alkyl; Alkenyl; Alkynyl; Cycloalkyl; Aryl; Heterocyclic; Heteroaryl; Arylalkyl; Heterocyclic alkyl; And heteroarylalkyl히드록시; 보로네이트; 할로겐; 할로알킬; 할로알콕시; 할로아릴; 아미노; 디알킬아미노; 아실아미노; 아릴카르보닐아미노; 술폰아미드; 니트로; 시아노; 히드라지드; 카르복실; 알콕시카르보닐; 아릴술포닐; 알킬; 알콕시; 아릴; 디아릴알킬; 아릴알콕시; 아세트아미드; 벤즈아미드; 알킬 및 할로겐으로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않은 아릴옥시; 알킬로 치환되거나 치환되지 않은 헤테로시클릭; 및 알킬로 치환되거나 치환되지 않은 헤테로아릴로 이루어진 군으로부터 선택된 하나 이상의 기로 치환되거나 치환되지 않으며,Hydroxy; Boronate; halogen; Haloalkyl; Haloalkoxy; Haloaryl; Amino; Dialkylamino; Acylamino; Arylcarbonylamino; Sulfonamides; Nitro; Cyano; Hydrazide; Carboxyl; Alkoxycarbonyl; Arylsulfonyl; Alkyl; Alkoxy; Aryl; Diarylalkyl; Arylalkoxy; Acetamide; Benzamide; Aryloxy unsubstituted or substituted with one or more groups selected from the group consisting of alkyl and halogen; Heterocyclic, optionally substituted with alkyl; And at least one group selected from the group consisting of heteroaryl unsubstituted or substituted with alkyl,여기서, 알킬은 C1-18 알킬이고,Wherein alkyl is C 1-18 alkyl,알콕시는 C1-18 알콕시이고,Alkoxy is C 1-18 alkoxy,알케닐은 C2-18 알케닐이고,Alkenyl is C 2-18 alkenyl,알키닐은 C2-18 알키닐이고,Alkynyl is C 2-18 alkynyl,시클로알킬은 C3-18 시클로알킬이고,Cycloalkyl is C 3-18 cycloalkyl,아릴은 C5-18 아릴이고,Aryl is C 5-18 aryl,헤테로시클릭은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로시클릭이고,Heterocyclic is a heterocyclic having from 3 to 30 ring atoms in which one or more ring carbons are each replaced with a heteroatom selected from B, N, O, S,헤테로아릴은 하나 이상의 고리 탄소가 각각 B, N, O, S로부터 선택된 헤테로원자로 대체되는 3 내지 30개의 고리 원자를 갖는 헤테로아릴임. Heteroaryl is heteroaryl having 3 to 30 ring atoms, each of which one or more ring carbons is replaced with a heteroatom selected from B, N, O, S. - 하기 화학식 2-036, 2-037, 2-039, 2-041, 2-045, 2-046, 2-047, 2-048, 2-049, 2-053, 2-056, 2-061, 3-002, 3-010, 3-036, 3-041, 3-046, 3-047, 3-048, 3-049, 3-053, 3-056, 3-061, 3-065, 3-073, 3-086, 3-100 또는 3-123로 표시되는 화합물, 또는 이의 약제학적으로 허용 가능한 염.Formulas 2-036, 2-037, 2-039, 2-041, 2-045, 2-046, 2-047, 2-048, 2-049, 2-053, 2-056, 2-061, 3-002, 3-010, 3-036, 3-041, 3-046, 3-047, 3-048, 3-049, 3-053, 3-056, 3-061, 3-065, 3- A compound represented by 073, 3-086, 3-100, or 3-123, or a pharmaceutically acceptable salt thereof.[화학식 2-036][Formula 2-036]
[화학식 2-037][Formula 2-037]
[화학식 2-039][Formula 2-039]
[화학식 2-041][Formula 2-041]
[화학식 2-045][Formula 2-045]
[화학식 2-046][Formula 2-046]
[화학식 2-047][Formula 2-047]
[화학식 2-048][Formula 2-048]
[화학식 2-049][Formula 2-049]
[화학식 2-053][Formula 2-053]
[화학식 2-056][Formula 2-056]
[화학식 2-061][Formula 2-061]
[화학식 3-002][Formula 3-002]
[화학식 3-010][Formula 3-010]
[화학식 3-036][Formula 3-036]
[화학식 3-041][Formula 3-041]
[화학식 3-046][Formula 3-046]
[화학식 3-047][Formula 3-047]
[화학식 3-048][Formula 3-048]
[화학식 3-049][Formula 3-049]
[화학식 3-053][Formula 3-053]
[화학식 3-056][Formula 3-056]
[화학식 3-061][Formula 3-061]
[화학식 3-065][Formula 3-065]
[화학식 3-073][Formula 3-073]
[화학식 3-086][Formula 3-086]
[화학식 3-100][Formula 3-100]
[화학식 3-123][Formula 3-123]
- 하기 화학식 2-036로 표시되는 화합물, 또는 이의 약제학적으로 허용 가능한 염. A compound represented by the following Chemical Formula 2-036, or a pharmaceutically acceptable salt thereof.[화학식 2-036][Formula 2-036]
- 하기 화학식 2-056로 표시되는 화합물, 또는 이의 약제학적으로 허용 가능한 염. A compound represented by the following Chemical Formula 2-056, or a pharmaceutically acceptable salt thereof.[화학식 2-056][Formula 2-056]
- 하기 화학식 3-056로 표시되는 화합물, 또는 이의 약제학적으로 허용 가능한 염. A compound represented by the following Chemical Formula 3-056, or a pharmaceutically acceptable salt thereof.[화학식 3-056][Formula 3-056]
- 하기 화학식 3-073로 표시되는 화합물, 또는 이의 약제학적으로 허용 가능한 염. A compound represented by the following Chemical Formula 3-073, or a pharmaceutically acceptable salt thereof.[화학식 2-073][Formula 2-073]
- 제1항의 화합물을 포함하는 염증성 장질환 예방 또는 치료용 약학적 조성물. A pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising the compound of claim 1.
- 제1항의 화합물을 포함하는 자가면역 질환 예방 또는 치료용 약학적 조성물. A pharmaceutical composition for preventing or treating autoimmune disease, comprising the compound of claim 1.
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| KR20020027635A (en) * | 1999-09-10 | 2002-04-13 | 폴락 돈나 엘. | Tyrosine kinase inhibitors |
| US20080255203A1 (en) * | 2007-04-12 | 2008-10-16 | Abbott Laboratories | Heterocyclic compounds and their methods of use |
| WO2011069647A1 (en) * | 2009-12-10 | 2011-06-16 | Almirall, S.A. | New 2-aminothiadiazole derivatives |
| WO2016115463A1 (en) * | 2015-01-15 | 2016-07-21 | Whitehead Institute For Biomedical Research | Inhibitors of phosphoglycerate dehydrogenase (phgdh) and uses thereof |
| WO2017208032A1 (en) * | 2016-06-03 | 2017-12-07 | Karus Therapeutics Ltd | Combinations comprising histone deacetylase inhibitors |
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| KR101415742B1 (en) * | 2011-12-21 | 2014-07-04 | 영남대학교 산학협력단 | 6-aminopyridin-3-ol derivative or a pharmaceutically acceptable salt thereof and pharmaceutical composition for treating or preventing angiogenesis-related disease comprising the same |
| US10604487B2 (en) * | 2016-01-14 | 2020-03-31 | Research Cooperation Foundation Of Yeungnam University | Pyridinol derivative or pharmaceutically acceptable salt thereof, and pharmaceutical composition containing same as active ingredient |
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| KR20020027635A (en) * | 1999-09-10 | 2002-04-13 | 폴락 돈나 엘. | Tyrosine kinase inhibitors |
| US20080255203A1 (en) * | 2007-04-12 | 2008-10-16 | Abbott Laboratories | Heterocyclic compounds and their methods of use |
| WO2011069647A1 (en) * | 2009-12-10 | 2011-06-16 | Almirall, S.A. | New 2-aminothiadiazole derivatives |
| WO2016115463A1 (en) * | 2015-01-15 | 2016-07-21 | Whitehead Institute For Biomedical Research | Inhibitors of phosphoglycerate dehydrogenase (phgdh) and uses thereof |
| WO2017208032A1 (en) * | 2016-06-03 | 2017-12-07 | Karus Therapeutics Ltd | Combinations comprising histone deacetylase inhibitors |
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