WO2016032127A2 - Nouveaux composés présentant des activités antioxydantes et anti-inflammatoires en raison de la concurrence avec lps pour se lier à tlr4, et leur utilisation médicale - Google Patents

Nouveaux composés présentant des activités antioxydantes et anti-inflammatoires en raison de la concurrence avec lps pour se lier à tlr4, et leur utilisation médicale Download PDF

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WO2016032127A2
WO2016032127A2 PCT/KR2015/007655 KR2015007655W WO2016032127A2 WO 2016032127 A2 WO2016032127 A2 WO 2016032127A2 KR 2015007655 W KR2015007655 W KR 2015007655W WO 2016032127 A2 WO2016032127 A2 WO 2016032127A2
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dione
thioxodihydropyrimidine
trione
pyrimidine
compound
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PCT/KR2015/007655
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Korean (ko)
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WO2016032127A3 (fr
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정해영
문형룡
문경미
박윤정
김남득
박지영
김혜림
김철민
정지원
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부산대학교 산학협력단
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Priority claimed from KR1020150101640A external-priority patent/KR101686872B1/ko
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Publication of WO2016032127A2 publication Critical patent/WO2016032127A2/fr
Publication of WO2016032127A3 publication Critical patent/WO2016032127A3/fr

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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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/06—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D239/08—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms directly attached in position 2
    • C07D239/10—Oxygen or sulfur atoms

Definitions

  • the present invention relates to a novel compound and its medical use having the effect of antioxidant, anti-inflammatory and anti-aging to inhibit the activity of macrophages through competitive binding of lipopolysaccharide (LPS) to TLR4.
  • LPS lipopolysaccharide
  • Inflammatory reactions are stimulated by oxidative stress in the body, which in turn causes nucleic acid (DNA), the major protein in muscle, proteins, and fat, the major component of cell membrane, to deform or deactivate the cell.
  • DNA nucleic acid
  • Oxygen which is absorbed through normal respiration, enters the mitochondria, an energy-producing tissue within the cell, and burns, producing energy (ATP) necessary for maintaining body temperature, cell activity, and physical activity.
  • ATP energy
  • the fundamental defect of mitochondria is that oxygen is not completely burned, producing by-products, which are free radicals, which are free radicals.
  • life has a defense system that detoxifies, destroys or neutralizes toxic oxygen throughout the evolutionary process, which is responsible for many resistances in the body that are balanced by a redox balance. As aging is broken by the force of destructive free radicals and oxidative destruction stresses cells and tissues, the body function is reduced and at the same time inflammation is generated.
  • Macrophage is an immune cell that is distributed in all tissues of the animal and is responsible for the innate immune response in the human body, and is a white blood cell that plays an important role in the human immune system. Macrophages activated by external stimuli trigger inflammatory responses through the secretion of inflammatory mediators, causing inflammation of asthma, bronchitis, arthritis, multiple sclerosis, arteriosclerosis, stroke, degenerative brain diseases such as Alzheimer's disease or Parkinson's disease It causes diseases and the like and makes the disease worse.
  • Lipopolysaccharide one of the endotoxins, has been shown to contain pro- and lipases such as tumor necrosis factor-alpha (TNF- ⁇ ), interleukin-6 (IL-6), and interleukin-1 ⁇ (IL-1 ⁇ ) in Raw264.7 macrophage. It increases inflammatory cytokines and secretes inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2).
  • NO nitric oxide
  • PGE2 prostaglandin E2
  • COX-2 cyclooxygenase-2
  • NOS NO synthase
  • NOS an enzyme that produces NO, and cyclooxygenase (COX)
  • COX an enzyme that mediates the biosynthesis of various prostaglandins (PGs)
  • PGs prostaglandins
  • TLR2 / TLR4 mainly detects lipoproteins in bacteria's outer cell membrane / cell wall, and induces ROS (active oxygen species) causing oxidative stress.
  • ROS active oxygen species
  • TLR4 even when macrophages are stimulated by LPS stimulation, several inflammatory factors are generated by NF- ⁇ B activation, which induces pathologies of various diseases and is also involved in carcinogenesis.
  • NF- ⁇ B activation NF- ⁇ B activation
  • antioxidants having ONOO scavenging activity in order to protect cells from damage caused by cytotoxic substances such as free radicals and NO, ONOO, and many natural-derived or synthetic scavengers have been developed.
  • Antioxidants are used for the purpose of minimizing the loss of certain vitamins and essential amino acids or by delaying or preventing rancidity of maintenance products by reacting with free radicals rather than removing or absorbing oxygen.
  • Synthetic antioxidants commonly used in foods and pharmaceuticals include butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl galate (PG) and tertiary butyl hydro Quinone (TBHQ, Tertiary butyl hydroquinone), etc., but when administered to high concentrations in experimental animals are known to cause liver hypertrophy or carcinogenicity.
  • butylated hydroxytoluene has been shown to increase microsomal enzyme activity in the liver of laboratory animals through several studies, and controversy has been raised about the safety of these phenolic synthetic antioxidants. Usage is legally regulated. Accordingly, many studies have been made in anticipation of the development of natural antioxidants of safe and economical plant origin with high antioxidant effects.
  • antioxidants Recently, researches on antioxidants have been actively conducted.
  • tocopherols flavonoids, gossypol, sesamol (sesamol), oryzanol, vitamin C, and the like.
  • tocopherol and L-ascorbic acid are preferred as natural antioxidants.
  • tocopherol has high safety but low ability to stop oxidation reaction alone and is expensive. .
  • TLR4 antagonists that directly bind to TLR4 have been rarely reported. Among them, erytoran, made by Eisai of Japan in late 2009, binds to MD2 of TLR4 and suppresses its signaling mechanisms for sepsis. It is reported that it has entered the phase 3 clinical trial for the treatment of patients. However, due to its very high molecular weight, there is a difficulty in mass production of itoritoran.
  • An object of the present invention is to provide a novel compound having the effect of antioxidant, anti-inflammatory and anti-aging to inhibit the activity of macrophages through competitive binding of lipopolysaccharide (LPS) to TLR4.
  • LPS lipopolysaccharide
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating inflammatory diseases containing the novel compound as an active ingredient.
  • Another object of the present invention to provide a health functional food for preventing or improving inflammatory diseases, antioxidant or anti-aging containing a novel compound as an active ingredient.
  • R1 to R4 may be the same or different, respectively, any one of H, OH, Br or C1 to C4 alkoxy, and X is O or S.
  • the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases containing the compound as an active ingredient.
  • the present invention also provides a health functional food for preventing or improving inflammatory diseases containing the compound as an active ingredient.
  • the present invention also provides a dietary supplement for antioxidants containing the compound as an active ingredient.
  • the present invention also provides an anti-aging health functional food containing the compound as an active ingredient.
  • the present invention relates to a novel compound having antioxidant and anti-inflammatory activity through competitive binding with LPS to TLR4 and its medical use, wherein the compounds according to the present invention are small molecules that inhibit the activity of macrophages through competitive binding with LPS to TLR4.
  • the compounds according to the present invention are small molecules that inhibit the activity of macrophages through competitive binding with LPS to TLR4.
  • ROS reactive oxygen species
  • NF- ⁇ B transcription factor
  • this experiment was also verified in liver of C57BL / 6 mice. Therefore, the novel compounds can be used in various fields, such as drugs or health foods, which have little side effects and are safe in preventing and treating sepsis, metabolic diseases or cardiovascular diseases.
  • 1 is a graph of compounds targeting TLR4 inhibition and having inhibitory effect through screening of ROS and ONOO ⁇ .
  • FIG. 2 shows that the compound according to the present invention binds to the MD2 portion of TLR4 and competes with LPS through docking simulation.
  • 3 is a graph showing that compounds 42 and 50, which are novel compounds of the present invention, are stable to cytotoxicity at concentrations of 1 ⁇ M to 10 ⁇ M through cell viability assays.
  • Figure 4a is the increased oxidative stress in macrophages by LPS-a graph showing that (ROS, ONOO, NO) is present 42 times, 50 times reduced by the compound.
  • 4B is a graph showing that ROS, ONOO ⁇ increased by LPS in macrophages through fluorescent dyes is reduced by compounds 42 and 50.
  • FIG. 5 is a diagram showing that expression of COX-2 and iNOS, which are inflammation-inducing proteins, increased by LPS by treating compounds 42 and 50, is inhibited.
  • FIG. 6 is a diagram showing the expression and activity of NF- ⁇ B, a transcription factor of oxidative stress and inflammation-related proteins, wherein the activity of NF- ⁇ B phosphorylation at ser536 is 42 and 50 at the phosphorylation site of P65. It is a picture that is inhibited by burn compound.
  • Figure 7 shows the activity of NF- ⁇ B in macrophages, a picture of the effects of inhibiting the translocation of NF- ⁇ B by compounds 42, 50.
  • FIG. 8 shows the effect of translocation into the nucleus, using a luciferase reporter vector to which an NF- ⁇ B binding site is ligation, with activation of NF- ⁇ B increased by LPS. Is a concentration-dependent inhibition by compounds 42 and 50.
  • FIG. 9 is a graph showing the effect of inhibiting translocation of NF- ⁇ B by decreasing the concentration of IKKB activity and I ⁇ B ⁇ activity in a substance treated with compounds 42 and 50.
  • Figure 11 is, C57BL / 6 and ONOO ROS in liver and blood in order to determine the inhibitory effect of oxidative stress in the macrophages in the animal model were measured.
  • ROS and ONOO - were increased in both liver and blood in LPS group mice, and concentration-dependently decreased in compounds 42 and 50.
  • the present invention is a small molecular weight compound having scavenging ability of oxidative stress in Raw264.7 macrophage, which directly binds to TLR4 and increases tumor necrosis factor-alpha (TNF- ⁇ ), interleukin-6 through competitive binding with LPS. It inhibits pro-inflammatory cytokines such as (IL-6) and interleukin-1 ⁇ (IL-1 ⁇ ) and inhibits the secretion of inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2).
  • IL-6 pro-inflammatory cytokines
  • IL-1 ⁇ interleukin-1 ⁇
  • NO nitric oxide
  • PGE2 prostaglandin E2
  • NF- ⁇ B a transcription-inducing transcription factor
  • COX-2 cycloxygenase-2
  • NOS NO synthase
  • the present invention provides a compound represented by the following formula (1).
  • R1 to R4 may be the same or different, respectively, any one of H, OH, Br or C1 to C4 alkoxy, and X is O or S.
  • the compound is 5- (4-hydroxybenzyl) pyrimidine-2,4,6 (1H, 3H, 5H) -trione, 5- (3,4-dihydroxybenzyl) pyrimidine- 2,4,6 (1H, 3H, 5H) -trione, 5- (2,4-dihydroxybenzyl) pyrimidine-2,4,6 (1H, 3H, 5H) -trione, 5- ( 4-hydroxy-3-methoxybenzyl) pyrimidine-2,4,6 (1H, 3H, 5H) -trione, 5- (3-ethoxy-4-hydroxybenzyl) pyrimidine-2,4 , 6 (1H, 3H, 5H) -trione, 5- (3-hydroxy-4-methoxybenzyl) pyrimidine-2,4,6 (1H, 3H, 5H) -trione, 5- (4 -Methoxybenzyl) pyrimidine-2,4,6 (1H, 3H, 5H) -trione, 5- (4 -Methoxybenzyl) pyr
  • the compound is 5- (4-hydroxy-3-methoxybenzyl) -2-thioxodihydropyrimidine-4,6 (1H, 5H)-represented by the following general formula (2) or (3): Dione (compound 42) or 5- (4-hydroxy-3,5-dimethoxybenzyl) -2-thioxodihydropyrimidine-4,6 (1H, 5H) -dione (compound 50), It is not limited to this.
  • the present invention provides a composition for preventing or treating inflammatory diseases containing the compound as an active ingredient.
  • the compound may inhibit the activity of macrophages through competitive binding to lipopolysaccharide (LPS) to TLR4.
  • LPS lipopolysaccharide
  • the inflammatory disease may be asthma, bronchitis, sepsis, arthritis, hepatitis, rheumatoid arthritis, osteoarthritis, ulcerative colitis, myocarditis, multiple sclerosis or viral infection, but is not limited thereto.
  • the pharmaceutical composition may include a pharmaceutically acceptable carrier in addition to the active ingredient.
  • a pharmaceutically acceptable carrier are commonly used in pharmaceutical preparations, such as lactose, Dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzo Ate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but is not limited thereto.
  • the pharmaceutical composition may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like as an additive.
  • the method of administration of the pharmaceutical composition is determined depending on the degree of symptoms, usually topical administration is preferred.
  • the dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the degree of the disease, the age, sex, and weight of the patient, and may be administered once to several times daily.
  • the pharmaceutical composition may be administered to various mammals such as rats, mice, livestock, humans, and the like. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
  • compositions may be prepared in unit dose form or formulated using pharmaceutically acceptable carriers and / or excipients or may be prepared within a multi-dose container.
  • the formulation may be in the form of a solution, suspension, or emulsion, or may be in the form of an exercicide, extract, powder, granule, tablet, warning, lotion, ointment, or the like.
  • the present invention also provides a health functional food for preventing or improving inflammatory diseases containing the compound as an active ingredient.
  • the present invention also provides a dietary supplement for antioxidants containing the compound as an active ingredient.
  • the present invention also provides an anti-aging health functional food containing the compound as an active ingredient.
  • health functional food refers to a food having a bioregulatory function, such as prevention and treatment of diseases, biological defense, immunity, recovery of symptoms, aging inhibition, and should be harmless to the human body when taken in the long term.
  • the health functional food of the present invention can be prepared by a variety of methods known in the food science or pharmaceutical field, any food which can be ingested orally in combination with itself or a food-acceptable carrier, excipient, diluent, etc. It may also be prepared in form. Preferably in the form of beverages, pills, granules, tablets or capsules.
  • the health functional food of the present invention may further include ingredients that are conventionally added and food-acceptable at the time of food production.
  • Table 1 shows 5- (substituted benzylidene) pyrimidine-2,4,6 (1H, 3H, 5H) -trione analogue [5- (substituted benzylidene) pyrimidine-2,4,6 (1H, 3H, 5H) -trione analog] For explaining the substitution pattern of compound 1-12.
  • OMe represents methoxy and OEt represents an ethoxy group.
  • Table 2 shows 5- (substituted benzylidene) -2-thioxodihydropyrimidine-4,6 (1H, 5H) -dione analogue [5- (substituted benzylidene) -2-thioxodihydropyrimidine-4,6 (1H). , 5H) -dione analog] To explain the substitution pattern of compound 13-26.
  • OMe represents methoxy and OEt represents an ethoxy group.
  • Table 3 shows the 5- (substituted benzyl) pyrimidine-2,4,6 (1H, 3H, 5H) -trione analogue [5- (substituted benzyl) pyrimidine-2,4,6 (1H, 3H, 5H). ) -trione analog] is for explaining the substitution pattern of compound 27-38.
  • Table 4 shows 5- (substituted benzyl) -2-thioxodihydropyrimidine-4,6 (1H, 5H) -dione analogs [5- (substituted benzyl) -2-thioxodihydropyrimidine-4,6 (1 H) , 5 H) -dione analog] serve to explain the substitution pattern of the compounds 39-52.
  • Dione are suspended in (for compounds 13 ⁇ 26, 30 mg) in ethanol (5 mL) - 5- (substituted-benzylidene) -2-thioxo-dihydro-pyrimidine -4,6 (1 H, 5 H) Sodium borohydride (NaBH 4 , 3 equiv) was slowly added into the suspended solution at 0 ° C. and the mixed solution was stirred at room temperature for 10 minutes-2 hours. The solvent was evaporated under reduced pressure, water was added, and the pH was adjusted to 7 with 1N hydrochloric acid (HCl).
  • HCl 1N hydrochloric acid
  • the water was volatilized under reduced pressure, then ether / water (10-20: 1) or ethanol / water (10: 1, for compounds 41, 46 and 47) was added and the resulting solid was filtered and ether / water (10 ⁇ 20: 1) or washing with ethanol / water (10: 1, for compounds 41, 46 and 47) to afford the desired product.
  • Raw 264.7 macrophagy cells (rat prostatic endothelial cell line) were obtained from American Type Culture Collection (ATCC), Manassas, VA, USA, which cells were 2 mM L-glutamine, 100 mg / ml streptomycin, 2.5 mg / L amphotericin. B, and cultured using DMEM (Dulbecco's Modified Eagle Medium, Nissui, Tokyo, Japan) containing 5% inactivated fetal bovine serum (FBS). The cells were also maintained at 37 ° C. under the same conditions as a humid atmosphere containing 5% CO 2 and 95% air. And 5% FBS was not added was used as a serum-free medium (SFM, serum-free medium). Cell lines were maintained by subculture every two days in a 100 mm plastic flask (Corning Co., New York, USA).
  • SFM serum-free medium
  • the fat-soluble DCFDA is deacetylated to non-fluorescent DCFH by esterase or oxidative hydrolysis, and DCFH is oxidized by free radicals to become DCF (2 ', 7'-dichlorofluorescein) which shows strong fluorescence, excitation wavelength 485nm And it was measured by a fluorescence photometer (GENios, TECAN) at an emission wavelength of 530nm.
  • ROS is optionally generated by reacting with esterase using 50 ⁇ M of 3-morpholinosydnonimine hydrochloride (SIN-1).
  • ONOO - scavenging and production inhibitory activity of the 26 novel compounds was mediated by ONOO - according to the method of Kooy et al., Free Radic. Biol. Med. 16: 149-156,1994. It was measured and analyzed by fluorescence spectroscopy using an oxidation reaction of dihydroramine (dihidrorhodamine 123; hereinafter referred to as "DHR"). Non-fluorescent DHR 123 is oxidized by ONOO ⁇ to form fluorescent rhodamine 123.
  • DHR dihydroramine
  • the affinity was confirmed through docking simulation in order to confirm whether the two novel compounds selected in FIG. 1 have an inhibitory effect and influence on the TLR4 receptor which directly affects the NF- ⁇ B activation pathway.
  • No. 42 and No. 50 were found to bind directly to the MD2 portion of the TLR4 complex, and as a result of confirming the intimacy of each compound, Compound No. 42 had two more methoxy groups than Compound No. 42. Since the affinity was higher in the bun, compound 50 was more effective in inhibiting the effect on TLR4. In other words, it was confirmed from the following experiment that the signaling pathway, oxidative stress, NF- ⁇ B activity, and inflammatory response were all suppressed (FIG. 2).
  • Cell viability assay was measured using EZ-cy Tox kit for the toxicity test and proper concentration of this compound.
  • Compounds 42 and 50 were measured at concentrations of 1 ⁇ M to 50 ⁇ M, respectively, and as a result, some cell deaths were observed at 50 ⁇ M (FIG. 3).
  • the concentration of 1,5,10 ⁇ M which is the most appropriate concentration, was selected as the experimental concentration.
  • ROS can be confirmed through a photograph, which means that the more fluorescence dye is expressed, the greater the amount of ROS, and it was confirmed that the fluorescence dye was significantly reduced in compounds 10 ⁇ M 42 and 5 ⁇ M and 10 ⁇ M 50 (Fig. 4b).
  • Western blotting boils the lysed sample in a cell with a loading buffer [0.125M Tris-Hcl, pH6.8, 4% SDS, 10% 2-mercaptoethanol and 0.2 bromophenol blue] for 1 to 5 minutes and loads onto the gel. . After SDS-PAGE using 10% acrylaminde by protein size, transfer to PVV membrane at 15V for 1 hour. After blocking for 1 hour with 5% skim milk, the primary antibody overnight and the secondary antibody are developed after 1-3 hours.
  • a loading buffer [0.125M Tris-Hcl, pH6.8, 4% SDS, 10% 2-mercaptoethanol and 0.2 bromophenol blue]
  • NF- ⁇ B a key factor involved in the regulation of inflammatory response and immune system
  • NF- ⁇ B consists of p50, p52, RelA (p65), RelB, c-Rel, and v-Rel. It has a variety of names.
  • the typical NF- ⁇ B activation pathway constitutes a signaling system essential for innate immunity.
  • the typical NF- ⁇ B activation pathway is a field in which many studies have been conducted until now, and the role of P65 and P50 is mainly played.
  • Phosphorylation of P65 has two main sites, Ser536 and Ser276. Looking at the following results it can be seen that the different aspects in Ser536 and Ser276 (Fig. 6).
  • the present inventors confirmed the translocation of NF- ⁇ B in the nucleus based on the above results, and performed an immunochemistry experiment using Alexa Fluor 488 Hoechst 33342.
  • the data were taken using confocal microscopy, Hoechst 33342 is blue as a dye for nuclei staining, and also stains portions of NF- ⁇ B in cells using NF- ⁇ B antibody and Alexa Fluor 488. Shown in green.
  • NF- ⁇ B is present only in the cytoplasm in the middle of the green part (nuclear part) in the untreated control group, and in the LPS group, the negative control group, the green part is generally cell-shaped. It can be seen that NF- ⁇ B translocation into the nucleus. Thus, NF- ⁇ B translocation into the nucleus due to LPS was found to decrease the concentration (translocation) of NF- ⁇ B by compounds 42 and 50 (Fig. 7).
  • Luciferase assay was performed once again to verify the effects of compounds 42 and 50 inhibiting NF- ⁇ B translocation into the nucleus. After transfection into cells using a luciferase reporter vector conjugated with an NF- ⁇ B binding site, LPS was treated to activate NF- ⁇ B. Luciferase is expressed by activated NF- ⁇ B, which luciferase reacts with luciferin to display phosphorescence, which is an indicator of how activated NF- ⁇ B is activated. As a result, it was confirmed that the activation of NF- ⁇ B increased by LPS was concentration-dependently inhibited by compounds 42 and 50 (FIG. 8).
  • NF- ⁇ B activation pathway a pathway that forms an essential signaling system for innate immunity, has been studied in many fields.
  • NF- ⁇ B dimers are inactivated in the cytoplasm by binding to IkBa that inhibits NF- ⁇ B activation.
  • IKB is degraded by the activity of IKK, and NF- ⁇ B in the cytoplasm moves into the nucleus and is activated. This process modulates the activity of IkB, which inhibits NF- ⁇ B in the cytoplasm, using the IKK complex.
  • IKK is composed of IKK ⁇ , IKK, and IKK ⁇ .
  • IKK-IKB-NF-KB Higher signaling pathways that regulate IKK in the NF- ⁇ B activation pathway
  • IKK-IKB-NF-KB include several higher signaling pathways such as ERK, p38, MAPK, and AKT.
  • Inhibition of ser536 of NF- ⁇ B in a concentration-dependent manner we confirmed the inhibitory effect of compounds 42 and 50 on the signaling pathway of AKT / PI3K.
  • compounds 42 and 50 inhibited AKT activity in a concentration-dependent manner, and that P-PTEN and NOX4, which regulates it, were also compound-dependent. And it was confirmed that it is suppressed by 50 (Fig. 10).
  • NOx4 is inhibited by compounds 42 and 50, thereby directly and indirectly inhibiting oxidative stress generated. Accordingly, it was confirmed in the present invention that the expression of inflammatory cytokines is reduced by blocking both PTEN activation and AKT / NF- ⁇ B signaling pathways.
  • the present inventors evaluated the efficacy against the same in vivo based on the experiment in vitro.
  • As an animal model C57BL / 6 mice were used, and one hour after feeding the compounds 42 and 50 to confirm that NF- ⁇ B was inhibited in the nucleus, 5 mg / kg LPS was IP dissected 1 hour later. It was.

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Abstract

La présente invention concerne de nouveaux composés présentant des effets antioxydants et anti-inflammatoires d'inhibition de l'activité des macrophages en raison de la concurrence avec LPS pour se lier à TLR4. L'invention porte également sur leur utilisation médicale. Les composés de l'invention présentent des activités antioxydantes et anti-inflammatoires, et peuvent donc être utilisés pour la prévention d'une maladie gériatrique ou le traitement de l'inflammation. En outre, les composés concurrencent LPS pour se lier à TLR4-MD2, en particulier, étant ainsi utilisés en tant que composition pharmaceutique utile dans la prévention et le traitement de la septicémie, des maladies métaboliques ou des maladies cardio-vasculaires, ou comme aliment de santé, en raison de la concurrence avec LPS pour se lier au récepteur TLR4-MD2.
PCT/KR2015/007655 2014-08-25 2015-07-23 Nouveaux composés présentant des activités antioxydantes et anti-inflammatoires en raison de la concurrence avec lps pour se lier à tlr4, et leur utilisation médicale WO2016032127A2 (fr)

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KR10-2014-0110853 2014-08-25
KR20140110853 2014-08-25
KR1020150101640A KR101686872B1 (ko) 2014-08-25 2015-07-17 Tlr4에 lps와의 경쟁적 결합을 통한 항산화 및 항염증 활성을 갖는 신규 화합물 및 이의 의학적 용도
KR10-2015-0101640 2015-07-17

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* Cited by examiner, † Cited by third party
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CN108276354A (zh) * 2018-02-06 2018-07-13 山西医科大学 Tlr4/md2抑制剂及其在抗炎药物中的应用
CN109563053A (zh) * 2016-06-28 2019-04-02 肯塔基大学研究基金会 前列腺素e合成酶抑制剂及使用其的方法
RU2786805C1 (ru) * 2022-07-06 2022-12-26 Федеральное государственное бюджетное образовательное учреждение высшего образования "Пермская государственная фармацевтическая академия" Министерства здравоохранения Российской Федерации Применение метил 5-бензоил-6-(4-фторфенил)-4-метокси-2-тиоксогексагидропиримидин-4-карбоксилата в качестве средства, обладающего противовоспалительной активностью

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6413936B1 (en) * 1995-10-30 2002-07-02 Glycorex Ab Glycomimetics as selectin antagonists and pharmaceuticals having antiinflammatory activity
WO2003074497A1 (fr) * 2002-03-01 2003-09-12 Pintex Pharmaceutical, Inc. Composes de modulation de pin1 et methodes d'utilisation associees
US20060025405A1 (en) * 2004-07-02 2006-02-02 Corcept Therapeutics, Inc. Modified pyrimidine glucocorticoid receptor modulators
WO2010136804A1 (fr) * 2009-05-28 2010-12-02 Aston University Dérivés de pyrimidine à utiliser en tant qu'antibiotiques

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6413936B1 (en) * 1995-10-30 2002-07-02 Glycorex Ab Glycomimetics as selectin antagonists and pharmaceuticals having antiinflammatory activity
WO2003074497A1 (fr) * 2002-03-01 2003-09-12 Pintex Pharmaceutical, Inc. Composes de modulation de pin1 et methodes d'utilisation associees
US20060025405A1 (en) * 2004-07-02 2006-02-02 Corcept Therapeutics, Inc. Modified pyrimidine glucocorticoid receptor modulators
WO2010136804A1 (fr) * 2009-05-28 2010-12-02 Aston University Dérivés de pyrimidine à utiliser en tant qu'antibiotiques

Non-Patent Citations (20)

* Cited by examiner, † Cited by third party
Title
ADZHIBEKYAN, A. C. ET AL.: "Some 5- and N-arylsubstituted Barbiturates and their Biological Properties", ARMYANSKII KHIMICHESKII ZHURNAL, vol. 28, no. 9, 1975, pages 741 - 746, XP009055735 *
ARPYAN, A. A. ET AL.: "Synthesis of Some Substituted 5-(P-Alkoxybenzyl) -Pyrimidines", ARMYANSKII KHIMICHESKII ZHURNAL, vol. 20, no. 1, 1967, pages 61 - 67 *
DATABASE STN 10 February 2009 (2009-02-10), accession no. N 1103939-46-3 *
DATABASE STN 13 April 2008 (2008-04-13), accession no. N 1013999-95-5 *
DATABASE STN 17 July 2006 (2006-07-17), accession no. N 893626-83-0 *
DATABASE STN 2 February 2001 (2001-02-02), accession no. N 319489-78-6 *
DATABASE STN 21 October 2009 (2009-10-21), accession no. N 1189292-50-9 *
DATABASE STN 27 October 2009 (2009-10-27), accession no. N 1190251-16-1 *
DATABASE STN 7 January 2002 (2002-01-07), accession no. N 380544-79-6 *
DATABASE STN 8 August 2008 (2008-08-08), accession no. N 1039676-15-7 *
DATABASE STN 9 September 2008 (2008-09-09), accession no. N 1047976-45-3 *
DATABASE STN EXPRESS 29 January 2002 (2002-01-29), accession no. N 387830-20-8 *
GOUDGAON, N. M ET AL.: "Synthesis and Antimicrobial Activities of Novel 5-Substituted Pyrimidin-2, 4, 6-Triones", J OURNAL OF THE INDIAN CHEMICAL SOCIETY, vol. 87, no. 6, 2010, pages 743 - 748 *
GRIGORYAN, L. A. ET AL.: "Arylsulphonic Acid Derivatives. IV. Sulfamido Derivatives of 5-(P-Alkoxybenzyl)Pyrimidines", ARMYANSKII KHIMICHESKII ZHUMAL, vol. 28, no. 9, 1975, pages 704 - 708 *
JURSIC, B. S. ET AL.: "Mono C-alkylation and Mono C-benzylation of Barbituric Acids through Zinc/Acid Reduction of Acyl, Benzylidene, and Alkylidene Barbiturate Intermediates", TETRAHEDRON LETTERS, vol. 44, no. 10, 2003, pages 2203 - 2210, XP004410042, DOI: doi:10.1016/S0040-4039(03)00111-4 *
KOZLOV, N. ET AL.: "Synthesis of Arylmethylpyrimidinetriones and Pyrimidoquinolinediones with Fluorescent and Nonlinear-Optical Properties", RUSSIAN JOURNAL OF APPLIED CHEMISTRY, vol. 80, no. 7, 2007, pages 1101 - 1104 *
KUJUNDZIC, N. ET AL.: "Synthesis and Antibacterial Effect of Derivatives of 5-(3,4,5-Trimethoxybenzyl)Pyrimidine, Tetrahydropyrimidine, -Hexahydropyrimidine and -Hydantoin", CROATICA CHEMICA ACTA, vol. 61, no. 1, 1988, pages 121 - 135, XP009070565 *
MCADOO, M. H. ET AL.: "Inhibition of the Cathepsin D-Type Proteinase of Macrophages by Pepstatin, a Specific Pepsin Inhibitor, and Other Substances", INFECTION AND IMMUNITY, vol. 7, no. 4, 1973, pages 655 - 665 *
SAWANT, R. L. ET AL.: "QSAR and Docking Study for Rate of Mushroom Tyrosinase Inhibition by Some 5-Benzylidene Barbiturate Derivatives", INTERNATIONAL JOURNAL OF DRUG DESIGN AND DISCOVERY, vol. 4, no. 2, 2013, pages 1077 - 1082 *
VIEIRA, A. A. ET AL.: "Synthesis and In Vivo Evaluation of 5-(Chloro-5-Benzyl-Barbiturates as New Central Nervous System Depressants", JOURNAL OF THE BRAZILIAN CHEMICAL SOCIETY, vol. 22, no. 2, 2011, pages 364 - 371 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109563053A (zh) * 2016-06-28 2019-04-02 肯塔基大学研究基金会 前列腺素e合成酶抑制剂及使用其的方法
CN108276354A (zh) * 2018-02-06 2018-07-13 山西医科大学 Tlr4/md2抑制剂及其在抗炎药物中的应用
CN108276354B (zh) * 2018-02-06 2020-02-18 山西医科大学 Tlr4/md2抑制剂及其在抗炎药物中的应用
RU2786805C1 (ru) * 2022-07-06 2022-12-26 Федеральное государственное бюджетное образовательное учреждение высшего образования "Пермская государственная фармацевтическая академия" Министерства здравоохранения Российской Федерации Применение метил 5-бензоил-6-(4-фторфенил)-4-метокси-2-тиоксогексагидропиримидин-4-карбоксилата в качестве средства, обладающего противовоспалительной активностью

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