CN114478511B - Benzoxazole compound and preparation method, pharmaceutical composition and application thereof - Google Patents
Benzoxazole compound and preparation method, pharmaceutical composition and application thereof Download PDFInfo
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Abstract
The invention discloses a benzoxazole compound, a preparation method, a pharmaceutical composition and application thereof. The compound has a structure shown in a formula (I), and comprises pharmaceutically acceptable salts thereof. The benzoxazole compound and the pharmaceutical composition thereof can effectively antagonize P2Y 6 The receptor, the in vitro antagonism rate is optimally up to 100%, IC 50 The values optimally reached picomolar concentration levels; shortening of colon length, weight loss, and elevation of DAI score in mice can be inhibited in vivo. Can be used for preparing and P2Y in therapy 6 The medicine for treating receptor-related inflammatory diseases can exert the medicine effect on the cellular level and animal level, has wide application, and the synthesis method of the compound is simple and easy to implement.
Description
Technical Field
The invention relates to a benzoxazole compound, a preparation method, a pharmaceutical composition and application thereof, in particular to a compound capable of being prepared into P2Y 6 Benzoxazole compounds of receptor antagonist medicaments, and a preparation method, a pharmaceutical composition and application thereof.
Background
P2Y 6 Receptor (P2Y) 6 R) is eight subtypes of the P2Y receptor family (P2Y) 1 R、P2Y 2 R、P2Y 4 R、 P2Y 6 R、P2Y 11 R、P2Y 12 R、P2Y 13 R and P2Y 14 R) expressed in immune organs, cardiovascular system, nervous tissue and other organs and tissues, P2Y 6 The endogenous ligand of the receptor is an extracellular nucleotide molecule through which the receptor is selectedSelective agonist UDP specifically activates phospholipase C (PLC) up-regulating intracellular Ca 2+ The concentration achieves the purposes of transmitting signals among cells and regulating various physiological functions of the cells. Current research shows that when P2Y 6 Upon receptor agonism, neutrophil and macrophage recruitment and chemotaxis are promoted and a variety of inflammatory cytokines, chemokines and mast cell mediators are released. By P2Y 6 Studies in a mouse model of receptor knockout, results confirm P2Y 6 The receptor is involved in the occurrence and development of cardiovascular diseases, respiratory tract inflammation, gastrointestinal tract inflammation and other diseases.
P2Y 6 The receptor antagonist has good innovation and application prospect in the field of medicine development of inflammatory diseases related to inflammatory bowel diseases, atherosclerosis, neurodegenerative diseases, rheumatism and the like. It is noted that P2Y is blocked alone in dextran sodium sulfate DSS induced colitis models 6 The receptor can protect the intestinal tract from inflammation. Extracellular nucleotide signals on the surface of intestinal epithelial cells play an important role in the development of intestinal inflammation and possibly in inflammatory bowel disease IBD. Regulation of P2Y 6 The signal may be a new potential target for the treatment of IBD. P2Y 6 Receptors are widely distributed in various tissues and immune cells and are involved in inflammatory reactions by regulating the expression and secretion of cytokines and pro-inflammatory molecules, and have become potential therapeutic targets for the treatment of a variety of cardiovascular diseases including atherosclerosis. Its specific regulatory mechanism is silencing P2Y 6 The receptor can inhibit the expression of PLC beta, thereby inhibiting intracellular Ca 2+ Up-regulation of NLRP3 inflammatory bodies caused by elevated concentrations and release of inflammatory factors such as IL-1 beta and IL-8, thereby inhibiting cell apoptosis. Cell apoptosis is programmed necrosis of cells mediated by gasdermin, accompanied by explosive release of inflammatory factors such as IL-1 beta. Macrophage pyrosis mediated by NLRP3 inflammatory body activation plays a decisive role in atherosclerosis. Microglial cells in central nerve cells play a key role in neurodegenerative diseases, and peripheral blood mononuclear cells P2Y in parkinsonism patients are found by research 6 Receptor levels are significantly elevated, while inhibition of P2Y in vitro experiments 6 Receptor or knock-out P2Y 6 The receptor can reduce the release of inflammatory factors such as microglial cells TNF-alpha, iNOS, IL-6, COX-2 and the like, and increase the survival rate of nerve cells cultured from the supernatant. Studies indicate that P2Y 6 Receptors can be used as an effective index of Parkinson's disease, and P2Y is blocked 6 The receptor may treat parkinson's disease by alleviating neuroinflammation caused by microglia. Gout is a crystal-related joint disease caused by deposition of sodium urate (MSU) on joints, and belongs to metabolic rheumatism. The surface of neutrophils is rich in P2Y 6 Receptors, and neutrophils, when relieved of sodium urate (MSU) crystals in gout patients, produce a severe inflammatory response in the joint. There are studies that have found a large number of Neutrophil Extracellular Traps (NETs) in synovial fluid of gout patients. Blocking P2Y 6 The receptor can inhibit neutrophil activation, reduce IL-8 release, block sodium urate-induced NETs formation, relieve gout symptom, and prompt P2Y 6 The receptor antagonist can be used as a novel gout therapeutic drug.
Existing P2Y 6 The receptor antagonists have few structural types, and have the defects of low bioavailability, poor stability and the like, and the P2Y with the best reported activity is known 6 The receptor antagonist MRS2578 can only be used as a tool medicine, so that the high-activity P2Y with a brand new structure is developed 6 Receptor antagonists are of great importance.
Disclosure of Invention
The invention aims to: aiming at the defects of low bioavailability, poor stability and the like of the existing compounds, the invention aims to provide a benzoxazole compound capable of effectively antagonizing a P2Y6 receptor, a preparation method, a pharmaceutical composition and application thereof.
The technical scheme is as follows: as a first aspect to which the present invention relates, the benzoxazole compound of the present invention has a structure of formula (I), said benzoxazole compound comprising a pharmaceutically acceptable salt thereof:
wherein:
R 1 selected from benzene ring, substituted benzene ring or 5-6 membered heterocycle;
R 2 selected from substituted or unsubstituted C 1 ～C 6 Alkyl, C 1 ～C 6 Alkoxy, hydrogen, 5-6 membered heterocycle, C 1 ～C 6 Alkenyl, cyano, hydroxy or halogen.
Benzoxazoles are an important pharmacophore in modern drug discovery, and a large number of benzoxazoles have been successfully developed. Researches show that the benzoxazole heterocyclic compound has good biological activities of resisting inflammation and bacteria, treating atherosclerosis, resisting viruses and the like, has wide biological activities, and has low toxicity, high bioavailability, good biocompatibility and curative effect. By utilizing the principle of active superposition, the activity of the benzoxazole group is greatly improved after the benzoxazole group is introduced into a plurality of small molecular medicines.
Preferably, in the above structure:
R 1 selected from pyrrole ring, pyrazole ring, imidazole ring, thiophene ring, thiazole ring, oxazole ring, furan ring, pyran ring, piperidine ring, piperazine ring, pyridine ring, pyrazine ring, benzene ring or substituted benzene ring;
R 2 selected from hydrogen, methyl, ethyl, isopropyl, tert-butyl, methoxy, methoxyethyl, allyl, cyano, phenolic hydroxyl, fluorine atom, chlorine atom, nitro or trifluoromethyl.
Further preferably, in the above structure:
R 1 selected from pyrrole ring, furan ring, benzene ring or substituted benzene ring;
R 2 selected from hydrogen, methyl or chlorine atoms.
Still more preferably, in the above structure:
R 1 selected from pyrrole ring, furan ring, benzene ring or 4-methyl substituted phenyl;
R 2 selected from hydrogen, 5-methyl substituents, 6-methyl substituents, 7-methyl substituents or 6-chloro substituents.
Most preferably, the benzoxazole compound is selected from any one of the following compounds:
the pharmaceutically acceptable salt of the above benzoxazole compound is a salt formed by the above-mentioned oxazole compound and an acid, wherein the acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.
As a second aspect of the present invention, the method for producing the benzoxazole compound comprises:
the compound (II) and the compound (III) are subjected to cyclization reaction to obtain a compound (I);
wherein R is 1 、R 2 Is as defined above;
the specific synthesis method is as follows:
and (3) salifying the corresponding acid with the compound (I) prepared by the method to obtain the pharmaceutically acceptable salt of the benzoxazole compound.
As a third aspect of the present invention, the pharmaceutical composition of the present invention comprises the above-mentioned benzoxazole compound and a pharmaceutically acceptable carrier.
The benzoxazole compound can be added with pharmaceutically acceptable carriers to prepare common medicinal preparations such as tablets, capsules, syrup, suspending agents or injection, and the preparations can be added with common medicinal auxiliary materials such as perfume, sweetener, liquid/solid filler, diluent and the like.
As a fourth aspect of the present invention, the benzoxazole compound and the pharmaceutical composition thereof can be prepared as P2Y 6 Receptor antagonist drugs for treatment and P2Y 6 The receptor-related inflammatory diseases can be used for treating inflammatory bowel disease, atherosclerosis, neurodegenerative diseases, rheumatism, etc.
The beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages:
(1) The benzoxazole compound and the pharmaceutical composition thereof can effectively antagonize P2Y 6 The receptor, the in vitro antagonism rate is optimally up to 100%, IC 50 The values optimally reached picomolar concentration levels; can inhibit the reduction of colon length, weight reduction and DAI score increase of mice in vivo;
(2) The benzoxazole compound and the pharmaceutical composition thereof have wide application and can be prepared and treated as P2Y 6 A medicament for the treatment of a receptor-related inflammatory disease; the medicine can exert the medicine effect at the cellular level and the animal level, has more excellent treatment effect, and can optimally reach the picomolar concentration level;
(3) The preparation method of the compound is simple and easy to implement.
Drawings
FIG. 1 is a graph showing the effect of compound I-1 on the body weight of mice with DSS-induced colitis;
FIG. 2 is a graph showing the effect of compound I-1 on DAI index in mice with DSS-induced colitis;
FIG. 3 is a graph showing the effect of compound I-1 on DSS-induced colonic length in a colitis mouse;
FIG. 4 shows the results of colon histopathological staining.
Detailed Description
The technical scheme of the invention is further described below by referring to examples.
Example 1: synthesis of 2- (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) benzo [ d ] oxazole
2.8g of potassium tert-butoxide are placed in 11mL of THF to form a suspension, which is cooled in an ice bath at 0 ℃; 1.1g of acetylfuran and 2.7mL of diethyl oxalate were dissolved in 11mL of ethylene glycol dimethyl ether, and this solution was added dropwise to a suspension of potassium t-butoxide at 0 ℃; the mixture was stirred at room temperature for 1 hour. After all the acetylfuran was consumed as monitored by TLC, 7.5mL of 1M HCl solution was added. The crude product was extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, dried under reduced pressure, purified by silica gel column chromatography, petroleum ether: ethyl acetate (4:1) to give ethyl 4- (furan-2-yl) -2, 4-dioxobutyrate.
1.4g of ethyl 4- (furan-2-yl) -2, 4-dioxobutyrate was dissolved in ethanol, 0.9g of tert-butylhydrazine hydrochloride was added, and the mixture was stirred at room temperature overnight. When the reaction was complete, the crude product was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and dried under reduced pressure. The product was purified by column chromatography on silica gel using petroleum ether: ethyl acetate (50:1-20:1) to give ethyl 1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazole-3-carboxylate.
200mg of ethyl 1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazole-3-carboxylate were dissolved in 3mL of tetrahydrofuran, which was then added dropwise to a slurry of lithium aluminum hydride (50 mg) in 10mL of tetrahydrofuran at 0 ℃. After 30 minutes at this temperature, the reaction was heated to reflux for 2 hours. The reaction was cooled to room temperature, and 10mL of ethyl acetate was added to the reaction. To the reaction solution was added 5N sodium hydroxide until white precipitation occurred. The mixture was filtered, and the filtrate was extracted with ethyl acetate and water. The organic layer was dried over magnesium sulfate, filtered and dried under reduced pressure to give crude (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) methanol.
188mg of (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) methanol was dissolved in DCM, 434mg of Dess-Martin reagent was added under ice bath, stirred at room temperature for 1H, quenched with saturated sodium carbonate, filtered through celite, the filtrate extracted with ethyl acetate, dried over anhydrous sodium sulfate, and spun dry under reduced pressure to give crude 1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazole-3-carbaldehyde.
80mg of 1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazole-3-carbaldehyde, 60mg of o-aminophenol, 80mg sodium bisulphite were dissolved in ethanol: water = 2:1, and refluxing for 1h. Ethyl acetate extraction, drying over anhydrous sodium sulfate, spin-drying under reduced pressure, purification by thin layer chromatography, petroleum ether: ethyl acetate (3:1) to give the pure target product.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d 6 )δ7.11(d,J＝1.7Hz,1H),6.98(td,J＝8.2,2.1 Hz,2H),6.62–6.58(m,2H),6.36(s,1H),5.99(d,J＝3.3Hz,1H),5.87(dd,J＝3.3,1.9Hz,1H),0.73(s,9H).
13 C NMR(101MHz,DMSO-d 6 )δ158.13,150.29,144.52,142.91,141.68, 137.22,134.27,125.91,125.31,120.18,113.36,112.27,112.23,111.46,62.82,40.64,40.43,40.22,40.01,39.80,39.59,39.39,29.97.
example 2: synthesis of 2- (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) -5-methylbenzo [ d ] oxazole
The synthesis is described in example 1 starting from o-amino-p-cresol.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.92(s,1H),7.66(d,J＝8.3Hz,1H),7.57(s, 1H),7.23(d,J＝8.3Hz,1H),7.15(s,1H),6.80(d,J＝3.3Hz,1H),6.68(d,J＝2.6Hz,1H),2.44(s,3H),1.53(s,9H).
13 C NMR(101MHz,DMSO-d6)δ158.21,148.51,144.48,142.95,141.90, 137.33,134.68,134.21,126.86,119.96,113.31,112.25,112.12,110.82,62.76,40.62,40.42,40.21,40.00,39.79,39.58,39.37,34.94,31.60,30.28,29.96,21.46.
example 3: synthesis of 2- (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) -5-chlorobenzo [ d ] oxazole
The synthesis is described in example 1 starting from 4-chloro-2-aminophenol.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ8.07(dd,J＝7.5,1.6Hz,1H),7.84–7.72(m, 2H),7.40(dd,J＝7.5,1.5Hz,1H),7.27–7.16(m,2H),6.98(t,J＝7.5Hz,1H),1.46 (s,10H).
13 C NMR(101MHz,DMSO-d6)δ158.07,152.16,150.75,142.66,140.52, 131.95,131.20,130.80,126.55,119.50,112.14,111.51,104.50,60.49,28.94.
example 4: synthesis of 2- (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) -4-methylbenzo [ d ] oxazole
Starting from 2-amino-3-methylphenol, the synthesis is described in example 1.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.91(d,J＝1.7Hz,1H),7.57(d,J＝8.0Hz, 1H),7.29(t,J＝7.8Hz,1H),7.19(d,J＝7.5Hz,1H),7.16(s,1H),6.79(d,J＝3.3Hz,1H),6.67(dd,J＝3.3,1.9Hz,1H),2.55(s,3H),1.53(s,9H).
13 C NMR(101MHz,DMSO-d6)δ157.45,150.00,144.49,142.98,140.90, 137.34,134.24,130.27,125.67,125.61,113.29,112.29,112.08,108.69,62.77,40.63, 40.42,40.22,40.01,39.80,39.59,39.38,29.98,16.76.
example 5: synthesis of 2- (1- (tert-butyl) -5- (furan-2-yl) -1H-pyrazol-3-yl) -6-methylbenzo [ d ] oxazole
Starting from 6-amino-m-cresol, the synthesis is described in example 1.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.91(s,1H),7.63(d,J＝8.1Hz,1H),7.60(s, 1H),7.21(d,J＝8.1Hz,1H),7.13(s,1H),6.79(d,J＝3.4Hz,1H),6.67(d,J＝2.6Hz,1H),2.45(s,3H),1.52(s,9H).
13 C NMR(101MHz,DMSO-d6)δ157.63,150.57,144.50,142.96,139.50, 137.36,135.93,134.21,126.42,119.57,113.32,112.26,112.05,111.42,62.75,40.64,40.43,40.22,40.01,39.80,39.60,39.39,31.62,29.97,21.78.
example 6: synthesis of 2- (1- (tert-butyl) -5- (1H-pyrrol-2-yl) -1H-pyrazol-3-yl) benzo [ d ] oxazole
Starting from 2-acetylpyrrole, the synthesis is described in example 1.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ11.26(s,1H),7.82–7.75(m,2H),7.43– 7.38(m,2H),6.97(s,1H),6.91(q,J＝2.4Hz,1H),6.26(dt,J＝3.8,1.8Hz,1H), 6.16(q,J＝2.8Hz,1H),1.48(s,9H).
13 C NMR(101MHz,DMSO-d6)δ158.57,150.28,141.76,137.79,136.86, 125.77,125.24,120.10,119.89,119.43,112.08,111.77,111.43,108.58,62.61,30.40.
example 7: synthesis of 2- (1- (tert-butyl) -5- (1H-pyrrol-2-yl) -1H-pyrazol-3-yl) -5-methylbenzo [ d ] oxazole
Starting from 2-acetylpyrrole and o-amino-p-cresol, the synthesis is described in example 1.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ11.28–11.24(m,1H),7.66(d,J＝8.3Hz, 1H),7.57(d,J＝1.6Hz,1H),7.22(dd,J＝8.4,1.6Hz,1H),6.95(s,1H),6.92(q,J＝2.6Hz,1H),6.26(dt,J＝3.9,1.7Hz,1H),6.16(q,J＝2.7Hz,1H),2.44(s,3H),1.47 (s,9H).
13 C NMR(101MHz,DMSO-d6)δ158.65,148.51,141.99,137.73,136.98, 134.61,126.71,119.91,119.41,111.98,111.74,110.80,108.57,62.56,31.61,30.39,21.47.
example 8: synthesis of 2- (1- (tert-butyl) -5- (1H-pyrrol-2-yl) -1H-pyrazol-3-yl) -5-chlorobenzo [ d ] oxazole
Starting from 2-acetylpyrrole and 4-chloro-2-aminophenol, the synthesis is described in example 1.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.84–7.72(m,2H),7.40(dd,J＝7.5,1.5Hz, 1H),7.14(s,1H),7.00(dd,J＝7.5,1.5Hz,1H),6.49(dd,J＝7.5,1.6Hz,1H),6.33(t, J＝7.5Hz,1H),1.43(s,10H).
13 C NMR(101MHz,DMSO-d6)δ158.07,150.75,140.52,133.71,131.95, 130.80,126.55,125.82,124.54,119.50,112.14,111.43,107.23,107.06,60.49,28.94.
example 9: synthesis of 2- (1- (tert-butyl) -5-phenyl-1H-pyrazol-3-yl) -5-chloro-1H-benzo [ d ] imidazole
The synthesis is described in example 1 using acetophenone as starting material.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.82–7.76(m,2H),7.49(s,5H),7.43–7.38 (m,2H),6.88(s,1H),1.47(s,9H).
13 C NMR(101MHz,DMSO-d6)δ158.56,150.31,145.02,141.77,137.10, 132.87,130.89,129.60,128.60,125.77,125.22,120.12,111.42,110.49,63.05,31.11.
example 10: synthesis of 2- (1- (tert-butyl) -5-phenyl-1H-pyrazol-3-yl) -5-chloro-1H-4-methyl-benzo [ d ] imidazole
The synthesis is described in example 1 starting from acetophenone and 6-amino-m-cresol.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.66–7.61(m,2H),7.49(s,5H),7.22(dd,J ＝8.2,1.5Hz,1H),6.85(s,1H),2.47(s,3H),1.48(s,9H).
13 C NMR(101MHz,DMSO-d6)δ158.05,150.59,144.96,139.58,137.23, 135.78,132.91,130.89,129.59,128.59,126.33,119.51,111.39,110.33,62.98,31.12,21.77.
example 11: synthesis of 2- (1- (tert-butyl) -5-4-methylphenyl-1H-pyrazol-3-yl) -5-chloro-1H-4-methyl-benzo [ d ] imidazole
The synthesis is described in example 1 starting from p-methylacetophenone and 6-amino-m-cresol.
The nuclear magnetic data are as follows:
1 H NMR(400MHz,DMSO-d6)δ7.59–7.43(m,2H),7.24–7.16(m,1H),2.43 (s,1H),2.33(t,J＝1.2Hz,1H),1.35(s,3H).
13 C NMR(101MHz,DMSO-d6)δ157.74,150.23,149.59,137.74,137.12, 135.07,131.02,130.89,129.03,127.93,127.15,118.86,109.90,104.44,60.32,28.94,21.42,21.21.
example 12: compound pair P2Y 6 Receptor in vitro antagonistic activity assay
Human P2Y to be constructed earlier 6 R stable HEK293 cells were cultured in DMEM medium (containing 10% fetal calf serum, 100U/mL penicillin and 100. Mu.g/mL streptomycin) and inoculated into 6-well plates at a density of 5X 10 prior to the experiment 5 cells/ml, cells at 37℃and 95% O 2 、5％CO 2 Culturing under humidity. Serum-free medium is changed before the experiment to starve for 12 hours, 1 mu M compound is added into each hole, after 30 minutes of reaction, 10 mu M UDP is added for incubation for 12 hours, and then a sample is collectedDetecting the content of inositol 3 phosphate (IP 3).
The inositol 3 phosphate (IP 3) enzyme-linked immunosorbent assay kit adopts a competition ELISA method. The IP3 antigen is coated on an ELISA plate, and during experiments, IP3 in a sample or a standard substance competes with the coated IP3 for binding sites on the biotin-labeled anti-IP 3 monoclonal antibody, and free components are washed off. Horseradish peroxidase-labeled avidin is added, biotin is specifically combined with avidin to form an immune complex, and free components are washed away. Chromogenic substrate (TMB) was added and the TMB appeared blue under the catalysis of horseradish peroxidase, and turned yellow after addition of stop solution. And (3) measuring an OD value at a wavelength of 450nm by using an enzyme-labeled instrument, wherein the concentration of IP3 is inversely proportional to the OD450 value, and calculating the concentration of IP3 in the sample by drawing a standard curve.
And finally, calculating the average OD value of each group of compound holes. Drawing a standard curve of a four-parameter logic function on double-logarithmic-coordinate paper by taking the concentration as an abscissa and the OD value as an ordinate; the concentration of IP3 in the sample was calculated from the standard curve. Experiments were repeated three times, averaged and the compound pair P2Y calculated 6 IC of R 50 The experimental results are shown in table 1.
Table 1 pair P2Y 6 Results of in vitro antagonistic Activity test
| Compounds of formula (I) | ％inhibition at 10μM | hP2Y 6 R IC 50 (nM) |
| I-1 | 118.06 | 0.64 |
| I-2 | 72.28 | 102.2 |
| I-3 | 82.64 | 65.09 |
| I-4 | 100.37 | 37.23 |
| I-5 | 104.66 | 43.19 |
| I-6 | 108.63 | 42.51 |
| I-7 | 68.45 | 96.45 |
| I-8 | 86.39 | 73.38 |
| I-9 | 98.56 | 95.18 |
| I-10 | 102.17 | 68.77 |
| I-11 | 86.34 | 44.87 |
As can be seen from Table 1, all test compounds pair P2Y 6 R has inhibition effect, the inhibition rate is more than 65%, and the highest inhibition rate reaches 100%; IC (integrated circuit) 50 The values are all at nanomolar levels, with compound I-1 reaching even picomolar levels.
Example 13: in vivo efficacy test of compounds on inflammatory bowel disease
Male C57BL/6 mice (22.+ -.2 g) of 6-8 weeks of age were randomly divided into 5 groups of 6 animals each. The method comprises the following steps of: control, DSS, compound I-1 (50. Mu.M) and 5-aminosalicylic acid (5-ASA) (30 mg/kg) were administered by rectal administration in a volume of 100. Mu.L per day. Model and dosing groups mice were free to drink 3% DSS solution for 7 days to induce colitis, with freshly prepared DSS solution changed every 2 days. The Control group had free access to distilled water for 7 days and mice were sacrificed for material sampling on day 8. The mice of each group were weighed daily during the administration period (fig. 1), and observed for hair color, mental state, stool consistency, and rectal bleeding. The model group and each dosing group were scored for colitis disease activity using disease activity index (Disease activity index, DAI) (fig. 2).
DAI scoring index: the sum of the percentage of weight loss (weight not changing to 0 score, 1-5% 1 score, 5-10% 2 score, 10-15% 3 score, > 15% 4 score), stool hardness (normally 0 score, slightly moist but not adhering to the perianal area, diarrhea 4 score) and rectal bleeding (normally 0 score, occult blood positive 2 score, macroscopic bloody stool 4 score) is DAI. The DAI composite score is between 0 and 4 points, with a score of 0 representing normal and a score of 4 representing maximum activity of inflammation.
After the modeling was completed, the mice were sacrificed by cervical removal, the entire colon from the anus to the distal cecum was removed, the length of the colon was measured (fig. 3), and then the colon was rinsed in pre-chilled saline and the water was blotted on filter paper, and a portion of the distal colon was used for pathology detection (fig. 4).
Experimental results show that I-1 has good improvement results on the phenotype of the acute ulcerative colitis disease of mice induced by DSS, such as inhibition of DSS-induced shortening of colon length, weight reduction and DAI score increase of the mice. In addition, I-1 has relieving effect on damage of colon tissue intestinal wall and loss of goblet cells induced by DSS, and the results show that I-1 has good curative effect on inflammatory bowel disease.
As can be seen from the above examples, the benzoxazole derivative prepared according to the invention has better therapeutic effect on P2Y 6 Receptor-related inflammatory diseases such as inflammatory bowel disease, atherosclerosis, neurodegenerative diseases, rheumatism, etc.
Claims (10)
1. A benzoxazole compound having the structure of formula (I), said compound comprising a pharmaceutically acceptable salt thereof:
wherein:
R 1 selected from pyrrole ring, furan ring, benzene ring or 4-methyl substituted phenyl;
R 2 selected from C 1 ～C 6 Alkyl, hydrogen or halogen.
2. The benzoxazole compound according to claim 1, wherein in said structure:
R 2 selected from hydrogen, methyl or chlorine atoms.
3. The benzoxazole compound according to claim 1 or 2, characterized in that in said structure:
R 2 selected from hydrogen, 5-methyl substituents, 6-methyl substituents, 7-methyl substituents or 6-chloro substituents.
4. The benzoxazole compound according to claim 1 or 2, characterized in that it is selected from any one of the following compounds:
5. the benzoxazole compound according to claim 1 or 2, characterized in that said pharmaceutically acceptable salt is a salt of said benzoxazole compound with an acid, said acid being hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.
6. A process for producing a benzoxazole compound according to any one of claims 1 to 5, characterized in that said process comprises:
wherein R is 1 、R 2 Is as defined in any one of claims 1 to 4;
and (3) salifying the corresponding acid with the compound (I) prepared by the method to obtain the pharmaceutically acceptable salt of the benzoxazole compound.
7. A pharmaceutical composition comprising a benzoxazole compound according to any one of claims 1-5 and a pharmaceutically acceptable carrier.
8. A process for preparing P2Y comprising preparing a benzoxazole compound according to any one of claims 1 to 5 or a pharmaceutical composition according to claim 7 6 Application of receptor antagonist in medicineIs used.
9. The use according to claim 8, wherein the medicament is for the treatment of P2Y 6 Receptor-related inflammatory diseases.
10. The use according to claim 9, wherein said and P2Y 6 The receptor-related inflammatory disease is selected from inflammatory bowel disease, atherosclerosis, neurodegenerative diseases, rheumatism.
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