CN113816934B - Synthesis method of disubstituted chromanone and application of disubstituted chromanone in treatment of pulmonary inflammation such as COPD (chronic infectious disease) - Google Patents

Synthesis method of disubstituted chromanone and application of disubstituted chromanone in treatment of pulmonary inflammation such as COPD (chronic infectious disease) Download PDF

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CN113816934B
CN113816934B CN202111129637.7A CN202111129637A CN113816934B CN 113816934 B CN113816934 B CN 113816934B CN 202111129637 A CN202111129637 A CN 202111129637A CN 113816934 B CN113816934 B CN 113816934B
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disubstituted
chroman
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CN113816934A (en
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姚宏亮
李刚
潘文俊
关文
张亚莉
王华敏
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Institute of Zoology of Guangdong Academy of Sciences
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Abstract

The invention discloses a synthesis method of disubstituted chromanone and application thereof in treating pulmonary inflammation such as COPD and the like. 4', 7-disubstituted chroman-4-ones having the chemical structure shown in formula (a), wherein R 1 The radicals are isobutyl, n-butyl, cyclopentylmethyl, cyclopropylmethyl, cyclohexylmethyl, benzyl, 4-methoxybenzyl, tert-butyldimethylsilyl, (tetrahydro-2H-pyran-4-yl) methyl, 2- (piperidin-1-yl) ethyl, 2-morpholinoethyl, 2- (methyl (phenyl) amino) ethyl or 2- (1H-imidazol-1-yl) ethyl, R 2 The group is tert-butyldimethylsilyl, isobutyl or cyclopentylmethyl. The invention synthesizes 4', 7-disubstituted chroman-4-one, and experiments show that the 4', 7-disubstituted chroman-4-one can effectively inhibit inflammatory reaction of lung. Therefore, can be used for inhibiting lung inflammatory reaction and has wide application.

Description

Synthesis method of disubstituted chromanone and application of disubstituted chromanone in treatment of pulmonary inflammation such as COPD (chronic infectious disease)
Technical field:
the invention belongs to the field of biological medicine, and in particular relates to 4', 7-disubstituted chroman-4-one, a synthesis method thereof and application thereof in preparing medicines for treating pulmonary inflammation such as COPD (chronic infectious disease).
The background technology is as follows:
pneumonia refers to inflammation in the distal lung, i.e., the lung interstitium, alveolar space, and terminal airways. Factors causing inflammation are mainly pathogenic microorganisms such as bacteria, parasites, fungi, viruses, etc., or chemoallergies, radiation, etc. The clinical symptoms of pneumonia are mainly cough, fever, bloody sputum or expectoration in the sputum, and often accompanied by dyspnea or chest distress and the like. Pneumonia can be classified into viral pneumonia, fungal pneumonia, bacterial pneumonia, mycoplasma pneumonia, physicochemical pneumonia, allergic pneumonia, other pathogen pneumonia, immune pneumonia and the like according to etiology, and the most common pneumonia is bacterial pneumonia, accounting for more than 70% of adult pneumonia. When pathogens invade lung tissue, the signaling pathways of Mitogen Activated Protein Kinase (MAPK), nuclear transcription factor- κB, etc. of the body are activated, triggering inflammatory reactions for immunization. In inflammatory response, immune cells such as alveolar macrophages, neutrophils and lymphocytes are activated by the signal transduction pathway, and inflammatory mediators such as tumor necrosis factor, interleukin, interferon, growth factor and chemotactic factor are released, for example, TNF-alpha, IL-6, IFN-gamma, IL-1 beta, IL-12, IL-18 and the like, which perform feedback regulation on the inflammatory response and the signal transduction pathway. Pulmonary inflammation is an important component of various acute and chronic respiratory diseases such as COPD, asthma, ARDS and the like, and can cause patients to suffer from mucous hypersecretion, airway obstruction, emphysema, pulmonary edema and other diseases, and complications of other organs and tissues, and even systemic diseases, which seriously affect life.
Flavonoid compounds are widely used in the treatment of pulmonary inflammation due to their unique structural features. Corresponding to the characteristics of pathological processes of occurrence and development of inflammatory reactions of lung tissues, the mechanism of flavonoid compounds for resisting pneumonia mainly comprises the following ways: the preparation method can regulate MAPK, NF- κB and other signal transduction pathways to influence the accumulation of inflammatory cells in lung, inhibit the release of cytokines and reduce the generation of pro-inflammatory mediators so as to inhibit the occurrence and development of inflammatory reactions.
The invention comprises the following steps:
the invention aims to provide 4', 7-disubstituted chroman-4-one, a synthesis method thereof and application thereof in preparing medicines for treating pulmonary inflammation such as COPD and the like.
The chemical structure of the 4', 7-disubstituted chroman-4-one is shown as the following formula (a),
in the formula (a), R 1 The radicals are isobutyl, n-butyl, cyclopentylmethyl, cyclopropylmethyl, cyclohexylmethyl, benzyl, 4-methoxybenzyl, tert-butyldimethylsilyl, (tetrahydro-2H-pyran-4-yl) methyl, 2- (piperidin-1-yl) ethyl, 2-morpholinoethyl, 2- (methyl (phenyl) amino) ethyl or 2- (1H-imidazol-1-yl) ethyl, R 2 The group is tert-butyldimethylsilyl, isobutyl or cyclopentylmethyl.
The 4', 7-disubstituted chroman-4-ones according to the invention are preferably one of the following compounds:
when R is 1 Is n-butyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is cyclopentylmethyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is cyclopropylmethyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is different fromButyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is cyclohexylmethyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is 4-methoxybenzyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is tert-butyldimethylsilyl, R 2 When the compound is isobutyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is isobutyl, R 2 When the compound is cyclopentylmethyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is n-butyl, R 2 When the compound is cyclopentylmethyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is cyclopentylmethyl, R 2 When the compound is cyclopentylmethyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is cyclopropylmethyl, R 2 When the compound is cyclopentylmethyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is cyclohexylmethyl, R 2 When the compound is cyclopentylmethyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is as follows:
when R is 1 Is morpholinoethyl, R 2 In the case of tert-butyldimethylsilyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is:
when R is 1 Is 2- (N-methylanilino) ethyl, R 2 In the case of tert-butyldimethylsilyl, the chemical structure of the 4', 7-disubstituted chroman-4-one is:
a second object of the present invention is to provide a method for synthesizing 4', 7-disubstituted chroman-4-ones, which method (as shown in reaction formula i) comprises the steps of:
naringenin and tert-butyl dimethyl chlorosilane are catalyzed by imidazole and 4-dimethylaminopyridine to obtain a compound a and a compound b, and the compound a and R are catalyzed by triphenylphosphine and diisopropyl azodicarboxylate 2 OH to obtain a compound c, removing a TBS protecting group by TBAF to obtain a compound d, and catalyzing the compound d and R by triphenylphosphine and diisopropyl azodicarboxylate 1 OH reacts to obtain a compound e; under the catalysis of triphenylphosphine and diisopropyl azodicarboxylate, compounds b and R 1 OH reaction to give compound f, wherein R 1 The radicals are isobutyl, N-butyl, cyclopentylmethyl, cyclopropylmethyl, cyclohexylmethyl, benzyl, 4-methoxybenzyl, tert-butyldimethylsilyl, (tetrahydro-2H-pyran-4-yl) methyl, 2- (piperidin-1-yl) ethyl, 2-morpholinoethyl, 2- (N-methylanilino) ethyl or 2- (1H-imidazol-1-yl) ethyl, R 2 The group is tert-butyldimethylsilyl, isobutyl or cyclopentylmethyl;
the reaction formula of the method is as follows:
experiments show that the 4', 7-disubstituted chroman-4-one can inhibit the release of cytokines, reduce the generation of pro-inflammatory mediators and inhibit the occurrence and development of inflammatory reactions.
It is therefore a third object of the present invention to provide the use of the above 4', 7-disubstituted chroman-4-ones for the preparation of anti-inflammatory medicaments.
A fourth object of the present invention is to provide an anti-inflammatory agent comprising the above 4', 7-disubstituted chroman-4-one as an active ingredient.
Preferably, the anti-inflammatory agent is an anti-pulmonary inflammatory agent.
Further preferably, the anti-inflammatory agent is an agent for treating chronic obstructive pulmonary disease COPD.
Preferably, the anti-inflammatory agent comprises 4', 7-disubstituted chroman-4-one and pharmaceutically acceptable excipients.
The invention synthesizes 4', 7-disubstituted chroman-4-one, and experiments show that the 4', 7-disubstituted chroman-4-one can effectively inhibit inflammatory reaction of lung. Therefore, can be used for inhibiting lung inflammatory reaction and has wide application.
Description of the drawings:
FIG. 1 shows the structure of compound b as verified by nuclear magnetic single crystal diffraction;
figure 2 is a graph showing the results of anti-inflammatory activity of the compounds.
Detailed Description
The following examples are further illustrative of the invention and are not intended to be limiting thereof.
Example 1 synthesis of key intermediates of the invention and synthesis of the target product steps:
naringenin YPS (10 g,36.76 mmol) was weighed out, transferred to a 500ml round bottom flask equipped with a stirrer, and then 30ml of dichloromethane was added and stirred at room temperature for 10min. Imidazole (5 g,73.52 mmol) and 4-dimethylaminopyridine (0.9 g,7.4 mmol) were added sequentially and stirred at room temperature for 30min until the solution was clear. T-butyldimethylchlorosilane (5.6 g,37.33 mmol) was added slowly in portions, stirred overnight at room temperature, and the progress of the reaction monitored by TLC. On the next day, when the conversion rate of the reaction reached about 80% or more, the reaction was quenched by adding 30ml of saturated sodium bicarbonate solution, extracted three times with 30ml of dichloromethane, the organic phases were combined, washed with saturated saline solution, dried over anhydrous sodium sulfate, filtered, the solvent was dried by spin-drying, and passed through a column to give a yellow oily solid (intermediate a), yield: 55.8%. IR (heat, cm) -1 ):3358,2956,2932,2888,2859,1641,1569,1518,1469,1371,1345,1309,1269,1181,1089,1068,1018,836,784. 1 H NMR(600MHz,Chloroform-d)δ11.95(s,1H),7.33(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.99(dd,J＝19.2,2.4Hz,2H),5.35(dd,J＝13.2,3.0Hz,1H),5.20(s,1H),3.09(dd,J＝17.4,13.2Hz,1H),2.77(dd,J＝17.4,3.0Hz,1H),0.96(s,9H),0.24(s,6H). 13 CNMR(150MHz,Chloroform-d)δ196.2,165.0,163.8,162.8,156.1,130.5,127.9,115.6,103.5,101.2,99.8,78.8,43.2,25.4,18.1,-4.4.HRMS(CI + )m/z calculated for C 21 H 27 O 5 Si[M+H] + 387.1622,found 387.1617.
Intermediate b (structure verified by nuclear magnetic single crystal diffraction, fig. 1), off-white crystalline solid, yield: 33.4%. IR (heat, cm) -1 ):3278,2957,2925,2855,1714,1641,1512,1464,1379,1340,1269,1165,1085,915,837,808,779. 1 H NMR(600MHz,Chloroform-d)δ12.05(s,1H),7.31(d,J＝8.4Hz,2H),6.88(d,J＝9.0Hz,2H),6.41(s,1H),5.99(dd,J＝9.6,2.4Hz,2H),5.35(dd,J＝13.2,2.4Hz,1H),3.09(dd,J＝17.4,13.2Hz,1H),2.78(dd,J＝17.4,3.0Hz,1H),0.99(s,9H),0.21(s,6H). 13 C NMR(150MHz,Chloroform-d)δ196.3,164.9,164.3,163.4,156.3,130.8,127.7,120.4,103.1,96.7,95.6,79.1,43.2,25.7,18.2,-4.4.HRMS(CI + )m/z calculated for C 21 H 27 O 5 Si[M+H] + 387.1622,found387.1618.
Synthesis of intermediate c:
dissolving the compound a in tetrahydrofuran, stirring at room temperature for 10min, and sequentially adding triphenylphosphine and R 2 After OH and stirring at room temperature for 10min, dropwise adding diisopropyl azodicarboxylate into an ice bath in a nitrogen atmosphere, gradually returning to room temperature, reacting at room temperature for 3h, and performing column chromatography separation after TLC detection reaction is finished to obtain a compound c.
R 2 When OH is isobutanol, compound C (yellow powdered solid), yield: 56.0%. IR (heat, cm) -1 ):3321,2964,2917,1631,1593,1566,1519,1466,1377,1348,1295,1270,1213,1177,1096,1054,1027,883,827,735. 1 H NMR(400MHz,Chloroform-d)δ12.00(s,1H),7.33(d,J＝8.4Hz,2H),6.88(d,J＝8.4Hz,2H),6.05(dd,J＝9.2,2.0Hz,2H),5.35(dd,J＝12.8,2.8Hz,1H),5.05(s,1H),3.73(d,J＝6.4Hz,2H),3.08(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,3.2Hz,1H),2.10–2.03(m,1H),1.00(d,J＝6.4Hz,6H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,156.0,130.6,127.9,115.6,102.9,95.5,94.5,78.8,74.7,43.1,27.9,19.0.HRMS(CI + )m/z calculated for C 19 H 21 O 5 [M+H] + 329.1384,found 329.1379.
R 2 When OH is cyclopentyl methanol, compound C (yellow oily solid), yield: 58.0%. IR (heat, cm) -1 ):2955,2860,1644,1612,1574,1513,1464,1372,1341,1304,1268,1161,1091,1027,915,839,806,782,746. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.31(d,J＝8.4Hz,2H),6.88(d,J＝8.4Hz,2H),6.05(dd,J＝7.2,2.4Hz,2H),5.34(dd,J＝13.2,3.2Hz,1H),3.84(d,J＝6.8Hz,2H),3.08(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝16.8,2.8Hz,1H),2.40–2.28(m,1H),1.86–1.78(m,2H),1.64–1.58(m,4H),1.31(d,J＝19.6Hz,2H),0.99(s,9H),0.21(s,6H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,156.1,131.0,127.5,120.3,102.9,95.5,94.5,79.0,72.6,43.2,38.6,29.3,25.6,25.3,18.1,1.0.HRMS(CI + )m/z calculated for C 27 H 37 O 5 Si 1 [M+H] + 469.2405,found 469.2398.
Synthesis of intermediate d:
dissolving the compound c in dichloromethane, stirring in ice bath for 10min, dripping tetrabutylammonium fluoride, reacting in ice bath for 0.5h, and performing column chromatography separation after TLC detection reaction is finished to obtain the compound d.
R 2 When OH is isobutanol, compound d (yellow powdered solid), yield: 56.0%. IR (heat, cm) -1 ):3321,2964,2917,1631,1593,1566,1519,1466,1377,1348,1295,1270,1213,1177,1096,1054,1027,883,827,735. 1 H NMR(400MHz,Chloroform-d)δ12.00(s,1H),7.33(d,J＝8.4Hz,2H),6.88(d,J＝8.4Hz,2H),6.05(dd,J＝9.2,2.0Hz,2H),5.35(dd,J＝12.8,2.8Hz,1H),5.05(s,1H),3.73(d,J＝6.4Hz,2H),3.08(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,3.2Hz,1H),2.10–2.03(m,1H),1.00(d,J＝6.4Hz,6H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,156.0,130.6,127.9,115.6,102.9,95.5,94.5,78.8,74.7,43.1,27.9,19.0.HRMS(CI + )m/z calculated for C 19 H 21 O 5 [M+H] + 329.1384,found 329.1379.
R 2 When OH is cyclopentyl methanol, compound d (yellow powdered solid), yield: 46.8%. IR (heat, cm) -1 ):3238,2960,2869,1735,1646,1596,1519,1460,1382,1358,1307,1273,1210,1166,1087,889,833,776,743. 1 H NMR(400MHz,Chloroform-d)δ12.00(s,1H),7.33(d,J＝8.4Hz,2H),6.88(d,J＝8.8Hz,2H),6.05(dd,J＝9.6,2.4Hz,2H),5.35(dd,J＝13.2,3.2Hz,1H),5.00(s,1H),3.84(d,J＝6.8Hz,2H),3.08(dd,J＝16.8,12.8Hz,1H),2.78(dd,J＝17.2,2.8Hz,1H),2.39–2.28(m,1H),1.85–1.78(m,2H),1.65–1.60(m,4H),1.35–1.28(m,2H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.7,164.0,162.8,156.0,130.6,127.9,115.6,102.9,95.5,94.6,78.8,72.6,43.1,38.6,29.3,25.3.HRMS(CI + )m/z calculated for C 21 H 21 O 5 [M-H] - 353.1395,found 353.1400.
Synthesis of compound e:
dissolving the compound d in tetrahydrofuran, stirring at room temperature for 10min, and sequentially adding triphenylphosphine and R 1 After OH and stirring at room temperature for 10min, dropwise adding diisopropyl azodicarboxylate into an ice bath in a nitrogen atmosphere, gradually recovering the room temperature, reacting at room temperature for 4-14h, and performing column chromatography separation after TLC detection reaction is finished to obtain a target compound (compound e).
Example 2 7 Synthesis of cyclopentyloxy-5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 1):
prepared according to the method of example 1, except using the corresponding starting materials, wherein in the synthesis of intermediate c, R 2 OH is isobutanol, R in the synthesis of the target compound e 1 The OH is cyclopentanol. Yield: 66.7%. IR (heat, cm) -1 ):3053,2955,2869,1647,1617,1573,1518,1465,1427,1404,1337,1298,1254,1198,1163,1093,1073,1031,995,980,831,808,759,739. 1 H NMR(600MHz,Chloroform-d)δ12.02(s,1H),7.36(d,J＝8.4Hz,2H),6.94(d,J＝9.0Hz,2H),6.04(dd,J＝13.8,2.4Hz,2H),5.35(dd,J＝12.6,3.0Hz,1H),3.85(d,J＝6.6Hz,2H),3.72(d,J＝6.6Hz,2H),3.09(dd,J＝17.4,13.2Hz,1H),2.77(dd,J＝17.4,3.0Hz,1H),2.38–2.36(m,1H),2.09–2.05(m,1H),1.87–1.82(m,2H),1.66–1.58(m,4H),1.39–1.5(m,2H),1.00(d,J＝7.2Hz,6H). 13 C NMR(150MHz,Chloroform-d)δ196.0,167.7,164.1,162.9,159.8,130.2,127.7,114.8,95.6,94.6,79.0,74.8,72.4,43.2,39.0,29.5,28.0,25.4,19.1.HRMS(CI + )m/z calculated for C 25 H 31 O 5 [M+H] + 411.2166,found 411.2159.
Example 37 Synthesis of n-butoxy-5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 2):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is isobutanol, R in the synthesis of the target compound e 1 N-butanol is used for OH. Yield: 80.0%. IR (heat, cm) -1 ):2954,2872,1638,1585,1518,1462,1444,1376,1352,1306,1273,1209,1165,1085,1069,1028,972,888,829,796,744. 1 H NMR(400MHz,Chloroform-d)δ12.02(s,1H),7.36(d,J＝8.4Hz,2H),6.94(d,J＝8.4Hz,2H),6.04(dd,J＝9.2,2.4Hz,2H),5.34(dd,J＝12.8,2.8Hz,1H),3.98(t,J＝6.8Hz,2H),3.72(d,J＝6.4Hz,2H),3.08(dd,J＝16.8,12.8Hz,1H),2.77(dd,J＝17.2,3.2Hz,1H),2.12–2.02(m,1H),1.82–1.74(m,2H),1.55–1.46(m,2H),1.01–0.97(m,9H). 13 CNMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.5,130.1,127.6,114.6,102.9,95.4,94.5,78.9,74.7,67.7,43.1,31.2,27.9,19.1,19.0,13.8.HRMS(CI + )m/z calculated for C 23 H 27 O 5 [M-H] - 383.1864,found 383.1869.
Example 4 7 Synthesis of cyclopropylmethoxy-5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 3):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is isobutanol, R in the synthesis of the target compound e 1 The OH is cyclopropyl alcohol. Yield: 76.7%. IR (heat, cm) -1 ):2958,1632,1581,1517,1467,1445,1377,1352,1307,1280,1251,1211,1167,1084,1027,1010,886,840,803,744. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.36(d,J＝8.4Hz,2H),6.95(d,J＝8.8Hz,2H),6.04(dd,J＝9.2,2.4Hz,2H),5.35(dd,J＝12.8,2.8Hz,1H),3.82(d,J＝6.8Hz,2H),3.72(d,J＝6.4Hz,2H),3.08(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,3.2Hz,1H),2.12–2.02(m,1H),1.29–1.25(m,1H),1.00(d,J＝6.8Hz,6H),0.68–0.63(m,2H),0.38–0.34(m,2H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.4,130.3,127.6,114.7,95.5,94.5,78.9,74.7,72.8,43.1,27.9,19.0,10.1,3.2.HRMS(CI + )m/z calculated for C 23 H 27 O 5 [M+H] + 383.1853,found 383.1839.
Example 57 Synthesis of p-methoxybenzyloxy-5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 4):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is isobutanol, R in the synthesis of the target compound e 1 The OH is p-methoxybenzyl alcohol. Yield: 66.7%. IR (heat, cm) -1 ):3078,2959,2923,2857,1647,1627,1578,1516,1468,1398,1373,1349,1304,1245,1204,1165,1097,1066,1017,837,808,742. 1 H NMR(600MHz,Chloroform-d)δ12.01(s,1H),7.37(dd,J＝8.4,5.4Hz,4H),7.01(d,J＝8.4Hz,2H),6.93(d,J＝9.0Hz,2H),6.04(dd,J＝15.0,2.4Hz,2H),5.36(dd,J＝12.6,3.0Hz,1H),5.01(s,2H),3.82(s,3H),3.72(d,J＝6.6Hz,2H),3.09(dd,J＝16.8,13.2Hz,1H),2.78(dd,J＝16.8,3.0Hz,1H),2.09–2.05(m,1H),1.00(d,J＝6.6Hz,6H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.5,159.2,130.5,129.2,128.6,127.6,115.0,114.0,102.9,95.5,94.5,78.9,74.7,69.8,55.3,43.1,27.9,19.0.HRMS(CI + )m/z calculated for C 27 H 27 O 6 [M-H] - 447.1813,found 447.1823.
Example 6 7 Synthesis of (t-butyldimethylsilyl) oxy-5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 5):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 The use of isobutanol for OH eliminates the need for the latter 2-step reaction of example 1 to give compound c (i.e., compound e 5). Yield: 64.3%. IR (heat, cm) -1 ):2958,2931,2859,1644,1612,1574,1513,1469,1372,1341,1304,1268,1163,1091,916,839,806,782. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.31(d,J＝8.4Hz,2H),6.88(d,J＝8.8Hz,2H),6.05(dd,J＝6.4,2.0Hz,2H),5.35(dd,J＝12.8,2.8Hz,1H),3.73(d,J＝6.8Hz,2H),3.08(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,2.8Hz,1H),2.12–2.02(m,1H),1.00–0.99(m,15H),0.22(s,6H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,156.1,131.0,127.5,120.3,102.9,95.5,94.5,79.0,74.7,43.2,27.9,25.6,19.0,18.1,1.0.HRMS(CI + )m/z calculated for C 25 H 35 O 5 Si 1 [M+H] + 443.2248,found 443.2239.
Example 7 7 Synthesis of 5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 6):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is isobutanol, R in the synthesis of the target compound e 1 Isobutanol was used for OH. Yield: 76.7%. IR (heat, cm) -1 ):3345,2963,2937,2913,2878,1653,1614,1573,1513,1470,1403,1357,1302,1263,1245,1169,1119,1095,1069,1034,845,763,742. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.36(d,J＝8.4Hz,2H),6.94(d,J＝8.8Hz,2H),6.04(dd,J＝8.8,2.0Hz,2H),5.35(dd,J＝12.8,2.8Hz,1H),3.73(t,J＝6.4Hz,4H),3.09(dd,J＝17.2,12.8Hz,1H),2.78(dd,J＝17.2,2.8Hz,1H),2.15–2.02(m,2H),1.02(dd,J＝14.0,6.8Hz,12H). 13 C NMR(100MHz,Chloroform-d)δ196.1,167.8,164.3,163.1,159.9,130.4,127.8,115.0,103.2,95.7,94.8,79.1,74.9,74.7,43.4,28.4,28.2,19.4,19.2.HRMS(CI + )m/z calculated for C 23 H 29 O 5 [M+H] + 385.2010,found 385.2002.
Example 8 7 Synthesis of Cyclohexylmethoxy-5-hydroxy-2- (4-isobutoxyphenyl) chroman-4-one (e 7):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is isobutanol, R in the synthesis of the target compound e 1 The OH is cyclohexane methanol. Yield: 86.7%. IR (heat, cm) -1 ):2926,2856,1635,1579,1518,1461,1399,1369,1309,1253,1163,1097,1069,1025,893,836,814,795,744. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.35(d,J＝8.8Hz,2H),6.93(d,J＝8.8Hz,2H),6.04(dd,J＝9.2,2.4Hz,2H),5.35(dd,J＝12.8,3.2Hz,1H),3.75(dd,J＝18.8,6.4Hz,4H),3.09(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝16.8,2.8Hz,1H),2.10–2.04(m,1H),1.90–1.89(m,1H),1.81–1.70(m,5H),1.35–1.19(m,5H),1.00(d,J＝6.8Hz,6H). 13 CNMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.7,130.0,127.6,114.7,102.9,95.5,94.5,78.9,74.7,73.5,43.1,37.6,29.8,27.9,26.4,25.7,19.0.HRMS(CI + )m/z calculated for C 26 H 33 O 5 [M+H] + 425.2323,found 425.2319.
Example 9 7 Synthesis of n-butoxy-5-hydroxy-2- (4-cyclopentmethoxyphenyl) chroman-4-one (e 8):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is cyclopentyl methanol, R in the synthesis of target compound e 1 OH useIs n-butanol. Yield: 56.7%. IR (heat, cm) -1 ):2951,2865,1651,1616,1579,1516,1464,1376,1330,1308,1278,1257,1244,1213,1168,1089,1029,974,889,829,795,746. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.36(d,J＝8.8Hz,2H),6.94(d,J＝8.4Hz,2H),6.08(dd,J＝9.2,2.4Hz,2H),5.35(dd,J＝12.8,3.2Hz,1H),3.98(t,J＝6.4Hz,2H),3.84(d,J＝7.2Hz,2H),3.09(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,3.2Hz,1H),2.39–2.28(m,1H),1.85–1.74(m,4H),1.65–1.55(m,4H),1.53–1.45(m,2H),1.35–1.29(m,2H),0.98(t,J＝7.2Hz,3H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.5,130.1,127.6,114.7,102.9,95.5,94.5,78.9,72.6,67.7,43.1,38.6,31.2,29.3,25.3,19.2,13.8.HRMS(CI + )m/z calculated for C 25 H 29 O 5 [M-H] - 409.2021,found 409.2029.
Example 10 Synthesis of 7-isobutoxy-5-hydroxy-2- (4-cyclopentmethoxyphenyl) chroman-4-one (e 9):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is cyclopentyl methanol, R in the synthesis of target compound e 1 Isobutanol was used for OH. Yield: 46.4%. IR (heat, cm) -1 ):2957,2870,1644,1574,1515,1467,1372,1341,1302,1253,1198,1160,1091,1030,887,830,743. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.36(d,J＝8.8Hz,2H),6.94(d,J＝8.8Hz,2H),6.04(dd,J＝9.2,2.4Hz,2H),5.35(dd,J＝12.8,3.2Hz,1H),3.84(d,J＝7.2Hz,2H),3.74(d,J＝6.8Hz,2H),3.09(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,3.2Hz,1H),2.39–2.28(m,1H),2.16–2.03(m,1H),1.86–1.78(m,2H),1.66–1.55(m,4H),1.36–1.29(m,2H),1.03(d,J＝6.4Hz,6H). 13 C NMR(100MHz,Chloroform-d)δ196.1,167.9,164.3,163.1,159.9,130.4,127.8,115.0,103.2,95.7,94.8,79.1,74.7,72.8,43.4,38.9,29.5,28.4,25.5,19.4.HRMS(CI + )m/z calculated for C 25 H 31 O 5 [M+H] + 411.2166,found 411.2158.
Example 11 Synthesis of 7-cyclohexylmethoxy-5-hydroxy-2- (4-cyclopentylmethoxyphenyl) chroman-4-one (e 10):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is cyclopentyl methanol, R in the synthesis of target compound e 1 The OH is cyclohexane methanol. Yield: 75.0%. IR (heat, cm) -1 ):3047,2962,2927,2854,1632,1579,1517,1465,1368,1311,1253,1179,1096,1070,1025,893,863,837,814,745. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.35(d,J＝8.8Hz,2H),6.93(d,J＝8.8Hz,2H),6.04(dd,J＝9.6,2.4Hz,2H),5.35(dd,J＝13.2,3.2Hz,1H),3.83(d,J＝6.8Hz,2H),3.77(d,J＝6.4Hz,2H),3.09(dd,J＝17.2,12.8Hz,1H),2.77(dd,J＝17.2,2.8Hz,1H),2.39–2.28(m,1H),1.89–1.84(m,3H),1.82–1.73(m,5H),1.64–1.58(m,4H),1.35–1.22(m,5H),1.10–1.01(m,2H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.7,130.0,127.6,114.7,102.9,95.5,94.5,78.9,73.5,72.6,43.1,38.6,37.6,29.8,29.3,26.4,25.7,25.3.HRMS(CI + )m/z calculated for C 28 H 35 O 5 [M+H] + 451.2479,found 451.2486.
Example 12 Synthesis of 7-cyclopentylmethoxy-5-hydroxy-2- (4-cyclopentmethoxyphenyl) chroman-4-one (e 11):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is cyclopentyl methanol, R in the synthesis of target compound e 1 The OH is cyclopentanol. Yield: 50.0%.IR(neat,cm -1 ):3055,2955,2865,1648,1616,1572,1517,1464,1403,1344,1301,1251,1167,1123,1093,1073,1029,989,939,864,833,810,766,744. 1 H NMR(600MHz,Chloroform-d)δ12.02(s,1H),7.36(d,J＝8.4Hz,2H),6.94(d,J＝9.0Hz,2H),6.04(dd,J＝13.8,1.8Hz,2H),5.35(dd,J＝13.2,3.0Hz,1H),3.84(t,J＝7.2Hz,4H),3.09(dd,J＝17.4,13.2Hz,1H),2.77(dd,J＝16.8,3.0Hz,1H),2.41–2.30(m,2H),1.87–1.79(m,4H),1.68–1.56(m,8H),1.39–1.28(m,4H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.7,130.1,127.6,114.7,102.9,95.5,94.5,78.90,72.6,72.3,43.1,38.9,38.6,29.3(d,J＝16.4Hz),25.3(d,J＝10.3Hz).HRMS(CI + )m/z calculated for C 27 H 33 O 5 [M+H] + 437.2323,found 437.2309.
Example 13 Synthesis of 7-cyclopropylmethoxy-5-hydroxy-2- (4-cyclopentylmethoxyphenyl) chroman-4-one (e 12):
prepared as in example 1 except using the corresponding starting materials. Wherein R in the synthesis of intermediate c 2 OH is cyclopentyl methanol, R in the synthesis of target compound e 1 The OH is cyclopropyl methanol. Yield: 82.1%. IR (heat, cm) -1 ):3088,3009,2947,2865,1650,1613,1581,1515,1445,1404,1377,1330,1307,1280,1242,1215,1166,1089,1028,890,828,801,746. 1 H NMR(400MHz,Chloroform-d)δ12.00(s,1H),7.36(d,J＝8.7Hz,2H),6.94(d,J＝8.8Hz,2H),6.04(dd,J＝9.2,2.0Hz,2H),5.35(dd,J＝13.2,3.2Hz,1H),3.83(dd,J＝7.2,4.8Hz,4H),3.08(dd,J＝17.2,13.2Hz,1H),2.78(dd,J＝17.2,2.8Hz,1H),2.40–2.28(m,1H),1.85–1.78(m,2H),1.65–1.56(m,4H),1.34–1.26(m,3H),0.68–0.63(m,2H),0.36(dd,J＝10.8,4.8Hz,2H). 13 C NMR(150MHz,Chloroform-d)δ195.9,167.6,164.0,162.8,159.4,130.3,127.6,114.7,102.9,95.5,94.5,78.9,72.8,72.6,43.1,38.6,29.3,25.3,10.1,3.2.HRMS(CI + )m/z calculated for C 25 H 29 O 5 [M+H] + 409.2010,found 409.1998.
Example 14 synthesis of 7-morpholinoethoxy-5-hydroxy-2- (4- (tert-butyldimethylsilyl) oxyphenyl) chroman-4-one (e 13):
dissolving the compound b in tetrahydrofuran, stirring at room temperature for 10min, and sequentially adding triphenylphosphine and R 1 After OH and stirring at room temperature for 10min, dropwise adding diisopropyl azodicarboxylate into an ice bath in a nitrogen atmosphere, gradually recovering the room temperature, reacting at room temperature for 3h, and performing column chromatography separation after TLC detection reaction is finished to obtain a compound f.
When R is 1 When morpholine ethanol is used as OH, the yield of compound e13 (i.e., compound f 1): 61.7%. IR (heat, cm) -1 ):3300,2960,2928,2855,1726,1638,1573,1512,1543,1454,1374,1342,1301,1267,1199,1165,1119,1089,1031,917,839,803,110. 1 H NMR(400MHz,Chloroform-d)δ12.01(s,1H),7.30(d,J＝8.4Hz,2H),6.88(d,J＝8.0Hz,2H),6.05(d,J＝6.0Hz,2H),5.35(d,J＝10.4Hz,1H),4.11(t,J＝5.6Hz,2H),3.71(d,J＝4.8Hz,4H),3.08(dd,J＝16.8,12.8Hz,1H),2.79(d,J＝6.0Hz,2H),2.77(s,1H),2.56(s,4H),0.99(s,9H),0.21(s,6H). 13 C NMR(125MHz,Chloroform-d)δ196.0,166.8,164.0,162.8,156.1,130.8,127.5,120.3,103.1,95.5,94.5,79.0,66.7,66.2,57.1,53.9,43.1,29.6,25.5,-4.5.HRMS(CI + )m/z calculated for C 27 H 38 NO 6 Si[M+H] + 500.2463,found500.2458.
Example 15 Synthesis of 7- (N-methyl (phenyl) amino) ethoxy-5-hydroxy-2- (4- (tert-butyldimethylsilyl) oxyphenyl) chroman-4-one (e 14):
prepared as in example 14 except using the corresponding starting materials. Wherein R is 1 The OH group is 2- (N-methylanilino) ethanol. Yield of compound e14 (i.e., compound f 2): 36.7%. IR (heat, cm) -1 ):3513,3292,2923,2848,1631,1568,1507,1444,1374,1349,1293,1260,1194,1172,1077,1036,822,800,742,686. 1 H NMR 1 H NMR(400MHz,Chloroform-d)δ11.99(s,1H),7.31(d,J＝8.0Hz,2H),7.23(d,J＝7.2Hz,2H),6.88(d,J＝8.0Hz,2H),6.73(t,J＝8.0Hz,3H),6.02(d,J＝9.6Hz,2H),5.34(d,J＝10.8Hz,1H),4.14(t,J＝5.6Hz,2H),3.74(t,J＝5.6Hz,2H),3.07(dd,J＝17.2,4.0Hz,1H),3.02(s,3H),2.78(dd,J＝17.2,3.2Hz,1H),0.88(s,1H). 13 C NMR(125MHz,Chloroform-d)δ195.9,166.9,164.0,162.8,156.1,148.6,130.4,129.2,127.8,116.7,115.6,112.1,103.1,95.4,94.5,78.8,65.8,51.5,43.1,39.0.HRMS(CI + )m/z calculated for C 24 H 24 NO 5 [M+H] + 406.1649,found406.1645.
EXAMPLE 16 investigation of 4', 7-disubstituted chroman-4-ones for anti-inflammatory Activity
The derivatives of the invention are subjected to inhibition test of the expression level of the pneumonia cytokines, and the conventional QPCR method is adopted as the test method.
1. Cell culture
Human lung epithelial cells BEAS-2B were cultured in DMEM high-glucose medium (complete medium) supplemented with 10% (V/V) FBS, 100U/mL penicillin and 100. Mu.g/mL streptomycin. The culture conditions were 37℃and 5% CO 2 Cells were passaged to 80% -90% confluency.
2. Cell administration and induction of cellular inflammation
Experiments were performed with cells in log phase of growth, 1 x 10 cells 5 Inoculating the cells/well into 24-well plate at 37deg.C with 5% CO 2 Is cultivated until 90 percent of the culture medium is grown for standby. Setting groups: blank stimulation group (|), LPS stimulation group (+), positive control group (naringenin) and dosing group (e 1-14). Carefully remove the medium, positive control and dosing groupsFresh complete medium containing 100. Mu.M compound was added separately, and equal volumes of DMSO were added to the blank and LPS stimulated groups. After 1 hour, 1. Mu.g/mL LPS was added separately to induce cellular inflammation for 2 hours except for the blank stimulation group.
3. RNA extraction and qPCR measurement
1. After the cultured cells are completely discarded from the culture medium, 0.5mL of RA2 in the RNA extraction kit is added into each hole for lysis (the cells can be frozen at-80 ℃ for one night, the lysis effect is better, and the cells can be collected after being lysed at 4 ℃ for 10 min);
2. collecting cell lysate, extracting Total RNA according to the instruction of the kit, and measuring the concentration of Total RNA by using an ultra-micro ultraviolet visible spectrophotometer;
RT-PCR (20 uL per system) involving 5 XBUFFER 4 uL, total RNAX uL (1 pg-1ug, 200-400ng most of the time), DEPC-H 2 Placing O (16-X) mu L in an 8-connecting tube, marking serial numbers, placing the O in a PCR instrument, and performing reverse transcription at 50 ℃ for 15min,85 ℃ for 5s and 16 ℃ for a period of time;
4. 80. Mu.L DEPC-H was added to the transcribed cDNA 2 Centrifuging and mixing uniformly until the concentration of O is 100 mu L, and keeping the temperature at-20 ℃ for later use;
5. according to (SYBR 10. Mu.L+DEPC-H) 2 O10 μL+0.5 μL of primer) multiplied by the number of samples, 18 μL/well was added to a 96-well plate, and 2 μL cDNA was added to form a 20 μL system, and the mixture was centrifuged at 1200rpm for 1min to mix well;
6. the sample plate is put into a CFX Connect Real-Time System (Real-Time fluorescence quantitative PCR instrument) for detection, and the detection is carried out according to the procedures of (95 ℃ 2min,95 ℃ 20s,57 ℃ 20s,72 ℃ 20 s) total circulation 39 times, 95 ℃ 1min,55 ℃ 30s and 95 ℃ 30s, and the total Time of the instrument is 2.5h;
after 7.2 h, the data were saved and analyzed using 2 -ΔΔCt The experimental result is analyzed by the method, and the calculation formula is as follows: delta Ct target gene=ct target gene-Ct reference gene, delta Ct target gene= delta Ct experimental group target gene-delta Ct control group target gene. 2 -ΔΔCt Expression fold of the target gene of the experimental group relative to the control group is shown.
Design of amplification primers: the invention aims at IL-6 gene, and takes GAPDH as reference gene. Searching target genes in a gene library in NCBI, checking target gene sequences in GeneBank, and designing primer parameters according to detection requirements as shown in the following table 1.
TABLE 1
Synthesis of cDNA: cDNA is synthesized by taking total RNA as a template, 5x HiScript II Q Select RT SuperMix is adopted, each solution is uniformly mixed by vortex oscillation before use, and liquid remained on the pipe wall is collected after rapid centrifugation. After completion of the preparation of the reaction system in an ice bath and thawing of the template RNA on ice, a reverse transcription reaction system was prepared as shown in Table 2:
TABLE 2
The genome removal and reverse transcription reactions were performed in a Veriti 96well Thermal Cycler PCR apparatus, the reaction procedure being: 15min at 50 ℃ and 5s at 85 ℃, and placing the reverse transcription product in a refrigerator at-20 ℃ for standby.
Real-time qPCR: the real-time qPCR reaction system was prepared as shown in Table 3, and the primer sequences used in this experiment are shown in Table 1, and GAPDH gene was selected as an internal reference, and the primers were synthesized by the division of biological engineering (Shanghai).
TABLE 3 Table 3
Real-Time qPCR reaction is carried out in a CFX Connect Real-Time System Real-Time fluorescent quantitative PCR instrument, and the amplification procedure is as follows: 2min at 95℃and then 40 cycles: 95℃20s,57℃20s,72℃20s. The temperature is increased from 55 ℃ to 95 ℃ to obtainTo a melting curve. The results are automatically analyzed by analysis software to generate an amplification curve and calculate Ct values. By 2 -ΔΔct The experimental result is analyzed by the method, and the calculation formula is as follows: delta Ct Target gene ＝Ct Target gene -Ct Reference gene ，△△Ct Target gene ＝△Ct Target gene of experimental group -△Ct Target gene of control group 。2 -ΔΔct Expression fold of target gene of the experimental group relative to the control group is shown, and normalization treatment is carried out by taking LPS stimulated group as a reference.
The results are shown in Table 4:
TABLE 4 Table 4
The test results show that 4', 7-disubstituted chroman-4-ones are effective in inhibiting IL-6 expression levels in the tested human lung epithelial cell lines (FIG. 2), and that compounds e8, e9, e11 and e14 exhibit better anti-inflammatory activity. The experimental result shows that the compound has good anti-pneumonia activity and can be used for researching anti-lung inflammation medicines.

Claims (9)

1.4', 7-disubstituted chroman-4-ones, wherein the 4', 7-disubstituted chroman-4-ones are any of the following:
、
、
、
。
2. a method for synthesizing 4', 7-disubstituted chroman-4-ones according to claim 1, wherein the method is represented by formula i, comprising the steps of:
naringenin and tert-butyl dimethyl chlorosilane are catalyzed by imidazole and 4-dimethylaminopyridine to obtain a compound a and a compound b, and the compound a and R are catalyzed by triphenylphosphine and diisopropyl azodicarboxylate 2 OH to obtain a compound c, removing a TBS protecting group by TBAF to obtain a compound d, and catalyzing the compound d and R by triphenylphosphine and diisopropyl azodicarboxylate 1 OH reacts to obtain a compound e; under the catalysis of triphenylphosphine and diisopropyl azodicarboxylate, compounds b and R 1 OH reaction to give compound f, wherein R 1 The radical being isobutyl, N-butyl, cyclopentylmethyl, 2- (N-methylanilino) ethyl, R 2 The group is tert-butyldimethylsilyl or cyclopentylmethyl;
the reaction formula of the method is as follows:
(i)。
3. use of the 4', 7-disubstituted chroman-4-ones of claim 1 for the preparation of anti-inflammatory medicaments.
4. The use according to claim 3, wherein the anti-inflammatory agent is an anti-pulmonary inflammatory agent.
5. The use according to claim 4, wherein the anti-inflammatory agent is an agent for the treatment of chronic obstructive pulmonary disease COPD.
6. An anti-inflammatory agent comprising the 4', 7-disubstituted chroman-4-one of claim 1 as an active ingredient.
7. The anti-inflammatory agent according to claim 6, wherein the anti-inflammatory agent is an anti-pulmonary inflammatory agent.
8. The anti-inflammatory agent according to claim 7, wherein the anti-inflammatory agent is an agent for treating chronic obstructive pulmonary disease COPD.
9. The anti-inflammatory agent according to claim 6, wherein the anti-inflammatory agent comprises the 4', 7-disubstituted chroman-4-one of claim 1 and a pharmaceutically acceptable adjuvant.
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