CN113620943B - Nitrofuran compound, pharmaceutical composition, preparation method and application thereof - Google Patents
Nitrofuran compound, pharmaceutical composition, preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a nitrofuran compound, a pharmaceutical composition, a preparation method and application thereof. The nitrofuran compound has a structure shown as a formula (I):
the nitrofuran compound provided by the invention has a skeleton with the advantage of drug property, has the characteristics of drug forming property and the like, has high selectivity on STAT3 protein, has a remarkable inhibition effect on STAT3, and can be used as a STAT3 specific inhibitor; in addition, the nitrofuran compound can obviously reduce the volume and weight of tumors in vivo and obviously inhibit the growth and proliferation of the tumors; in addition, when the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof and the EGFR inhibitor are used together, the generation of acquired drug resistance of the EGFR drug can be delayed, the clinical service life of the EGFR drug can be prolonged, and the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof has important clinical significance.
Description
Technical Field
The invention relates to the field of pharmaceutical chemistry and pharmacotherapeutics, in particular to a nitrofuran compound, a pharmaceutical composition, a preparation method and application thereof.
Background
The transcription factor STAT3 is a bifunctional protein, has dual functions of signal transduction and transcriptional activation, and is responsible for regulating and controlling a series of important physiological processes of cell growth, proliferation, differentiation, apoptosis and the like. However, studies find that STAT3 has persistent activation and abnormally high expression, can induce cell proliferation, invasion and migration, inhibit apoptosis, promote angiogenesis, and play an important role in the process of tumor generation, development and metastasis. Therefore, the development of targeted STAT3 inhibitors has been a hot spot in the frontier of antitumor drug research.
Currently, several targeted STAT3 drug candidates have entered phase I/II clinical trials, such as: s31-201, STA-21, Niclosamide, C188-9, OPB-31121, BP-1-102 and pyrimethanamine, but no drug is yet approved for clinical treatment. Where most compounds under investigation were non-specific STAT3 inhibitors, lacking selectivity; on the other hand, the proliferation inhibition effect of most compounds on STAT3 high-expression tumor cell lines is still at micromolar level; moreover, many compounds also face the problem of drug forming properties such as low oral bioavailability (e.g. Niclosamide), poor solubility (e.g. C188-9) and unstable structure (e.g. BP-1-102), and further structural optimization is still needed to improve drug forming properties.
In conclusion, specific inhibitors targeting STAT3 have been the focus of research and development (e.g., European Journal of Medicinal Chemistry,2020,187,111922), but the drugs developed so far all have the disadvantages of small drug effect, low selectivity, poor drug potency, and the like, in different degrees and different aspects, and limit the clinical application and later development of STAT3 inhibitors. While STAT3 is a promising tumor treatment target, the development of a new class of compounds which can specifically inhibit STAT3 and have the characteristics of high selectivity, strong drug effect, good drug property and the like is urgently needed in the field.
Disclosure of Invention
The invention aims to overcome the defects or shortcomings of small efficacy, low selectivity, poor drug property and the like of the existing STAT3 and provides a nitrofuran compound. The nitrofuran compound provided by the invention has a skeleton with the advantage of drug property, has the characteristics of drug forming property and the like, has high selectivity on STAT3 protein, has a remarkable inhibition effect on STAT3, and can be used as a STAT3 specific inhibitor; in addition, the nitrofuran compound can obviously reduce the volume and weight of tumors in vivo and obviously inhibit the growth and proliferation of the tumors; in addition, when the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof and the EGFR inhibitor are used together, the generation of acquired drug resistance of the EGFR drug can be delayed, the clinical service life of the EGFR drug can be prolonged, and the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof has important clinical significance.
The invention also aims to provide a preparation method of the nitrofuran compound.
The invention also aims to provide application of the nitrofurans or pharmaceutically acceptable salts or solvates thereof in preparing a medicament for inhibiting STAT3 protein activity.
The invention also aims to provide the application of the nitrofurans compound or the pharmaceutically acceptable salt or solvate thereof in preparing the medicines for inhibiting the growth and proliferation of tumors
It is another object of the present invention to provide pharmaceutical compositions.
In order to achieve the above purpose of the present invention, the present invention provides the following technical solutions:
the nitrofuran compound has a structure shown as a formula (I):
wherein:
R 1 、R 2 、R 3 、R 4 independently selected from hydrogen, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Alkylamino, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 3-8 Cycloalkoxy, substituted or unsubstituted C 3-8 Cycloalkylamino group, substituted or unsubstituted aryl group, substituted or unsubstituted heteroaryl group, substituted or unsubstituted 3-to 8-membered heterocyclic group containing 1 to 2 hetero atoms selected from N and O、-COR a 、-CO 2 R a 、-CONR a R b 、-NR a C(O)R b 、-NR a SO 2 R b 、-SR a 、-SOR a 、-SO 2 R a 、-SO 2 NR a R b 、-OC(O)R a or-OC (O) NR a R b ，
R a And R b Independently of one another is hydrogen, C 1-6 Alkyl radical, C 3-6 Cycloalkyl, aryl or heteroaryl;
R 5 is hydrogen, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, substituted or unsubstituted C 1-8 Alkyl, substituted or unsubstituted C 1-8 Alkoxy, substituted or unsubstituted C 1-8 Alkylamino, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or substituted C 3-8 Cycloalkoxy, substituted or unsubstituted C 1-8 Alkylamino, substituted or unsubstituted aryl or heteroaryl;
R 6 、R 7 、R 8 、R 9 independently selected from hydrogen, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, C 1-3 Alkoxy radical, C 1-3 An alkylamino group.
Through repeated research, the inventor of the invention finds that the obtained nitrofuran compound has the characteristics of drug forming property and the like, has high selectivity on STAT3 protein, has a remarkable inhibition effect on STAT3, can be used as a STAT3 specific inhibitor, and has the characteristics of high selectivity, strong drug effect, good drug forming property, safety and the like for inhibiting STAT3 protein by constructing a skeleton with the advantage of drug property and reasonably substituting the skeleton.
In addition, the nitrofuran compound can obviously reduce the volume and weight of tumors in vivo and obviously inhibit the growth and proliferation of the tumors; in addition, when the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof and the EGFR inhibitor are used together, the generation of acquired drug resistance of the EGFR drug can be delayed, the clinical service life of the EGFR drug can be prolonged, and the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof has important clinical significance.
Preferably, R 1 、R 2 、R 3 、R 4 Is substituted at least at 1 position with the following substituents: halogen, cyano, amino, nitro, hydroxy, trifluoromethyl, C 1-3 Alkyl radical, C 1-3 Alkoxy radical, C 1-3 Alkylamino, 3-to 8-membered heterocyclyl containing 1-2 heteroatoms selected from N and O, -COR a ’、-CO 2 R a ’、-CONR a ’R b ’、-NR a ’C(O)R b ’、-NR a ’SO 2 R b ’、-SR a ’、-SOR a ’、-SO 2 R a ’、-SO 2 NR a ’R b ’、-OC(O)R a ’、-OC(O)NR a ’R b ’。
R a ' and R b ' independently is hydrogen, C 1-6 Alkyl radical, C 3-6 Cycloalkyl, aryl or heteroaryl;
preferably, R 1 、R 2 、R 3 、R 4 Independently selected from H, halogen, cyano, nitro, amino, hydroxyl, trifluoromethyl, C 1-6 Alkyl radical, C 1-6 Heteroalkylalkoxy, C 1-6 Alkylamino radical, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkoxy, C 3-8 Cycloalkylamino, aryl, heteroaryl, 3-to 8-membered heterocyclyl containing 1-4 heteroatoms selected from N and O.
More preferably, the 3-to 8-membered heterocyclic group containing 1 to 4 heteroatoms selected from N and O is a group including, but not limited to: 1- (3-oxetanyl) piperazinyl, homopiperazinyl, N-methylpiperazinyl, 1-t-butoxycarbonylpiperazinyl, morpholinyl, 1-thiomorpholinyl or 3, 6-dihydropyridinyl.
Preferably, R 5 Is substituted at least at 1 position with the following substituents: halogen, cyano, amino, nitro, hydroxy, trifluoromethyl, methylthio, C 1-3 Alkyl radical, C 1-3 Alkoxy or C 1-3 An alkylamino group.
Preferably, R 1 、R 2 、R 3 、R 4 、R 5 Wherein said aryl or heteroaryl is independently selected from the group consisting of:furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, benzofuryl, benzothienyl, benzoxazolyl, benzothiazolyl, phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, quinolinyl, or naphthyl.
Preferably, the nitrofurans compound has a structure shown in the following number 1-26:
the preparation method of the nitrofuran compound comprises the following steps:
s1: dissolving the formula (1) and the formula (2) in a solvent, and generating an intermediate (3) after reaction;
s2: inserting a side chain into the intermediate (3) to generate an intermediate (4);
s3: reducing the intermediate (4) to generate an intermediate (5), and then carrying out condensation reaction to obtain the nitrofuran compound;
specifically, the reaction process is as follows:
preferably, the solvent in S1 is one or more of acetone, methanol, ethanol, or acetonitrile.
Preferably, the temperature of the reaction in S2 is 40-100 ℃.
Preferably, in S3, side chain insertion into intermediate (3) under the catalysis of palladium catalyst generates intermediate (4).
The application of the nitrofurans or the pharmaceutically acceptable salt or solvate thereof in preparing the medicines for inhibiting the STAT3 protein activity is also within the protection scope of the invention.
Preferably, the pharmaceutically acceptable salt is a pharmaceutically acceptable organic or inorganic salt.
Specifically, pharmaceutically acceptable salts include, but are not limited to: sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, gluconate, formate, benzoate, glutamate, methanesulfonate (methanesulfonate), ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate; or ammonium salts (e.g., primary amine salts, secondary amine salts, tertiary amine salts, quaternary ammonium salts), metal salts (e.g., sodium salts, potassium salts, calcium salts, magnesium salts, manganese salts, iron salts, zinc salts, copper salts, lithium salts, aluminum salts).
Preferably, the nitrofurans compound or the pharmaceutically acceptable salt or solvate thereof is used for preparing a medicament for treating diseases related to abnormal cell proliferation, morphological change and hyperkinesia of STAT3 high expression, or treating diseases related to angiogenesis or cancer metastasis; in particular to the application in preparing the medicine for treating or preventing the growth and the metastasis of the tumor.
Experimental research shows that the nitrofurans compound or pharmaceutically acceptable salts and solvates thereof can obviously inhibit proliferation, migration and invasion of various in vitro tumor cells.
More preferably, the nitrofurans compound or the pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition is used for preparing the drugs for inhibiting the proliferation, migration and invasion of tumor cells.
Further preferably, the nitrofurans compound or the pharmaceutically acceptable salt or solvate thereof is used for preparing the medicine for promoting the apoptosis of tumor cells.
Preferably, cancers include, but are not limited to: acute lymphocytic leukemia, acute myelocytic leukemia, adrenocortical carcinoma, AIDS-related cancer, AIDS-related lymphoma, anal cancer, extrahepatic-biliary cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, bronchial adenoma, Burkitt's lymphoma, carcinoid tumor, cancer of unknown primary origin, central nervous system lymphoma, cervical cancer, gastric cancer, renal cancer, laryngeal cancer, blood cancer, liver cancer, non-small cell lung cancer, melanoma, prostate tumor, rectal cancer, salivary gland carcinoma, sarcoma, small intestine cancer, soft tissue sarcoma, uterine sarcoma, testicular cancer, breast cancer, ovarian cancer, rhabdoid tumor, synovial sarcoma, mesothelioma, skin cancer, oral cancer, fallopian tube tumor, peritoneal tumor, glioma, glioblastoma, myeloma.
Preferably, the tumor cell is a breast cancer cell, a lung cancer cell, a gastric adenocarcinoma and/or a gastric cancer cell.
The inventor of the invention researches a STAT3 inhibition mechanism of a nitrofuran compound through a series of experiments, and the results show that the nitrofuran compound can obviously inhibit STAT3 dimerization and combination of STAT3 and DNA, inhibit tyrosine phosphorylation level of STAT3, inhibit expression of downstream target genes BCL-XL, C-myc and Mcl-1 of STAT3, and has concentration dependence.
Preferably, the nitrofurans compound or the pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition is used for preparing the medicine for inhibiting the tyrosine phosphorylation level of STAT 3.
The invention also claims the application of the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof in preparing the medicine for inhibiting the growth and proliferation of the tumor.
The influence of the compound on in-vivo tumors is investigated through in-vivo mouse experiments, and the results show that the compound has no toxicity on visceral organs of mice, and the compound can obviously reduce the volume and weight of in-vivo tumors, namely obviously inhibit the growth and proliferation of the tumors.
The invention also claims a pharmaceutical composition, which comprises the nitrofuran compound or the pharmaceutically acceptable salt or solvate thereof and an EGFR inhibitor.
Preferably, the EGFR inhibitor is one or more of gefitinib, erlotinib hydrochloride, camitine (erlotinib hydrochloride), afatinib maleate, dacatinib, oxitinib mesylate or amitinib mesylate tablets.
Compared with the prior art, the invention has the following advantages and effects:
the nitrofuran compound provided by the invention has a skeleton with the advantage of drug property, has the characteristics of drug forming property and the like, has high selectivity on STAT3 protein, has a remarkable inhibition effect on STAT3, and can be used as a STAT3 specific inhibitor; in addition, the nitrofuran compound can obviously reduce the volume and weight of tumors in vivo and obviously inhibit the growth and proliferation of the tumors;
drawings
FIG. 1 is a plot of the growth inhibition of Compound 23 against a variety of cancer cells, as well as IC 50;
figure 2 is a graph of the results of compound 23 inhibiting phosphorylation of STAT3 and dimer formation; FIG (2A) shows the effect of compound 23 treatment on the pY705-STAT3 protein of gastric cancer cells at various time points; FIG (2B) shows the effect of different concentrations of Compound 23 on the pY705-STAT3 protein from gastric cancer cells;
FIG. 3 is a graph showing the results of inhibition of STAT3 transcriptional activity and target gene expression by Compound 23; figure (3A) shows the effect of compound 23 on STAT3 transcriptional activity; figure (3B) shows the effect of compound 23 on target genes downstream of STAT 3.
FIG. 4 is a graph showing the results of Compound 23 in inhibiting gastric cancer tumor growth in vivo; FIG. 4A is a graph showing subcutaneous gastric cancer tumors obtained by sacrifice of mice 21 days after administration to each group of nude mice; FIG. 4B is a graph showing a statistical graph of tumor volumes of nude mice during 21 days of administration to each group of nude mice; FIG. 4C is a graph showing subcutaneous gastric cancer tumor weight statistics obtained by sacrifice of mice 21 days after administration to each group of nude mice; FIG. 4D is a graph showing a statistical graph of body weights of nude mice during administration to the nude mice of each group for 21 days;
FIG. 5 is a graph showing the results of compound 23 inhibiting the expression of pY705-STAT3 protein and the target gene in vivo; FIG. 5A shows that the tissue protein of the tumor is extracted and the protein expression of pY705-STAT3 and its downstream target gene is detected after the tumor is stripped from each group of nude mice; FIG. 5B is a graph showing the results of subcutaneous gastric cancer HE staining obtained by obtaining materials from sacrificed mice 21 days after administration to each group of nude mice; FIG. 5C is a graph showing the results of HE staining of heart, liver, spleen, lung and kidney obtained by sacrifice of mice 21 days after administration to each group of nude mice.
Detailed Description
The present invention will be further explained with reference to the following examples and drawings, but the examples are not intended to limit the present invention in any manner. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the present invention are commercially available.
Example 1
The structure and preparation process of (N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (1) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 7-methoxyimidazo [1,2-a ] pyridine (1a)
4-methoxypyridin-2-amine (7.37g, 59.4mol) and 2-chloroacetaldehyde (11.64g, 148.6mol) were added to a clean flask, dissolved in 25mL of ethanol and sodium bicarbonate (9.98g, 118.9mol) was added. After stirring at 78 ℃ for 4h at rt, the fractions were extracted with ethyl acetate and water, the organic phases were combined, dried over anhydrous sodium sulfate, the solvent was dried and column chromatographed (dichloromethane/methanol 30:1, V/V) to give the desired product as a brown oil (9.95g, 87.0%). 1 H NMR(400MHz,Chloroform-d)δ7.91(d,J＝7.6Hz,1H),7.47(d,J＝1.4Hz,1H),7.39(d,J＝1.9Hz,1H),6.86(d,J＝2.5Hz,1H),6.49(dd,J＝7.4,2.5Hz,1H),3.84(s,3H).MS(EI)m/z 149.01(M+H) + .
Step 2: preparation of 7-methoxy-3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (1 b).
1a (700mg, 4.6mmol), potassium carbonate (3.174mg, 23mmol) and 1-bromo-4-nitrobenzene (1.4g, 7mmol) were charged to a clean flask at room temperature, dissolved in 10mL of N, N-dimethylformamide and the catalyst palladium acetate (51.7mg, 0.23mmol) was added to the reaction under nitrogen. After stirring at 130 ℃ for 12h, the solution was diluted with 100ml of ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The resulting solution was spun dry and purified by silica gel column chromatography (dichloromethane/methanol 65:1, V/V) to give the desired product (825mg, 67%) as a red-brown solid. 1 H NMR(500MHz,Chloroform-d)δ8.36(m,2H),8.24(d,J＝7.4Hz,1H),7.71(m,3H),6.97(d,J＝2.6Hz,1H),6.65(dd,J＝7.3,2.9Hz,1H). 13 C NMR(126MHz,Chloroform-d)δ158.37,149.20,146.47,136.09,134.52,126.94,124.78,123.84,122.95,108.62,95.76,55.70；MS(EI)m/z 270.08(M+H) + .
And step 3: preparation of 4- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) aniline (1c)
1b (690mg, 2.57mmol) was dissolved in 15mL ethanol, ammonium chloride (1374mg, 25.7mmol) was dissolved in 15mL water at room temperature, the two were mixed, and iron powder (1000mg, 17.99mmol) was added thereto. After stirring at 80 ℃ for 4 hours, the solution was filtered through celite to remove iron powder. The resulting solution was evaporated and purified by silica gel column chromatography (dichloromethane/methanol 200:1, V/V) to give the target product as a tan solid (457mg, 74%). 1 H NMR(400MHz,Chloroform-d)δ8.06(d,J＝8.2Hz,1H),7.44(s,1H),7.32–7.28(m,2H),6.91(d,J＝2.5Hz,1H),6.83–6.78(m,2H),6.51(dd,J＝7.6,2.5Hz,1H),3.88(s,3H)；MS(EI)m/z 240.11(M+H) + .
And 4, step 4: preparation of (N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (1)
Dissolve 1c (150mg, 0.62mmol) in 15mL of N, N-dimethylformamide and dissolve 5-Nitro at room temperatureTo this was added benzofuran-2-carboxylic acid (147.8mg, 0.94mmol), HATU (509.2mg, 1.34mmol) and N, N-diisopropylethylamine (239.94mg, 1.84 mmol). After stirring at room temperature for 4 hours, the solution was diluted with 100ml of ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The resulting solution was evaporated and purified by silica gel column chromatography (dichloromethane: methanol 75:1, V/V) to give the title product as a tan solid (200mg, 85%); 1 H NMR(400MHz,DMSO-d 6 )δ10.88(s,1H),8.60(d,J＝7.6Hz,1H),8.02(s,1H),8.00–7.95(m,2H),7.85(d,J＝3.9Hz,1H),7.74–7.68(m,3H),7.23(d,J＝2.6Hz,1H),7.01(dd,J＝7.6,2.6Hz,1H),3.98(s,3H).MS(EI)m/z 379.10(M+H) + ；HRMS(ESI)calcd for C 19 H 14 N 4 O 5 (M+H) + :379.1037；found 379.1035.
example 2
The structure and preparation process of N- (3-fluoro-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (2) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 3- (2-fluoro-4-nitrophenyl) -7-methoxyimidazo [1,2-a ] pyridine (2b)
The 1-bromo-4-nitrobenzene was replaced with 1-bromo-2-fluoro-4-nitrobenzene and the remaining required starting materials, reagents and preparation were the same as in step 2 of example 1 to give a tan solid.
1 H NMR(400MHz,Chlooform-d)δ8.19(dd,J＝8.5,2.3Hz,1H),8.14(dd,J＝10.0,2.3Hz,1H),7.94(dd,J＝7.5,3.2Hz,1H),7.80–7.69(m,2H),7.02(d,J＝2.5Hz,1H),6.68(dd,J＝7.6,2.5Hz,1H),3.93(s,3H)；MS(EI)m/z 288.07(M+H) + .
Step 2: 3-fluoro-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) aniline (2c)
The 1b was replaced with 2b and the remaining required starting materials, reagents and preparation were the same as in example 1, step 3, giving a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ7.91(dd,J＝7.5,2.8Hz,1H),7.38(s,1H),7.15(t,J＝8.6Hz,1H),6.99(d,J＝2.5Hz,1H),6.63(dd,J＝7.5,2.5Hz,1H),6.55–6.47(m,2H),5.74(s,2H),3.85(s,3H). 13 C NMR(126MHz,DMSO-d 6 )δ161.91,159.98,157.49,151.96,132.31,132.06,125.80,119.85,110.83,107.30,102.83,100.76,95.24,56.08；MS(EI)m/z 2548.10(M+H) + .
And step 3: preparation of N- (3-fluoro-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (2).
The 1c was replaced with 2c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ11.04(s,1H),8.27(dd,J＝7.6,2.8Hz,1H),7.95(dd,J＝12.7,2.0Hz,1H),7.89–7.83(m,2H),7.76(dd,J＝8.5,2.0Hz,1H),7.72–7.65(m,2H),7.17(d,J＝2.5Hz,1H),6.89(dd,J＝7.5,2.5Hz,1H),3.94(s,3H)；MS(EI)m/z 397.09(M+H) + ；HRMS(ESI)calcd for C 19 H 13 N 4 O 5 F(M+H) + :397.0943；found 397.0941.
Example 3
The structure and preparation process of N- (3-fluoro-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (3) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 7-methoxy-3- (2-methyl-4-nitrophenyl) imidazo [1,2-a ] pyridine (3b)
The 1-bromo-4-nitrobenzene was replaced with 1-bromo-2-methyl-4-nitrobenzene and the remaining required starting materials, reagents and preparation were the same as in step 2 of example 1 to give a tan solid.
1 H NMR(400MHz,Chloroform-d)δ8.25(d,J＝2.4Hz,1H),8.16(dd,J＝8.4,2.4Hz,1H),7.64(d,J＝7.4Hz,1H),7.54(m,2H),6.97(d,J＝2.5Hz,1H),6.58(dd,J＝7.5,2.5Hz,1H),3.90(s,3H),2.36(s,3H)；MS(EI)m/z 284.10(M+H) + .
Step 2: preparation of 4- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) -3-methylaniline (3c)
The 1b was replaced with 3b and the remaining required starting materials, reagents and preparation were the same as in example 1, step 3, giving a tan solid. 1 H NMR(400MHz,Chloroform-d)δ7.56(d,J＝7.5Hz,1H),7.36(s,1H),7.09(d,J＝8.1Hz,1H),6.91(d,J＝2.5Hz,1H),6.67(d,J＝2.4Hz,1H),6.61(dd,J＝8.1,2.5Hz,1H),6.46(dd,J＝7.5,2.5Hz,1H),3.87(s,3H),2.05(s,3H). 13 C NMR(101MHz,Chloroform-d)δ157.48,147.22,146.52,139.65,132.47,131.70,124.22,123.79,117.98,116.71,112.71,107.09,94.90,55.52,19.83；MS(EI)m/z 254.12(M+H) + .
And step 3: n- (3-fluoro-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (3)
The 1c was replaced with 3c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid.
1 H NMR(500MHz,Chloroform-d)δ8.98(s,1H),7.75(d,J＝2.3Hz,1H),7.67(dd,J＝8.2,2.3Hz,1H),7.60(d,J＝7.5Hz,1H),7.47–7.41(m,3H),7.36(d,J＝8.3Hz,1H),6.92(d,J＝2.5Hz,1H),6.52(dd,J＝7.5,2.5Hz,1H),3.87(s,3H),2.19(s,3H). 13 C NMR(101MHz,Chloroform-d)δ157.80,154.23,147.88,146.94,139.55,137.13,132.16,131.96,125.41,124.11,122.79,122.39,118.27,116.99,112.75,107.60,95.01,55.59,20.09；MS(EI)m/z 393.11(M+H) + ；HRMS(ESI)calcd for C 20 H 16 N 4 O 5 (M+H) + :393.1193；found 393.1190.
Example 4
The structure and preparation process of N- (2-methoxy-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (4) are shown in the following
The preparation process comprises the following steps:
step 1: preparation of 7-methyl-2-phenylimidazo [1,2-a ] pyridine (4b)
The 1-bromo-4-nitrobenzene was replaced with 4-bromo-2-methoxy-1-nitrobenzene, and the remaining required starting materials, reagents and preparation were the same as in step 2 of example 1 to give a yellow solid. 1 H NMR(400MHz,Chloroform-d)δ8.20(d,J＝7.5Hz,1H),8.02(d,J＝8.8Hz,1H),7.67(s,1H),7.21–7.18(m,2H),6.95(d,J＝2.6Hz,1H),6.63(dd,J＝7.6,2.6Hz,1H),4.02(s,3H),3.90(s,3H)；MS(EI)m/z 300.09(M+H) + .
Step 2: preparation of 2-methoxy-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) aniline (4c)
The 1b was replaced with 4b and the remaining required starting materials, reagents and preparation were the same as in step 3 of example 1 to give a tan solid.
1 H NMR(400MHz,Chloroform-d)δ8.08(dd,J＝7.5,0.7Hz,1H),7.43(s,1H),6.94(dd,J＝7.9,1.8Hz,1H),6.91–6.87(m,2H),6.81(d,J＝7.9Hz,1H),6.50(dd,J＝7.5,2.5Hz,1H),3.89(s,3H),3.87(s,3H). 13 C NMR(101MHz,Chloroform-d)δ157.42,147.53,146.96,136.42,130.66,125.42,124.05,121.17,119.09,114.98,110.77,107.32,95.02,55.64,55.53；MS(EI)m/z 270.12(M+H) + .
And step 3: preparation of N- (2-methoxy-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (4)
The 1c was replaced with 4c and the remaining required starting materials, reagents and preparation were the same as in 4 of example 1 to give a tan solid.
1 H NMR(400MHz,Chloroform-d)δ8.92(s,1H),8.55(d,J＝8.3Hz,1H),8.17(d,J＝7.6Hz,1H),7.56(s,1H),7.43(d,J＝3.8Hz,1H),7.39(d,J＝3.8Hz,1H),7.19(dd,J＝8.3,1.8Hz,1H),7.07(d,J＝1.8Hz,1H),6.94(d,J＝2.5Hz,1H),6.57(dd,J＝7.6,2.5Hz,1H),4.04(s,3H),3.90(s,3H). 13 C NMR(126MHz,Chloroform-d)δ157.80,153.79,151.36,148.98,147.97,147.73,131.79,126.31,125.94,124.46,123.96,120.74,120.23,116.69,112.65,110.04,107.88,95.27,56.24,55.60；MS(EI)m/z 409.11(M+H) + ；HRMS(ESI)calcd for C 20 H 16 N 4 O 6 (M+H) + :409.1143；found 409.1148.
Example 5
The structure and preparation process of N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) -2-methylphenyl) -5-nitrofuran-2-carboxamide (5) are shown in the following
The preparation process comprises the following steps:
step 1: preparation of 7-methoxy-2-phenylimidazo [1,2-a ] pyridine (5b)
The 1-bromo-4-nitrobenzene was replaced with 4-bromo-2-methyl-1-nitrobenzene and the remaining required starting materials, reagents and preparation were the same as in step 2 of example 1 to give a tan solid.
1 H NMR(400MHz,Chloroform-d)δ8.21(d,J＝7.5Hz,1H),8.15(d,J＝9.1Hz,1H),7.67(s,1H),7.54–7.47(m,2H),6.96(d,J＝2.6Hz,1H),6.63(dd,J＝7.6,2.6Hz,1H),3.91(s,3H),2.71(s,3H). 13 C NMR(101MHz,Chloroform-d)δ158.24,148.90,147.51,135.32,134.57,134.00,130.70,126.09,124.77,123.89,122.89,108.47,95.63,55.68,21.07；MS(EI)m/z 248.10(M+H) + .
Step 2: preparation of 4- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) -2-methylaniline (5c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 5b, to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.06(d,J＝7.6Hz,1H),7.41(s,1H),7.21–7.10(m,2H),6.89(d,J＝2.6Hz,1H),6.77(d,J＝8.0Hz,1H),6.48(dd,J＝7.5,2.5Hz,1H),3.85(s,3H),2.22(s,3H). 13 C NMR(126MHz,Chloroform-d)δ157.39,146.90,144.82,130.49,130.47,127.07,125.31,124.08,122.85,119.06,115.24,107.22,94.96,55.50,17.43；MS(EI)m/z 254.12(M+H) + .
And step 3: n- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) -2-methylphenyl) -5-nitrofuran-2-carboxamide (5)
The 1c is changed to the 5c,the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,Chloroform-d)δ8.29(s,1H),8.20(s,1H),8.08(d,J＝8.2Hz,1H),7.46(m,5H),7.28(s,2H),6.96(s,1H),6.59(d,J＝7.6Hz,1H),3.91(s,3H),2.48(s,3H)；MS(EI)m/z 393.11(M+H) + ；HRMS(ESI)calcd for C 20 H 16 N 4 O 5 (M+H) + :393.1193；found 393.1195.
Example 6
The structure and preparation process of N- (3-methoxy-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (6) are shown in the following
The preparation process comprises the following steps:
step 1: preparation of 7-methoxy-3- (2-methoxy-4-nitrophenyl) imidazo [1,2-a ] pyridine (6b)
The 1-bromo-4-nitrobenzene was replaced with 1-bromo-2-methoxy-4-nitrobenzene, and the remaining required starting materials, reagents and preparation were the same as in step 2 of example 1 to give a tan solid.
1 H NMR(400MHz,Chloroform-d)δ7.97(dd,J＝8.3,2.2Hz,1H),7.89(d,J＝2.2Hz,1H),7.75(s,1H),7.58(d,J＝8.4Hz,1H),6.97(s,1H),6.57(d,J＝7.5Hz,1H),3.95(s,3H),3.90(s,3H)；MS(EI)m/z 300.09(M+H) + .
Step 2: preparation of 3-methoxy-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) aniline (6c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 6b, to give a tan solid. 1 H NMR(400MHz,Chloroform-d)δ7.64(dd,J＝7.5,0.7Hz,1H),7.41(s,1H),7.15(d,J＝7.9Hz,1H),6.91(d,J＝2.5Hz,1H),6.47(dd,J＝7.5,2.5Hz,1H),6.40–6.35(m,2H),3.88(s,3H),3.74(s,3H). 13 C NMR(126MHz,Chloroform-d)δ158.33,157.37,148.75,146.89,132.95,131.59,125.74,122.47,107.95,107.37,106.29,98.35,94.75,55.46,55.30；MS(EI)m/z 270.12(M+H) + .
And step 3: preparation of N- (3-methoxy-4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (6)
The 1c was replaced by 6c and the remaining required starting materials, reagents and preparation were the same as in 4 of example 1 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.82(s,1H),7.91–7.81(m,2H),7.71–7.65(m,2H),7.57(d,J＝8.2Hz,1H),7.48(s,1H),7.41(d,J＝8.1Hz,1H),7.02(s,1H),6.63(d,J＝7.0Hz,1H),3.87(s,3H),3.80(s,3H)；MS(EI)m/z 409.11(M+H) + ；HRMS(ESI)calcd for C 20 H 16 N 4 O 6 (M+H) + :409.1143；found 409.1141.
Example 7
The structure and preparation process of N- (5- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) pyridin-2-yl) -5-nitrofuran-2-carboxamide (7) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 5- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) pyridin-2-amine (7c)
The 1-bromo-4-nitrobenzene was replaced with 5-bromopyridin-2-amine and the remaining required starting materials, reagents and preparation were the same as in example 1, step 2, giving a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.23(s,1H),7.96(dd,J＝7.4,2.3Hz,1H),7.56(d,J＝8.5Hz,1H),7.46(s,1H),6.91(d,J＝2.5Hz,1H),6.63(dd,J＝8.5,2.2Hz,1H),6.53(dd,J＝7.4,2.7Hz,1H),4.67(s,2H),3.88(s,3H). 13 C NMR(126MHz,Chloroform-d)δ158.07,157.61,147.94,147.46,137.95,131.51,123.66,122.00,115.62,108.71,107.66,95.20,55.55；MS(EI)m/z 241.10(M+H) + .
Step 2: preparation of N- (5- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) pyridin-2-yl) -5-nitrofuran-2-carboxamide (7)
Changing 1c to 7c, the rest of the required raw materials, reagents and preparationsThe procedure is as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ11.48(s,1H),8.69(d,J＝2.4Hz,1H),8.52(d,J＝7.6Hz,1H),8.31(d,J＝8.6Hz,1H),8.19(dd,J＝8.7,2.4Hz,1H),7.93(d,J＝3.9Hz,1H),7.90–7.69(m,2H),7.12(d,J＝2.6Hz,1H),6.82(dd,J＝7.6,2.5Hz,1H),3.92(s,3H). 13 C NMR(101MHz,DMSO-d 6 )δ159.45,155.59,152.64,151.14,147.51,147.44,137.78,129.79,126.39,121.82,121.38,117.74,115.26,113.75,108.87,94.60,56.53；MS(EI)m/z 380.09(M+H) + ；HRMS(ESI)calcd for C 18 H 13 N 5 O 5 (M+H) + :380.0989；found 380.0989.
Example 8
The structure and preparation process of N- (3- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (8) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 3- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) aniline (8c)
The 1-bromo-4-nitrobenzene was replaced with 3-bromoaniline and the remaining required starting materials, reagents and preparation were the same as in example 1, step 2, to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.18(d,J＝7.5Hz,1H),7.50(d,J＝2.3Hz,1H),7.29–7.26(m,1H),6.90(t,J＝3.9Hz,2H),6.82(s,1H),6.71(d,J＝7.6Hz,1H),6.51(dd,J＝7.5,2.8Hz,1H),3.87(d,J＝2.3Hz,3H)；MS(EI)m/z 240.11(M+H) + .
Step 2: preparation of N- (3- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (8)
The 1c was replaced with 8c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ10.86(s,1H),8.59(d,J＝7.6Hz,1H),8.08(d,J＝3.9Hz,1H),8.03(s,1H),7.86(m,2H),7.67(d,J＝3.9Hz,1H),7.62(t,J＝8.0Hz,1H),7.47(d,J＝7.9Hz,1H),7.22(d,J＝2.6Hz,1H),7.04(dd,J＝7.6,2.6Hz,1H),3.98(s,3H). 13 C NMR(101MHz,DMSO-d 6 )δ161.43,155.32,152.31,148.14,144.56,139.11,130.51,127.61,127.05,125.44,124.87,121.62,120.50,117.37,113.99,110.25,93.39,57.02；MS(EI)m/z 379.10(M+H) + ；HRMS(ESI)calcd for C 19 H 14 N 4 O 5 (M+H) + :379.1037；found 379.1035.
Example 9
The structure and preparation process of N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) -3- (trifluoromethyl) phenyl) -5-nitrofuran-2-carboxamide (9) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 4- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) -3- (trifluoromethyl) aniline (9c)
The 1-bromo-4-nitrobenzene was replaced with 4-bromo-3- (trifluoromethyl) aniline and the remaining required starting materials, reagents and preparation were the same as in example 1, step 2, giving a tan solid. 1 H NMR(500MHz,Chloroform-d)δ7.52(d,J＝7.5Hz,1H),7.41(s,1H),7.18(dd,J＝8.3,2.6Hz,1H),7.09(d,J＝2.2Hz,1H),6.88(dt,J＝8.7,2.3Hz,2H),6.45(dt,J＝7.6,1.9Hz,1H),4.18(s,2H),3.86(d,J＝2.4Hz,3H).MS(EI)m/z 308.09(M+H) + .
Step 2: preparation of N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) -3- (trifluoromethyl) phenyl) -5-nitrofuran-2-carboxamide (9)
The 1c was replaced with 9c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,Acetone-d 6 )δ10.75(s,1H),8.48(d,J＝2.3Hz,1H),8.24(dd,J＝8.4,2.2Hz,1H),7.95(d,J＝7.5Hz,1H),7.69–7.61(m,3H),7.55(d,J＝3.9Hz,1H),7.06(d,J＝2.6Hz,1H),6.71(dd,J＝7.6,2.4Hz,1H),5.05(s,1H),3.91(s,3H)；MS(EI)m/z 447.08(M+H) + ；HRMS(ESI)calcd for C 20 H 13 N 4 O 5 F 3 (M+H) + :447.0911；found 447.0912.
Example 10
The structure and preparation process of N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) -2- (trifluoromethyl) phenyl) -5-nitrofuran-2-carboxamide (10) are shown below:
the preparation process comprises the following steps:
step 1: preparation of 4- (7-Methoxyimidazo [1,2-a ] pyridin-3-yl) -2- (trifluoromethyl) aniline (10c)
The 1-bromo-4-nitrobenzene was replaced with 4-iodo-2- (trifluoromethyl) aniline, and the remaining required starting materials, reagents and preparation were the same as in example 1, step 2, to give a tan solid. 1 H NMR(500MHz,Methanol-d4)δ7.94(d,J＝7.6Hz,1H),7.36(d,J＝2.2Hz,1H),7.28–7.23(m,2H),6.88(d,J＝8.5Hz,1H),6.78(d,J＝2.6Hz,1H),6.48(dd,J＝7.5,2.5Hz,1H),3.74(s,3H).MS(EI)m/z 308.09(M+H) + .
Step 2: preparation of N- (4- (7-methoxyimidazo [1,2-a ] pyridin-3-yl) -2- (trifluoromethyl) phenyl) -5-nitrofuran-2-carboxamide (10)
The 1c was replaced with 10c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ10.70(s,1H),8.53(d,J＝7.5Hz,1H),8.05–7.99(m,2H),7.84(d,J＝3.9Hz,1H),7.80(s,1H),7.69(d,J＝8.1Hz,1H),7.65(d,J＝3.9Hz,1H),7.08(d,J＝2.6Hz,1H),6.72(dd,J＝7.5,2.6Hz,1H),3.88(s,3H)；HRMS(ESI)calcd for C 20 H 13 N 4 O 5 F 3 (M+H) + :447.0911；found 447.0912.
Example 11
The structure and preparation process of N- (4- (7-bromoimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (11) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 7-bromoimidazo [1,2-a ] pyridine (11a)
The 4-methoxypyridin-2-amine was replaced with 4-bromopyridin-2-amine and the remaining required starting materials, reagents and preparation were the same as in example 1, step 1 to give a brown oil. 1 H NMR(400MHz,Chloroform-d)δ7.87(d,J＝7.2Hz,1H),7.67(d,J＝2.0Hz,1H),7.46(d,J＝12.8Hz,2H),6.73(dd,J＝7.2,2.0Hz,1H). 13 C NMR(126MHz,Chloroform-d)δ145.38,134.10,125.99,119.80,118.03,116.10,112.68；MS(EI)m/z 196.96(M+H) + .
Step 2: preparation of 7-bromo-3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (11b)
The procedure of example 1 was repeated except for changing 1a to 11a and 1-bromo-4-nitrobenzene to 1-iodo-4-nitrobenzene, and the remaining required starting materials, reagents and preparation were the same as in example 1, step 2, to give a yellow solid. 1 H NMR(400MHz,DMSO-d 6 )δ8.68(dd,J＝7.3,0.8Hz,1H),8.40–8.32(m,2H),8.11–8.03(m,2H),8.02–7.97(m,2H),7.20(dd,J＝7.3,2.1Hz,1H). 13 C NMR(126MHz,DMSO)δ147.34,146.67,136.07,135.54,128.18,126.26,124.93,124.38,120.15,119.18,117.27；MS(EI)m/z 317.98(M+H) + .
And 3, step 3: preparation of 4- (7-bromoimidazo [1,2-a ] pyridin-3-yl) aniline (11c)
The remaining required starting materials, reagents and preparation were the same as in example 1, step 3, replacing 1b with 11b, to give a white solid. 1 H NMR(500MHz,Chloroform-d)δ8.11(d,J＝7.3Hz,1H),7.83(s,1H),7.60(s,1H),7.29(m,2H),6.86(d,J＝7.3Hz,1H),6.80(d,J＝8.0Hz,2H). 13 C NMR(126MHz,Chloroform-d)δ147.00,132.01,129.68,129.59,123.87,120.18,118.20,117.54,116.15,115.49,113.89.MS(EI)m/z 288.00(M+H) + .
And 4, step 4: preparation of N- (4- (7-bromoimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (11)
Changing 1c to 11c, the rest being requiredThe starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ11.01(s,1H),8.54(d,J＝7.4Hz,1H),7.99(dd,J＝9.3,2.3Hz,3H),7.84(s,2H),7.79(s,1H),7.73–7.66(m,2H),7.10(dd,J＝7.3,2.0Hz,1H). 13 C NMR(126MHz,DMSO-d 6 )δ155.17,152.30,148.25,138.37,132.92,130.11,128.62,125.87,124.61,121.65,119.68,118.31,117.27,116.84,113.96.MS(EI)m/z 427.00(M+H) + ；HRMS(ESI)calcd for C 18 H 11 N 4 O 4 Br(M+H) + :427.0036；found 427.0033.
Example 12
The structure and preparation process of N- (4- (7-chloroimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (12) are shown as follows:
the preparation process comprises the following steps:
step 1: preparation of 7-chloroimidazo [1,2-a ] pyridine (12a)
The 4-methoxypyridin-2-amine was replaced with 4-chloropyridin-2-amine, and the remaining required starting materials, reagents and preparation were the same as in example 1, step 1 to give a brown oil. 1 H NMR(400MHz,CDCl 3 )δ8.07,8.06,7.64,7.64,7.64,7.58,7.57,6.80,6.80,6.79,6.78. 13 C NMR(126MHz,CDCl 3 )δ145.20,134.46,130.85,125.96,116.73,114.05,112.57；MS(EI)m/z 153.58(M+H) + .
Step 2: preparation of 7-chloro-3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (12b)
The remaining required starting materials, reagents and preparation were the same as in example 1, step 2, replacing 1a with 12a, to give a yellow solid. 1 H NMR(400MHz,DMSO-d 6 )δ8.74(dd,J＝7.3,0.8Hz,1H),8.39–8.34(m,2H),8.07(s,1H),8.03–7.98(m,2H),7.91(dd,J＝2.3,0.8Hz,1H),7.12(dd,J＝7.4,2.2Hz,1H). 13 C NMR(126MHz,DMSO-d 6 )δ147.01,146.68,136.24,135.55,131.26,128.19,126.38,124.94,124.38,116.90,114.96；MS(EI)m/z 274.03(M+H) + ..
And step 3: preparation of 4- (7-chloroimidazo [1,2-a ] pyridin-3-yl) aniline (12c)
The other required starting materials, reagents and preparation were the same as in example 1, step 3, replacing 1b with 12b, to give a white solid. 1 H NMR(400MHz,Chloroform-d)δ8.15(dd,J＝7.3,0.8Hz,1H),7.63(dd,J＝2.1,0.8Hz,1H),7.57(s,1H),7.35–7.27(m,2H),6.85–6.78(m,2H),6.75(dd,J＝7.4,2.1Hz,1H). 13 C NMR(126MHz,Chloroform-d)δ147.02,145.14,131.90,130.49,129.63,126.46,123.82,118.10,116.74,115.48,113.98；MS(EI)m/z 244.05(M+H) + .
And 4, step 4: preparation of N- (4- (7-chloroimidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (12)
The 1c was replaced with 12c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.83(s,1H),8.60(dd,J＝7.4,1.8Hz,1H),7.95(d,J＝8.3Hz,2H),7.86–7.83(m,2H),7.81(d,J＝1.5Hz,1H),7.70(dt,J＝12.1,3.1Hz,3H),7.02(dd,J＝7.4,2.1Hz,1H). 13 C NMR(126MHz,DMSO-d 6 )δ155.16,152.30,148.26,138.25,133.69,129.99,128.56,125.84,124.82,121.66,117.24,116.64,114.31,113.97；MS(EI)m/z 383.05(M+H) + ；HRMS(ESI)calcd for C 18 H 11 N 4 O 4 Cl(M+H) + :383.0542；found 383.0539.
Example 13
The structure and preparation process of N- (4- (7- (1-methylpiperidin-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (13) are as follows:
the preparation process comprises the following steps:
step 1: preparation of 7- (4-methylpiperazin-1-yl) -3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (13b)
11b (400mg, 1.26mmol) and 1-methylpiperazine (151mg, 1.51mmol) were added to a clean flask and dissolved in 10mL of N, N-dimethylformamide, and cesium carbonate (821mg, 2.52mmol), BINAP (235.11mg, 0.378mmol) and Pd (dba)3(115.29mg, 0.126mmol) were added to the flask under nitrogen at room temperature. After stirring at 110 ℃ for 12h, the reaction solution was diluted with 100ml of ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The resulting solution was evaporated and purified by silica gel column chromatography (dichloromethane: methanol 10:1, V/V) to give the target product as a red brown solid (230mg, 54%). 1 H NMR(500MHz,Chloroform-d)δ8.33(d,J＝8.5Hz,2H),8.24(d,J＝7.7Hz,1H),7.72–7.66(m,3H),6.87(d,J＝2.5Hz,1H),6.72(dd,J＝7.8,2.5Hz,1H),3.33(t,J＝5.1Hz,4H),2.60(t,J＝5.1Hz,4H),2.37(s,3H). 13 C NMR(126MHz,Chloroform-d)δ149.78,148.99,146.04,136.32,134.80,126.34,124.77,123.46,122.26,107.04,97.94,54.54,47.97,46.08；MS(EI)m/z 338.15(M+H) + .
Step 2: preparation of 4- (7- (4-methylpiperazin-1-yl) imidazo [1,2-a ] pyridin-3-yl) aniline (13c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 13b, to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.04(d,J＝7.7Hz,1H),7.40(s,1H),7.27(d,J＝8.3Hz,2H),6.86–6.75(m,3H),6.57(dd,J＝7.7,2.5Hz,1H),3.28–3.22(m,4H),2.58(t,J＝5.0Hz,4H). 13 C NMR(126MHz,CDCl 3 )δ148.09,147.42,146.35,130.65,129.26,124.55,123.49,119.39,115.45,106.38,98.15,54.68,48.51,46.10；MS(EI)m/z 308.17(M+H) + .
And step 3: preparation of N- (4- (7- (1-methylpiperidin-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (13)
The 1c was replaced by 13c and the remaining required starting materials, reagents and preparation were as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.90(s,1H),8.14(d,J＝7.6Hz,1H),7.85(d,J＝8.1Hz,2H),7.56–7.49(m,3H),7.43(q,J＝3.7Hz,2H),6.84(s,1H),6.65(dd,J＝7.7,2.3Hz,1H),3.29(t,J＝5.1Hz,4H),2.59(t,J＝5.0Hz,4H),2.36(s,3H). 13 C NMR(126MHz,Chloroform-d)δ154.18,151.25,148.64,147.85,136.06,131.22,128.29,126.53,123.59,123.50,121.25,116.97,112.72,106.74,97.59,54.60,48.22,46.09；MS(EI)m/z 447.17(M+H) + ；HRMS(ESI)calcd for C 19 H 13 N 4 O 5 F(M+H) + :447.1775；found 447.1773.
Example 14
The structure and preparation process of 5-nitro-N- (4- (7- (pyrrolidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) furan-2-carboxamide (14) are as follows:
the preparation method comprises the following specific steps:
step 1: preparation of 3- (4-Nitrophenyl) -7- (pyrrolidin-1-yl) imidazo [1,2-a ] pyridine (14b)
The 1-methylpiperazine was replaced by pyrrolidine, and the remaining required starting materials, reagents and preparation were the same as in step 1 of example 13 to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.33–8.27(m,2H),8.22(d,J＝7.4Hz,1H),7.68–7.63(m,3H),6.52–6.45(m,2H),3.43–3.36(m,4H),2.11–2.05(m,4H). 13 C NMR(126MHz,Chloroform-d)δ150.74,145.75,145.52,136.71,134.94,125.66,124.78,123.58,121.36,105.00,92.62,47.76,25.45；MS(EI)m/z 309.12(M+H) + .
Step 2: preparation of 4- (7- (pyrrolidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) aniline (14c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 14b, to give a tan solid. 1 H NMR(400MHz,Chloroform-d)δ8.02(d,J＝7.6Hz,1H),7.31(d,J＝12.5Hz,2H),7.28(s,1H),6.86–6.72(m,2H),6.50(d,J＝2.4Hz,1H),6.40(dd,J＝7.6,2.4Hz,1H),3.41–3.34(m,4H),2.08–2.03(m,4H). 13 C NMR(126MHz,Chloroform-d)δ147.34,146.42,145.61,129.36,127.57,123.73,123.62,118.99,115.48,104.49,91.27,47.83,25.42；MS(EI)m/z 279.15(M+H) + .
And step 3: preparation of 5-nitro-N- (4- (7- (pyrrolidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) furan-2-carboxamide (14)
The reaction mixture 1c was replaced with 14c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.89(s,1H),8.45(d,J＝7.7Hz,1H),8.03–7.96(m,2H),7.93(s,1H),7.85(d,J＝3.8Hz,1H),7.76–7.66(m,3H),6.91(dd,J＝7.8,2.4Hz,1H),6.46(d,J＝2.4Hz,1H),3.46(s,4H),2.07–1.99(m,4H). 13 C NMR(126MHz,DMSO-d 6 )δ155.29,152.34,149.49,148.11,142.96,139.41,129.66,127.10,124.51,122.09,121.59,118.37,117.42,113.98,107.79,85.90,48.38,25.38；MS(EI)m/z 418.1510(M+H) + ；HRMS(ESI)calcd for C 22 H 19 N 5 O 4 (M+H) + :418.1510；found 418.1510.
Example 15
The structure and preparation process of N- (4- (7- (1, 1-dioxy thiomorpholinyl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (15) are as follows:
the preparation process comprises the following steps:
step 1: preparation of 4- (3- (4-nitrophenyl) imidazo [1,2-a ] pyridin-7-yl) thiomorpholine 1, 1-dioxide (15b)
The 1-methylpiperazine was replaced by 1, 1-dithiomorpholine, and the remaining required starting materials, reagents and preparation were the same as in example 13, step 1, giving a tan solid. 1 H NMR(400MHz,Chloroform-d)δ8.29(d,J＝7.7Hz,1H),8.06–7.96(m,2H),7.83–7.80(m,1H),7.37(dd,J＝6.8,4.7Hz,2H),6.75(dd,J＝7.9,2.5Hz,2H),4.14(q,J＝7.1Hz,4H),3.74(d,J＝7.1Hz,4H)；MS(EI)m/z 373.08(M+H) + .
Step 2: preparation of 4- (3- (4-aminophenyl) imidazo [1,2-a ] pyridin-7-yl) thiomorpholine 1, 1-dioxide (15c)
Changing 1b to 15b, and the restThe required raw materials, reagents and preparation method are the same as the step 3 in the example 1, and a brown yellow solid is obtained. 1 H NMR(400MHz,Chloroform-d)δ8.11(d,J＝7.7Hz,1H),7.44(s,1H),7.29(d,J＝2.0Hz,1H),7.27(s,1H),6.91(d,J＝2.4Hz,1H),6.83–6.77(m,2H),6.53(dd,J＝7.7,2.6Hz,1H),3.93–3.87(m,4H),3.17–3.12(m,4H). 13 C NMR(126MHz,Chloroform-d)δ146.71,146.64,144.25,131.15,129.38,125.11,124.52,118.80,115.48,105.57,99.51,50.31,47.18；MS(EI)m/z 343.63(M+H) + .
And step 3: preparation of N- (4- (7- (1, 1-dioxothiomorpholinyl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (15)
The 1c was replaced with 15c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.88(s,1H),8.42(d,J＝7.6Hz,1H),7.94(d,J＝8.4Hz,2H),7.84(d,J＝3.9Hz,1H),7.76(d,J＝3.9Hz,1H),7.67–7.60(m,3H),6.99(s,2H),3.91(t,J＝5.0Hz,4H),3.21–3.17(m,4H). 13 C NMR(101MHz,DMSO-d 6 )δ155.10,152.29,148.34,147.49,145.66,137.68,131.34,130.11,127.91,125.28,123.75,121.71,117.22,113.95,106.65,97.45,50.30,46.68；MS(EI)m/z 482.11(M+H) + ；HRMS(ESI)calcd for C 22 H 19 N 5 O 6 S(M+H) + :482.1129；found 482.1130.
Example 16
The structure and preparation process of N- (4- (7- (4-cyclopropylpiperazin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (16) are as follows:
the preparation method comprises the following specific steps:
step 1: preparation of 7- (4-Cyclopropylpiperazin-1-yl) -3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (16b)
Example 13 was repeated except for 1-cyclopropylpiperazine instead of 1-methylpiperazine and the remaining required starting materials, reagents and preparation methodsStep 1 of (1), a tan solid was obtained. 1 H NMR(400MHz,DMSO-d 6 )δ8.55(d,J＝7.7Hz,1H),8.30(d,J＝8.7Hz,2H),7.91(d,J＝8.6Hz,2H),7.85(s,1H),7.03–6.96(m,1H),6.86(s,1H),3.26(d,J＝5.2Hz,4H),2.68(t,J＝5.0Hz,4H),2.13–1.90(m,1H),0.45(m,2H),0.36(m,2H)；MS(EI)m/z 364.16(M+H) + .
Step 2: preparation of 4- (7- (4-cyclopropylpiperazin-1-yl) imidazo [1,2-a ] pyridin-3-yl) aniline (16c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 16b, to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ8.34(d,J＝7.8Hz,1H),7.80(s,1H),7.28(d,J＝8.1Hz,3H),6.97(s,1H),6.75(d,J＝8.2Hz,2H),2.04–1.95(m,1H),0.85(m,2H),0.62(s,2H)；MS(EI)m/z 334.19(M+H) + .
And step 3: preparation of N- (4- (7- (4-cyclopropylpiperazin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (16)
The 1c was replaced with 16c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.84(s,1H),8.39(d,J＝7.7Hz,1H),7.93(d,J＝8.3Hz,2H),7.84(d,J＝3.9Hz,1H),7.71(d,J＝3.9Hz,1H),7.68–7.63(m,3H),6.99(dd,J＝7.8,2.4Hz,1H),6.78(d,J＝2.4Hz,1H),3.28(t,J＝5.2Hz,4H),2.69(t,J＝5.0Hz,4H),1.71–1.66(m,1H),0.49–0.44(m,2H),0.39–0.35(m,2H). 13 C NMR(126MHz,DMSO-d 6 )δ155.12,152.29,149.61,148.28,146.63,137.95,130.11,128.22,125.29,124.82,123.85,121.64,117.22,113.97,107.25,94.60,52.76,47.67,38.45,6.10.；MS(EI)m/z 473.19(M+H) + ；HRMS(ESI)calcd for C 25 H 24 N 6 O 4 (M+H) + :473.1932；found 473.1931.
Example 17
The structure and preparation process of N- (4- (7- (4-hydroxy-4-methylpiperidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (17) are shown below
The preparation process comprises the following steps:
step 1: preparation of 4-methyl-1- (3- (4-nitrophenyl) imidazo [1,2-a ] pyridin-7-yl) piperidin-4-ol (17b)
The 1-methylpiperazine was replaced by 4-methylpiperidin-4-ol and the remaining required starting materials, reagents and preparation were the same as in example 13, step 1 to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.38–8.28(m,2H),8.23(d,J＝7.7Hz,1H),7.68-7.70(m,3H),6.90(d,J＝2.5Hz,1H),6.74(dd,J＝7.7,2.5Hz,1H),3.55–3.47(m,2H),3.38–3.29(m,2H),1.77–1.73(m,4H),1.33(s,3H). 13 C NMR(126MHz,Chloroform-d)δ149.99,148.98,146.00,136.36,134.60,126.28,124.79,123.47,122.18,107.38,97.63,67.65,44.77,37.80,30.20.MS(EI)m/z 353.15(M+H) + .
Step 2: preparation of 4-methyl-1- (3- (4-nitrophenyl) imidazo [1,2-a ] pyridin-7-yl) piperidin-4-ol (17c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 17b, to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ8.13(d,J＝7.7Hz,1H),7.28(s,1H),7.21(d,J＝8.2Hz,2H),6.81(dd,J＝7.8,2.5Hz,1H),6.70(m,3H),5.32(brs,2H),3.42–3.39(m,2H),3.22–3.16(m,2H),1.60–1.54(m,4H),1.16(s,3H). 13 C NMR(126MHz,DMSO-d 6 )δ148.87,147.92,147.26,129.93,128.87,124.59,124.17,116.70,114.72,106.82,96.46,66.34,44.92,37.94,30.13.MS(EI)m/z 323.17(M+H) + .
And 3, step 3: the structure for the preparation of N- (4- (7- (4-hydroxy-4-methylpiperidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (17) is shown below
The 1c was replaced with 17c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.88(s,1H),8.33(d,J＝7.7Hz,1H),7.92(d,J＝8.2Hz,2H),7.83(d,J＝3.8Hz,1H),7.75(d,J＝3.9Hz,1H),7.62(d,J＝8.2Hz,2H),7.51(s,1H),6.87(d,J＝7.8Hz,1H),6.74(s,1H),3.44(d,J＝17.0Hz,2H),3.24–3.18(m,2H),1.57(t,J＝5.4Hz,4H),1.16(s,3H). 13 C NMR(126MHz,DMSO-d 6 )δ155.05,152.27,148.37,148.30,137.31,131.97,130.10,127.59,126.00,124.52,123.30,121.68,117.14,113.97,107.03,96.34,66.35,44.75,37.89,30.14.MS(EI)m/z 462.17(M+H) + ；HRMS(ESI)calcd for C 24 H 23 N 5 O 5 (M+H) + :462.1772；found 462.1770.
Example 18
The structure and preparation process of N- (4- (7- (4-methyl-1, 4-diaza-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (18) are as follows:
the preparation process comprises the following steps:
step 1: preparation of 7- (4-methyl-1, 4-diaza-1-yl) -3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (18b)
The 1-methylpiperazine was replaced by N-methylpiperazine, and the remaining required starting materials, reagents and preparation were the same as in example 13, step 1, giving a tan solid.
Step 2: 4- (7- (4-methyl-1, 4-diaza-1-yl) imidazo [1,2-a ] pyridin-3-yl) aniline (18c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 18b, to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ7.91(d,J＝7.7Hz,1H),7.24(s,1H),7.17(d,J＝8.2Hz,2H),6.69(d,J＝8.0Hz,2H),6.52(d,J＝2.6Hz,1H),6.36(dd,J＝7.8,2.6Hz,1H),4.20–4.04(m,2H),3.53–3.49(m,2H),3.44(t,J＝6.3Hz,2H),2.64–2.60(m,2H),2.49–2.45(m,2H),2.29(s,3H),1.95–1.90(m,2H). 13 C NMR(126MHz,Chloroform-d)δ146.97,145.41,145.32,128.27,128.10,122.70,118.25,114.41,102.49,91.33,56.66,55.91,47.88,47.33,45.66,26.42.MS(EI)m/z 322.19(M+H) + .
And step 3: preparation of N- (4- (7- (4-methyl-1, 4-diaza-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (18)
The 1c was replaced with 18c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ11.25(s,1H),8.33(d,J＝7.7Hz,1H),8.11(d,J＝3.9Hz,1H),8.02(d,J＝8.3Hz,2H),7.83(d,J＝3.9Hz,1H),7.59(d,J＝8.3Hz,2H),7.47(s,1H),6.75(d,J＝7.8Hz,1H),6.53(s,1H),3.53–3.48(m,4H),2.70(t,J＝4.9Hz,2H),2.55(d,J＝5.0Hz,2H),2.31(s,3H),1.96–1.91(m,2H)；MS(EI)m/z 461.19(M+H) + ；HRMS(ESI)calcd for C 19 H 13 N 4 O 5 F(M+H) + :461.1932；found 461.1933.
Example 19
The structure and preparation process of N- (4- (7- (4-hydroxy-4-methylpiperidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (19) are as follows:
the preparation process comprises the following steps:
step 1: preparation of 3- (4-Nitrobenzene) -7- (4- (oxyethan-3-yl) piperazin-1-yl) imidazo [1,2-a ] pyridine (19b)
The 1-methylpiperazine was replaced by 1- (3-oxetanyl) piperazine and the remaining required starting materials, reagents and preparation were the same as in example 13, step 1, giving a tan solid. 1 H NMR(400MHz,Chloroform-d)δ8.35(d,J＝8.3Hz,2H),8.25(d,J＝7.6Hz,1H),7.69(m,3H),7.27(s,2H),6.91(s,1H),6.74(d,J＝7.7Hz,1H),4.69(dt,J＝23.6,6.3Hz,4H),3.57(m,1H),3.37(t,J＝4.9Hz,4H),2.53(t,J＝4.9Hz,4H).MS(EI)m/z 380.16(M+H) + .
Step 2: preparation of 4- (7- (4- (oxyethan-3-yl) piperazin-1-yl) imidazo [1,2-a ] pyridin-3-yl) aniline (19c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 19b, to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ8.31(d,J＝7.9Hz,1H),7.78(s,1H),7.27(d,J＝8.4Hz,3H),6.94(d,J＝2.5Hz,1H),6.78–6.71(m,2H),4.59(q,J＝6.3Hz,4H),3.57(s,4H),3.39(s,1H),2.57(s,4H).MS(EI)m/z 350.19(M+H) + .
And step 3: preparation of N- (4- (7- (4-hydroxy-4-methylpiperidin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (19)
The 1c was replaced with 19c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ10.86(s,1H),8.42(d,J＝7.8Hz,1H),7.95(d,J＝8.4Hz,2H),7.84(d,J＝3.9Hz,1H),7.76(s,1H),7.71(d,J＝4.0Hz,1H),7.67(d,J＝8.6Hz,2H),7.08(dd,J＝7.9,2.4Hz,1H),6.83(s,1H),4.59(t,J＝6.5Hz,2H),4.49(t,J＝6.0Hz,2H),3.41(m,5H),2.43(t,J＝5.1Hz,4H). 13 C NMR(101MHz,DMSO)δ155.22,152.32,151.01,148.19,138.88,130.33,130.16,128.97,126.23,124.47,123.28,122.14,117.35,113.97,107.67,94.04,74.73,58.77,49.02,46.94.MS(EI)m/z 489.18(M+H) + ；HRMS(ESI)calcd for C 25 H 24 N 6 O 5 (M+H) + :489.1881；found 489.1885.
Example 20
The structure and preparation process of tert-butyl 4- (3- (4- (5-nitrofuran-2-carboxamide) phenyl) imidazo [1,2-a ] pyridin-7-yl) piperazine-1-carboxylate (20) are as follows:
the preparation process comprises the following steps:
step 1: preparation of tert-butyl 4- (3- (4-nitrophenyl) imidazo [1,2-a ] pyridin-7-yl) piperazine-1-carboxylate (20b)
The 1-methylpiperazine was replaced by piperazine-1-carboxylic acid tert-butyl ester, and the remaining required raw materials, reagents and preparation method were the same as in example 13, step 1, yielding a tan solid. 1 H NMR(400MHz,Chloroform-d)δ8.36(d,J＝8.8Hz,2H),8.27(d,J＝7.7Hz,1H),7.75–7.67(m,3H),6.92(d,J＝2.4Hz,1H),6.77–6.70(m,1H),3.64(t,J＝5.2Hz,4H),3.29(t,J＝5.2Hz,4H),1.52(s,9H). 13 C NMR(126MHz,Chloroform-d)δ154.57,149.44,148.95,146.22,136.14,134.57,126.55,124.80,123.62,122.44,107.39,98.43,80.33,48.14,43.22,27.67.MS(EI)m/z 424.19(M+H) + .
Step 2: 4- (3- (4-aminophenyl) imidazo [1,2-a ] pyridin-7-yl) piperazine-1-carboxylic acid tert-butyl ester (20c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 20b, to give a tan solid. 1 H NMR(400MHz,Methanol-d 4 )δ8.01(d,J＝7.5Hz,1H),7.25(s,1H),7.16(d,J＝8.1Hz,2H),6.79(d,J＝8.1Hz,2H),6.69–6.62(m,2H),3.49(t,J＝5.3Hz,4H),3.14(d,J＝5.3Hz,4H),1.47(s,9H). 13 C NMR(101MHz,Methanol-d 4 )δ154.82,149.01,148.35,146.57,128.83,127.24,125.36,124.00,116.79,115.11,106.59,95.14,80.09,43.49,42.41,27.36.MS(EI)m/z 394.21(M+H) + .
And step 3: preparation of tert-butyl 4- (3- (4- (5-nitrofuran-2-carboxamide) phenyl) imidazo [1,2-a ] pyridin-7-yl) piperazine-1-carboxylate (20)
The 1c was replaced with 20c and the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ10.90(s,1H),8.50(d,J＝7.7Hz,1H),7.99(d,J＝8.1Hz,3H),7.85(d,J＝3.9Hz,1H),7.71(dd,J＝6.3,2.3Hz,3H),7.22(d,J＝7.9Hz,1H),6.86(s,1H),3.57–3.49(m,8H),1.44(s,9H). 13 C NMR(126MHz,DMSO-d 6 )δ155.26,154.29,152.32,152.03,148.12,143.05,139.37,129.56,127.16,122.16,121.57,117.39,113.96,108.00,89.23,79.76,54.06,46.43,28.50.MS(EI)m/z 533.20(M+H) + ；HRMS(ESI)calcd for C 27 H 29 N 6 O 6 (M+H) + :533.2068；found 533.2064.
Example 21
The structure and preparation process of 5-nitro-N- (4- (7- (piperazine-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) furan-2-carboxamide (21) are as follows:
the preparation process comprises the following steps:
step 1: preparation of 5-nitro-N- (4- (7- (piperazin-1-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) furan-2-carboxamide (21)
Compound 20(50mg, 0.09mmol) was dissolved in 5mL of dichloromethane, and 1mL of trifluoroacetic acid was added dropwise to the above reaction solution at room temperature. After stirring at room temperature for 1 hour, the solution was diluted with 50mL of dichloromethane. A saturated sodium bicarbonate solution was added while cooling on ice and the pH was adjusted to 7. The organic layer was washed with brine and dried over anhydrous sodium sulfate. The resulting solution was evaporated and purified by silica gel column chromatography (dichloromethane/methanol 50:3, V/V) to give the desired product (30mg, 77%) as a yellow-brown solid. 1 H NMR(500MHz,DMSO-d 6 )δ10.89(s,1H),8.50(d,J＝7.8Hz,1H),7.99(t,J＝4.5Hz,3H),7.85(d,J＝3.9Hz,1H),7.70(dd,J＝10.0,6.1Hz,3H),7.23(d,J＝8.2Hz,1H),6.86(s,1H),3.54(t,J＝10.1Hz,8H).MS(EI)m/z 433.15(M+H) + ；HRMS(ESI)calcd for C 20 H 21 N 6 O 4 (M+H) + :433.1540；found 433.1544.
Example 22
The structure and preparation process of tert-butyl 4- (3- (4- (5-nitrofuran-2-carboxamide) phenyl) imidazo [1,2-a ] pyridin-7-yl) -3, 6-dihydropyridine-1 (2H) -carboxylate (2) are as follows:
the preparation process comprises the following steps:
step 1: preparation of 4- (3- (4-Nitrophenyl) imidazo [1,2-a ] pyridin-7-yl) -3, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (22b)
Compound 11b (736mg, 2mmol) and N-Boc-1,2,5, 6-tetrahydropyridine-4-boronic acid pinacol ester (679.8mg, 2.2mmol) were dissolved in 10mL of a 1, 4-dioxane/water (3:1, V/V) solution, and potassium carbonate (424mg, 4mmol) and [1,1' -bis (diphenylphosphino) ferrocene were dissolved at room temperature]Palladium dichloride dichloromethaneThe complex (81.6mg, 0.1mmol) was added to the above reaction solution. After stirring at 110 ℃ for 12h, the reaction was diluted with 100ml of ethyl acetate, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. The resulting solution was evaporated and purified by silica gel column chromatography (dichloromethane/methanol 50:1, V/V) to give 545mg of a brown solid, yield, 65%. 1 H NMR(500MHz,Chloroform-d)δ8.12(d,J＝7.4Hz,1H),7.30(d,J＝9.0Hz,2H),7.26(d,J＝8.0Hz,2H),6.90(d,J＝7.4Hz,1H),6.81(d,J＝7.9Hz,2H),6.21(s,1H),4.11(s,2H),3.56(d,J＝6.0Hz,2H),2.44(s,2H),1.49(s,9H).MS(EI)m/z 421.18(M+H) + .
Step 2: 4- (3- (4-aminophenyl) imidazo [1,2-a ] pyridin-7-yl) -3, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (22c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 22b, to give a tan solid. 1 H NMR(500MHz,Chloroform-d)δ8.16(d,J＝7.4Hz,1H),7.55(d,J＝9.0Hz,2H),7.31(d,J＝8.0Hz,2H),6.90(d,J＝7.4Hz,1H),6.81(d,J＝7.9Hz,2H),6.21(s,1H),4.13(s,2H),3.66(d,J＝6.0Hz,2H),2.56(s,2H),1.50(s,9H).MS(EI)m/z 391.21(M+H) + .
And step 3: preparation of tert-butyl 4- (3- (4- (5-nitrofuran-2-carboxamide) phenyl) imidazo [1,2-a ] pyridin-7-yl) -3, 6-dihydropyridine-1 (2H) -carboxylate (2)
The 1c was replaced with 22c and the remaining required starting materials, reagents and preparation were the same as in 4 of example 1 to give a tan solid. 1 H NMR(400MHz,Chloroform-d)δ8.48(s,1H),8.25(d,J＝7.3Hz,1H),7.79(d,J＝8.2Hz,2H),7.67(s,1H),7.63(s,1H),7.52(s,1H),7.47(d,J＝8.1Hz,2H),7.24(d,J＝3.2Hz,1H),7.14(d,J＝7.1Hz,1H),6.54(s,1H),6.31(s,1H),4.11(s,2H),3.62(s,2H),2.50(s,2H),1.48(s,9H).MS(EI)m/z 530.19(M+H) + ；HRMS(ESI)calcd for C 28 H 28 N 5 O 6 (M+H) + :530.1968；found 530.1964.
Example 23
The structure of 5-nitro-N- (4- (7- (1,2,3, 6-tetrahydropyridin-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) furan-2-carboxamide (23) is shown below:
the preparation process comprises the following steps:
step 1: preparation of 5-nitro-N- (4- (7- (1,2,3, 6-tetrahydropyridin-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) furan-2-carboxamide (23)
The 20 was changed to 22 and the remaining required starting materials, reagents and preparation were the same as in example 21, step 1, giving a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ11.22(s,1H),9.62(s,1H),8.56(d,J＝7.4Hz,1H),8.06(m,3H),7.85(d,J＝3.9Hz,1H),7.80(s,1H),7.70(d,J＝8.5Hz,2H),7.67(s,1H),7.21(d,J＝7.0Hz,1H),6.48(s,1H),3.83–3.72(m,2H),3.05–2.59(m,2H),2.51(s,2H). 13 C NMR(126MHz,DMSO-d 6 )δ155.13,152.29,148.30,146.47,137.93,136.52,133.39,132.60,128.20,125.43,125.37,125.21,123.99,121.68,117.21,113.97,111.85,110.48,45.01,42.59,25.99.MS(EI)m/z 430.14(M+H) + ；HRMS(ESI)calcd for C 23 H 20 N 5 O 4 (M+H) + :430.1437；found 430.1435.
Example 24
The structure and preparation process of N- (4- (7- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (24) are as follows:
the preparation process comprises the following steps:
step 1: preparation of N- (4- (7- (1-acetyl-1, 2,3, 6-tetrahydropyridin-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (24)
The remaining required starting materials, reagents and preparation were the same as in example 1, step 4, replacing 1c with 30, to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ8.53(d,J＝7.4Hz,1H),8.16(m,3H),7.85(d,J＝3.9Hz,1H),7.81(s,1H),7.70(d,J＝8.5Hz,2H),7.64(s,1H),7.18(d,J＝7.0Hz,1H),6.48(s,1H),3.83–3.72(m,2H),3.05–2.59(m,2H),2.51(s,2H),2.31(s,3H).MS(EI)m/z 472.15(M+H) + ；HRMS(ESI)calcd for C 25 H 22 N 5 O 5 (M+H) + :472.1544；found 472.1539.
Example 25
The structure and preparation process of N- (4- (7- (1-methyl-1H-pyrazol-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (25) are as follows:
the preparation process comprises
Step 1: preparation of 7- (1-methyl-1H-pyrazol-4-yl) -3- (4-nitrophenyl) imidazo [1,2-a ] pyridine (25b)
The N-Boc-1,2,5, 6-tetrahydropyridine-4-boronic acid pinacol ester was replaced with 1-methyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxabenzaldehyde-2-yl) -1H-pyrazole, and the remaining required starting materials, reagents and preparation were the same as in step 1 of example 22 to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ8.46(s,1H),8.07(s,1H),8.19(d,J＝8.2Hz,2H),7.99(m,2H),7.71(s,1H),7.68(d,J＝8.5Hz,2H),7.21(d,J＝7.2Hz,1H),3.91(s,3H)；MS(EI)m/z 320.10(M+H) + ；
Step 2: preparation of 4- (7- (1-methyl-1H-pyrazolyl-4-yl) imidazo [1,2-a ] pyridin-3-yl) aniline (25c)
The remaining required starting materials, reagents and procedures were the same as in example 1, step 3, replacing 1b with 25b, to give a tan solid. 1 H NMR(500MHz,DMSO-d 6 )δ8.36(s,1H),8.05(s,1H),8.01(d,J＝8.2Hz,2H),7.86(m,2H),7.71(s,1H),7.69(d,J＝8.5Hz,2H),7.20(d,J＝7.2Hz,1H),3.90(s,3H)；MS(EI)m/z 290.13(M+H) + ；
And step 3: preparation of N- (4- (7- (1-methyl-1H-pyrazol-4-yl) imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (25)
Changing 1c to 25c, and remaining required raw materials and reagentsAnd the preparation method is the same as the step 4 in the example 1, and a brown yellow solid is obtained. 1 H NMR(500MHz,DMSO-d 6 )δ11.12(s,1H),8.56(d,J＝7.2Hz,1H),8.36(s,1H),8.08(s,1H),8.03(d,J＝8.2Hz,2H),7.97(d,J＝2.9Hz,1H),7.89–7.84(m,2H),7.73(s,1H),7.70(d,J＝8.5Hz,2H),7.22(d,J＝7.2Hz,1H),3.90(s,3H)；MS(EI)m/z 429.12(M+H) + ；HRMS(ESI)calcd for C 22 H 17 N 6 O 4 (M+H) + :429.1230；found 429.1230.
Example 26
The structure and preparation process of N- (4- (imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (26) are shown as follows:
the preparation process comprises the following steps:
step 1: the structure for the preparation of N- (4- (imidazo [1,2-a ] pyridin-3-yl) phenyl) -5-nitrofuran-2-carboxamide (26c) is shown below
By replacing 1c with 4- (imidazo [1,2-a ]]Pyridin-3-yl) aniline, the remaining required starting materials, reagents and preparation were the same as in step 4 of example 1 to give a tan solid. 1 H NMR(400MHz,DMSO-d 6 )δ10.82(s,1H),8.59(d,J＝6.9Hz,1H),7.95(d,J＝8.6Hz,2H),7.83(d,J＝3.9Hz,1H),7.78(s,1H),7.71(s,1H),7.70–7.68(m,2H),7.66(d,J＝9.1Hz,1H),7.31(q,1H),6.98(t,J＝6.5Hz,1H).MS(EI)m/z 349.08(M+H) + ；HRMS(ESI)calcd for C 18 H 13 N 4 O 4 (M+H) + :349.0854；found 349.0856.
Biological Activity test section
It was found that compounds 1-19 and compound 23 were able to inhibit the proliferation of a variety of cancer cells, including breast, lung and gastric cancer cells.
The compound 23 small molecule compound can effectively inhibit the tyrosine phosphorylation level of STAT3, and inhibit the transcriptional activity of STAT3 and the expression of downstream target genes Bcl-xl, C-myc and Mcl-1 thereof.
The compound 23 small molecular compound can obviously inhibit the growth of gastric cancer in a mouse subcutaneous cell transplantation tumor model, reduces the tumor body weight and has no obvious toxicity to mouse organs.
Application example 1: compounds 1-19 and 23 inhibit breast, lung and gastric cancer cell proliferation
(1) Cell culture
MDA-MB-231 and MCF-7 cells are cultured in DMEM medium containing 10% fetal calf serum and 1% double-resistant penicillin and streptomycin, lung cancer cells A549 and PC9, gastric cancer cells AGS and MGC803, melanoma cells MP41 and 92-1, leukemia cells MOLM-13 and MV4-11 and prostate cancer cells C42 are cultured in RPIM-1640 medium containing 10% fetal calf serum and 1% double-resistant penicillin and streptomycin, and the cells are placed at 37 ℃ and 5% CO 2 In a cell culture incubator.
(2) CCK8 method for determining cell viability
1) Taking each cell in logarithmic phase, digesting the cell, counting, uniformly inoculating the cell in a 96-well plate at the density of 100 mu L per well of (1-3) x 10000/well, and placing the cell in a constant temperature incubator until the cell adheres to the wall;
2) after adherence, adding corresponding culture medium into a control group, only adding the culture medium into a blank group without cells, and adding various compounds with different concentrations into an administration group;
3) after the drug treatment is carried out for 72h, the cell state is observed under a microscope, CCK8(10 mu L/hole) is added under the condition of keeping out of the sun, and the mixture is evenly mixed and placed in an incubator at 37 ℃ in the absence of the light for incubation for 0.5-4 h;
4) after bubbles are removed, reading the light absorption value (OD) at the position of 450nm of an enzyme label, and repeating the experiment for three times;
5) cell viability (%) ═ a (compound) -a (blank) ]/[ a (control) -a (blank) ] × 100%;
6) finally, calculation was performed using GraphPad Prism8 software to obtain IC 50.
(3) Results of the experiment
The experimental results are shown in table 1, and the compounds 1 to 19 and the compound 23 have good activity on 4 cancer cells and show good inhibition effect; FIG. 1 shows the growth inhibition curves and IC50 of compound 23 on various cancer cells, including lung cancer cells A549 and PC9, gastric cancer cells AGS and MGC803, melanoma cells MP41 and 92-1, leukemia cells MOLM-13 and MV4-11, and prostate cancer cell C42.
TABLE 1 inhibitory Activity of Compounds 1-19 and 23 against 4 cancer cells
Application example 2: compound 23 is capable of inhibiting phosphorylation of STAT3 and dimer formation
The present application example uses compound 23 as an example, and measures the effect of inhibiting phosphorylation of STAT3 and formation of a dimer. The specific process is as follows:
(1) western blotting (Western Blot)
After the cells are treated by the compound 23 with different concentrations for 24 hours, the culture medium is sucked off, the cell lysate RIPA containing protease inhibitor and phosphatase inhibitor is added to lyse the cells, then the lysate is centrifuged for 15min at 15000rpm and 4 ℃, the supernatant is taken out for quantification, and 5 × loading buffer is added to the lysate for boiling denaturation. Then separating protein samples by using polyacrylamide gel SDS-PAGE electrophoresis, transferring the protein samples to a nitrocellulose membrane (PVDF membrane), after blocking for 1h by 5% BSA, respectively incubating overnight at 4 ℃ by using primary antibodies of pY705-STAT3, STAT3, C-myc, Bcl-xl, Mcl1 and beta-Actin, respectively, then incubating for 1h at room temperature by using a rabbit secondary antibody and a mouse secondary antibody which are provided with fluorescent markers, and finally detecting the expression level of the protein by using a Tanon developing instrument.
(2) Results of the experiment
Experimental results as shown in fig. 2, fig. (2A) shows that compound 23 is capable of time-dependently inhibiting phosphorylation of STAT3(pY 705); figure (2B) shows that compound 23 is able to inhibit STAT3(pY705) phosphorylation concentration-dependently.
Application example 3: compound 23 can inhibit STAT3 transcriptional activity and target gene expression
In this application, compound 23 was used as an example, and the transcriptional activity of STAT3 and the expression effect of the target gene were measured. The specific process is as follows:
(1) western blotting method is as above
(2) Dual luciferase reporter gene assay
1) Taking 293T cells in a logarithmic growth phase, digesting the cells, counting the cells, and inoculating the cells into a 96-well plate at the density of 1-2 x 10000 cells/100 mu L;
2) culturing the cells in an incubator at 37 ℃ overnight, and performing a cell transfection experiment when the cell fusion degree is about 80%;
3) a, B two mixed solutions were prepared in 1.5mL centrifuge tubes (hereinafter, the amount of each well)
A: 0.25. mu.L Lipo 2000+ 5. mu.L OPTI-MEMI Medium
B: 50ng pGL3-STAT3-promoter plasmid +50ng STAT3C plasmid +40ng TKRL (Renilla luciferin reporter plasmid) + 5. mu.L OPTI-MEM Medium
Mixing solution A and solution B, and standing at room temperature for 5 min;
4) gently mixing the A, B solutions, and standing at room temperature for 15 min;
5) adding the mixed solution into each hole of a 96-hole plate along the 45-degree angle of the wall;
6) putting 96-well plate cells into a cell culture box, incubating for 24h, and adding compounds with different concentrations for 23 treatment for 24 h;
7) dissolving firefly luciferase detection reagent and Renilla luciferase detection buffer solution to room temperature, and placing Renilla luciferase detection substrate (100X) on ice bath for later use;
8) taking a proper amount of renilla luciferase detection buffer solution according to the dosage of 100 mu Lyongli per hole, and adding a renilla luciferase detection substrate (100X) according to the ratio of 1:100 to prepare renilla luciferase detection working solution;
9) taking out the 96-well plate, discarding the culture medium, adding 50 μ L of reporter gene cell lysate into each well, shaking and mixing for 5 min;
10) adding 50 mu L of firefly luciferase detection reagent into each hole, and shaking and uniformly mixing for 5 min;
11) detecting to obtain RLU 1;
12) after the step of measuring the firefly luciferase is completed, 100 mu L of renilla luciferase detection working solution is added into each hole, and the solution is shaken and uniformly mixed for 5 min;
13) detecting to obtain RLU 2;
14) the ratio RLU1/RLU2 was obtained.
(3) Results of the experiment
The experimental results are shown in fig. 3, and fig. 3A shows that compound 23 can significantly inhibit the transcriptional activity of STAT 3; FIG. 3B shows that Compound 23 is capable of downregulating phosphorylation of STAT3(pY705) and its protein expression of the target genes C-myc, Bcl-xl and Mcl-1.
Application example 4: the compound 23 can obviously inhibit the growth of tumors in stomach cancer bodies
In this application, compound 23 was used as an example to measure inhibition of tumor growth in stomach cancer. The specific process is as follows:
(1) subcutaneous tumor-bearing experiment in nude mice
1) And (3) taking the gastric cancer cells MGC803 in the logarithmic growth phase, digesting, counting, mixing the precooled PBS and Matrigel according to the ratio of 1:1, re-suspending the cells to obtain 2 x 106 cells/100 mu L of cell suspension, and placing the cell suspension on ice. Injecting 100 mu L of cell suspension into subcutaneous parts on two sides of the abdomen and the back of a 4-5-week-old immunodeficient nude mouse respectively;
2) when the subcutaneous tumor volume is about 100mm3, the subcutaneous tumors are divided into three groups randomly. Drug solvents (20% castor oil in PBS), 3mg/kg of Compound 23 in the low dose group, and 10mg/kg of Compound 23 in the high dose group were administered to the control group. The administration mode is intraperitoneal injection, the administration volume is 100 mu L/unit, the administration is carried out once a day, and the administration is carried out continuously for four weeks. The mouse body weight and tumor size were measured every two days and mouse body weight gain and tumor growth curves were plotted.
3) Four weeks after dosing, mice were sacrificed for dissection, subcutaneous tumors were dissected, weighed and photographed.
(2) Results of the experiment
The results of the experiment are shown in fig. 4, where (4A) is the final volume size of representative tumors in each group after four weeks of administration; fig. 4B is a graph of tumor volume change during dosing, demonstrating that compound 23 is able to significantly inhibit the growth of gastric cancer tumors dose-dependently; FIG. 4C shows that Compound 23 is capable of dose-dependently reducing gastric cancer tumor weight, with a tumor inhibition of about 37.2% in the 3mg/kg group and about 78.86% in the 10mg/kg group; FIG. 4D is a graph showing the body weight change of nude mice during administration, indicating that Compound 23 has no effect on the body weight of nude mice.
Application example 5: compound 23 inhibits expression of pY705-STAT3 protein and target gene in vivo
In this application, the compound 23 was used as an example, and the inhibition of the expression of pY705-STAT3 protein and the target gene in vivo was measured. The specific process is as follows:
(1) hematoxylin-eosin staining method (HE)
The heart, liver, spleen, lung, kidney and subcutaneous tumors of each group of mice were stripped, washed with PBS, fixed with paraformaldehyde, dehydrated, paraffin-embedded, sliced, stained with hematoxylin and eosin, dehydrated and mounted, observed under a microscope and photographed, and the effect of compound 23 on visceral tissues and stomach cancer tumors was examined.
(2) Western blotting experiment (Western Blot)
Tumor tissue (about 20mg) from mice cryopreserved with liquid nitrogen was removed, thawed on ice, and 200. mu.L of RIPA lysate (containing protease inhibitor and phosphatase inhibitor) was added to the frozen tumor tissue, and the tissue pieces were cut into small pieces on ice using tissue scissors. And (3) fully cracking the tissue blocks by using ice ultrasound (Apml 45%), stopping ultrasound when no tissue blocks are visible to the naked eyes, centrifuging at the temperature of 4 ℃ and the speed of 15000rpm for 15-20min, taking supernatant into a new centrifuge tube, quantifying BCA, and carrying out subsequent experiments by using the Western blot as described above.
(3) Results of the experiment
Experimental results as shown in fig. 5, fig. (5A) shows that compound 23 is also able to inhibit phosphorylation of STAT3(pY705) and protein expression of its target gene in mouse in vivo experiments; FIG. 5B is a HE diagram showing the tumor exfoliated in groups of nude mice, showing significant increase in tumor tissue space and cell arrangement disorder in the group to which compound 23 was administered; FIG. 5C is a HE chart of heart, liver, spleen, lung and kidney peeled from each group of nude mice, showing that the cell arrangement distribution, shape and the like of each tissue were not significantly different among the groups.
It should be finally noted that the above examples are only intended to illustrate the technical solutions of the present invention, and not to limit the scope of the present invention, and that other variations and modifications based on the above description and thought may be made by those skilled in the art, and that all embodiments need not be exhaustive. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (7)
1. The nitrofuran compound is characterized by having a structure shown as a formula (I):
wherein:
x is C or N;
R 1 、R 2 、R 3 、R 4 independently selected from hydrogen, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 1-6 Alkylamino, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 3-8 Cycloalkoxy, substituted or unsubstituted C 3-8 Cycloalkylamino, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted 3-to 8-membered heterocyclyl with 1 to 4 heteroatoms selected from N and O, -COR a 、-CO 2 R a 、-CONR a R b 、-NR a C(O)R b 、-NR a SO 2 R b 、-SR a 、-SOR a 、-SO 2 R a 、-SO 2 NR a R b 、-OC(O)R a or-OC (O) NR a R b ；
R a And R b Independently of one another is hydrogen, C 1-6 Alkyl or C 3-6 A cycloalkyl group;
R 5 is hydrogen, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, substituted or unsubstituted C 1-8 Alkyl, substituted or unsubstituted C 1-8 Alkoxy, substituted or unsubstituted C 1-8 Alkylamino, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or substituted C 3-8 Cycloalkoxy, substituted or unsubstituted C 1-8 Alkylamino, substituted or unsubstituted aryl or heteroaryl;
R 6 、R 7 、R 8 、R 9 independently selected from hydrogen, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, C 1-3 Alkoxy radical, C 1-3 An alkylamino group;
R 1 、R 2 、R 3 、R 4 is substituted at least at 1 position with the following substituents: halogen, cyano, amino, nitro, hydroxy, trifluoromethyl, C 1-3 Alkyl radical, C 1-3 Alkoxy radical, C 1-3 Alkylamino, -COR a 、-CO 2 R a 、-CONR a R b 、-NR a C(O)R b 、-NR a SO 2 R b 、-SR a 、-SOR a 、-SO 2 R a 、-SO 2 NR a R b 、-OC(O)R a 、-OC(O)NR a R b ；
R 5 Is substituted at least at 1 position with the following substituents: halogen, cyano, amino, nitro, hydroxy, trifluoromethyl, methylthio, C 1-3 Alkyl radical, C 1-3 Alkoxy or C 1-3 An alkylamino group;
R 1 、R 2 、R 3 、R 4 、R 5 wherein said aryl or heteroaryl is independently selected from the group consisting of: furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, benzofuryl, benzothienyl, benzoxazolyl, benzothiazolyl, phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, quinolinyl, or naphthyl.
2. According to claimThe nitrofurans of claim 1, wherein R is 1 、R 2 、R 3 、R 4 Independently selected from H, halogen, cyano, nitro, amino, hydroxy, trifluoromethyl, C 1-6 Alkyl radical, C 1-6 Heteroalkylalkoxy, C 1-6 Alkylamino radical, C 3-8 Cycloalkyl radical, C 3-8 Cycloalkoxy, C 3-8 Cycloalkylamino, aryl, heteroaryl, 3-to 8-membered heterocyclyl containing 1-2 heteroatoms selected from N and O.
3. A method for preparing a nitrofuran compound as claimed in any one of claims 1 to 2, which comprises the following steps:
s1: dissolving the formula (1) and the formula (2) in a solvent, and generating an intermediate (3) after reaction;
s2: inserting a side chain into the intermediate (3) to generate an intermediate (4);
s3: reducing the intermediate (4) to generate an intermediate (5), and then carrying out condensation reaction to obtain the nitrofuran compound;
4. use of the nitrofurans compound or the pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 in preparation of a medicament for inhibiting STAT3 protein activity.
5. The use according to claim 4, wherein the nitrofurans or pharmaceutically acceptable salts thereof are used for preparing a medicament for treating abnormal cell proliferation, morphological change and hyperkinesia of STAT3 high expression, or treating angiogenesis or cancer metastasis;
the application of the nitrofurans or the pharmaceutically acceptable salt thereof in preparing the medicines for inhibiting the tyrosine phosphorylation level of STAT 3.
6. Use of the nitrofurans compound of claim 1 to 2 or a pharmaceutically acceptable salt thereof in preparing a medicament for inhibiting tumor growth and proliferation.
7. A pharmaceutical composition comprising the nitrofurans of claims 1 to 2 or a pharmaceutically acceptable salt thereof, and an EGFR inhibitor.
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