7-sulfur-containing substituent-containing chromone-3-carboxaldehyde compounds or derivatives thereof and preparation method and application thereof

Abstract

The application discloses a 7-sulfur-substituted chromone-3-formaldehyde compound shown in a formula I or a derivative thereof shown in a formula II and a preparation method and application thereof, wherein R is selected from a methylthio group, a thiazole-2-yl group, a pyrimidine-2-yl group, a pyridine-2-yl group, an imidazole-2-yl group, a (1-methyl-pyridine-4) group-4-thiazole-2-yl group, a benzothiazole-2-yl group, a quinoline-2-yl group, a benzothiophene-2-yl group, a (4-carboxylpropyl)-5-thiophene-2-yl group, and n represents the number of S; when R is selected from a thiophene-2-yl group, n is selected from 0, 1 or 2; when R is selected from groups other than the thiophene-2-yl group, n is selected from 0. The compound and the derivative thereof have a better inhibiting effect on NDM-1, can enhance the antibacterial effect in combination with imipenem and meropenem, and in addition, the synthesis route is simple and easy to operate.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, and particularly relates to a 7-sulfur-containing substituent-chromone-3-formaldehyde compound or a derivative thereof, and a preparation method and application thereof. BACKGROUND

[0002] β-lactam antibiotics (penicillins, cephalosporins, carbapenems, monobactams) are a class of important antibacterial drugs for treating diseases caused by bacterial infections, and are widely used in clinic. However, with the extensive use of antibiotics, bacteria gradually develop drug resistance, and the most important resistance mechanism is to produce β-lactamase that can hydrolyze β-lactam antibiotics. New Delhi metallo-β-lactamase (NDM-1) is a metallo-β-lactamase reported in 2009, which can hydrolyze all β-lactam antibiotics except monobactams, and confer multiple drug resistance to bacteria (ANTIMICROB. AGENTS. CH., 2009, 53(12), 5046-5054). The plasmid with the NDM-1 gene can spread among different bacteria, causing rapid spread of drug resistance, and posing a great threat to global public health (CURR. OPIN. MICROBIOL., 2010, 13, 558-564).

[0003] An effective strategy against β-lactamase-induced drug resistance is to develop safe and effective β-lactamase inhibitors to be used in combination with β-lactam antibiotics to inhibit bacterial drug resistance. At present, various types of NDM-1 inhibitors have been reported. Captopril, an angiotensin-converting enzyme inhibitor, is clinically used to treat hypertension and heart failure, and is found to have inhibitory effect on NDM-1 (Protein Cell, 2011, 2(5), 384-394). The natural product aspergillomarasmine A produces an inhibitory effect by stripping the zinc ions from the active site of NDM-1, and can overcome the drug resistance of NDM-1-producing bacteria when used in combination with meropenem, restoring the therapeutic effect of meropenem (Nature, 2014, 510, 5003-506). Quinoline-2-carboxylic acid compounds can tightly bind to the zinc-containing active center of NDM-1, thereby producing strong inhibitory effect and making the multi-drug resistant bacteria expressing NDM-1 sensitive to meropenem again (J. Med. Chem., 2023, 66, 11761-11791). Boronic acid compounds can form a tetrahedral structure similar to the enzyme-substrate complex with the active center of NDM-1, and have inhibitory effect on both serine-β-lactamase and metallo-β-lactamase, and have the potential to become a broad-spectrum β-lactamase inhibitor (J. Med. Chem., 2020, 63, 7491-7507). Ebselen is a glutathione peroxidase mimic with anti-inflammatory, antioxidant and anticancer activity, and is found to form a covalent S-Se bond with the cysteine residues in the active site of NDM-1 to produce an inhibitory effect (Chem. Commun. 2015, 51, 9543-9546). Although there are many studies on NDM-1 inhibitors at present, there is no clinically available NDM-1 inhibitor, so it is still an important task to develop new safe and effective NDM-1 inhibitors.

[0004] Chromone compounds are a class of unique natural products widely existing in animals and plants, and are concerned due to their potential anticancer, anti-inflammatory, antioxidant, antibacterial and other biological activities. Among them, chromone-3-carboxaldehyde is found to have NDM-1 inhibitory activity (Bioorg. Med. Chem., 2016, 24:2947-2953). Therefore, how to design and synthesize specific compounds with NDM-1 inhibitory activity based on chromone-3-carboxaldehyde as the skeleton is an important idea to solve the drug resistance caused by β-lactamase.

[0005] The present application is a research carried out under the support of the National Natural Science Foundation (21702239). SUMMARY

[0006] The present application aims to provide a 7-sulfur-containing substituent-chromone-3-formaldehyde compound or its derivative, and a preparation method and application thereof. Based on the crystal structure of NDM-1 enzyme and the skeleton characteristics of chromone-3-formaldehyde, a 7-substituted sulfur / chalcone-3-formaldehyde compound and its derivative are designed and synthesized, which has a good inhibitory effect on New Delhi metallo-beta-lactamase 1 (NDM-1), and provides a new scheme for developing a clinically applicable NDM-1 inhibitor.

[0007] The present application is implemented by the following technical solutions:

[0008] The 7-sulfur-containing substituent-chromone-3-formaldehyde compound shown in formula I or the derivative of the 7-sulfur-containing substituent-chromone-3-formaldehyde compound shown in formula II:

[0009]

[0010] wherein R is selected from methylthio, thiazole-2-yl, pyrimidine-2-yl, pyridine-2-yl, imidazole-2-yl, (1-methyl-pyridine-4)yl-4-thiazole-2-yl, benzothiazole-2-yl, quinoline-2-yl, benzothiophene-2-yl, (4-carboxylpropyl)-5-thiophene-2-yl, and n represents the number of S connected; when R is selected from thien-2-yl, n is selected from 0, 1 or 2; when R is selected from groups other than thien-2-yl, n is selected from 0.

[0011] When n is selected from 0, the structure of the 7-sulfur-containing substituent-chromone-3-formaldehyde compound shown in formula I is as follows:

[0012] The preparation method comprises the following steps:

[0013] The 4-sulfur-substituted-2-hydroxyacetophenone intermediate (compound A) is used as a raw material, and is reacted in a POCl3 and N,N-dimethylformamide system at 20-40°C for 6-18 hours to obtain the target 7-substituted sulfur-chromone-3-formaldehyde compound (compound B); the molar ratio of compound A and POCl3 is 1:1.5-5, and the reaction formula is as follows:

[0014]

[0015] Preferably, the concentration of compound A is 0.1-0.5 mol / L.

[0016] The preparation method of the 4-sulfur-substituted-2-hydroxyacetophenone intermediate (compound A) is as follows:

[0017] ​The compound A is obtained by using 4-mercapto-2-hydroxyacetophenone and aromatic iodide as raw materials, in the presence of potassium hydroxide and cuprous oxide, using dimethyl sulfoxide and water as mixed solvents, and reacting at 90-130 °C for 6-18 hours; the molar ratio of 4-mercapto-2-hydroxyacetophenone, aromatic iodide, potassium hydroxide and cuprous oxide is 1:(1-2):(1.5-5):(0.02-0.2); the volume ratio of dimethyl sulfoxide and water in the mixed solvent is 4:1-0.2, and the reaction formula is as follows:

[0018]

[0019] or,

[0020] The compound A is obtained by using 4-mercapto-2-hydroxyacetophenone and aromatic iodide as raw materials, in the presence of potassium hydroxide and cuprous oxide, using dimethyl sulfoxide and water as mixed solvents, and reacting at 90-130 °C for 6-18 hours; the molar ratio of 4-mercapto-2-hydroxyacetophenone, aromatic iodide, potassium hydroxide and cuprous oxide is 1:(1-2):(1.5-5):(0.02-0.2), the volume ratio of dimethyl sulfoxide and water in the mixed solvent is 4:1-0.2; the reaction formula is as follows:

[0021]

[0022] Preferably, the concentration of 4-mercapto-2-hydroxyacetophenone is 0.1-1 mol / L; the concentration of 4-iodo-2-hydroxyacetophenone is 0.2-2 mol / L.

[0023] In particular, when the R substituent in the 4-thio-substituted-2-hydroxyacetophenone intermediate (compound A) is thiazol-2-yl, the preparation method of the compound A-1 is as follows: 4-fluoro-2-hydroxyacetophenone and thiophene-2-thiol are used as raw materials, potassium carbonate is used as a base in N,N-dimethylformamide solvent, and the reaction is carried out at 90-130 °C for 1-6 hours to obtain 4-(thiophene-2-thio)yl-2-hydroxyacetophenone; the molar ratio of the 4-fluoro-2-hydroxyacetophenone, thiophene-2-thiol and potassium carbonate is 1:(1-2):(1-2); the reaction formula is as follows:

[0024]

[0025] Preferably, the concentration of 4-fluoro-2-hydroxyacetophenone is 0.1-1 mol / L.

[0026] In particular, when the R substituent in the 4-thio-substituted-2-hydroxyacetophenone intermediate (compound A) is (1-methyl-pyridin-4)yl-4-thiazol-2-yl, the preparation method of the compound is as follows:

[0027]

[0028] The preparation method of the 7-sulfur-containing substituent group-chromone-3-carboxaldehyde compound shown in formula I is as follows: when n is selected from 1 or 2, the 7-(thiophene-2-sulfenyl)-chromone-3-carboxaldehyde obtained by the above preparation method is used as a raw material, dichloromethane is used as a solvent, and the reaction is carried out at 20-30°C for 1-6 hours in the presence of 3-chloroperbenzoic acid; the molar ratio of the compound B and the 3-chloroperbenzoic acid is 1:1-2. The reaction formula is as follows:

[0029]

[0030] Preferably, the concentration of the 7-(thiophene-2-sulfenyl)-chromone-3-carboxaldehyde is 0.1-0.5 mol / L.

[0031] The preparation method of the derivative of the 7-sulfur-containing substituent group-chromone-3-carboxaldehyde compound shown in formula II is as follows: the 7-sulfur-containing substituent group-chromone-3-carboxaldehyde compound shown in formula I is used as a raw material, and the reaction is carried out in a tetrahydrofuran solvent at 20-40°C for 0.5-4 hours with a sodium bisulfite aqueous solution to obtain the derivative II; the molar ratio of the 7-sulfur-containing substituent group-chromone-3-carboxaldehyde compound and the sodium bisulfite is 1:1-2.5.

[0032] Preferably, the concentration of the 7-(thiophene-2-sulfenyl / sulfone / sulfoxide)-chromone-3-carboxaldehyde is 0.02-0.2 mol / L.

[0033] The application also protects the use of the 7-sulfur-containing substituent group-chromone-3-carboxaldehyde compound shown in formula I or the derivative thereof shown in formula II as an inhibitor of New Delhi metallo-beta-lactamase NDM-1 in the preparation of a medicine for inhibiting the activity of NDM-1, so as to alleviate the drug resistance of bacteria to beta-lactam antibiotics caused by NDM-1.

[0034] The beneficial effects of the application are as follows:

[0035] 1. The synthesis route of the application is simple and easy to operate, the raw materials are economical and easy to obtain, and the target compound with a chromone-3-carboxaldehyde parent structure can be quickly obtained; further, the corresponding aldehyde compound is protected by sodium bisulfite, thereby increasing the water solubility and stability.

[0036] 2. The 7-sulfur-containing substituent group-chromone-3-carboxaldehyde compound or the derivative thereof synthesized in the application has a good inhibitory effect on NDM-1 which causes bacteria to have multiple drug resistance, and the half-inhibitory concentration (IC 50 ) of the compound 23 is as low as 0.037 μM, which is about 1000 times higher than the inhibitory activity (IC 50 = 33.2 μM) of the positive control captopril.

[0037] 3、The 7-sulfur-containing substituent-chromone-3-carboxaldehyde compound or its derivative synthesized by the application has a synergistic effect when combined with the antibiotic imipenem (IMP) or combined with meropenem (MEM), and the minimum inhibitory concentration (MIC) of the antibiotic to the NDM-1-producing E. coli is reduced. Among them, the effect of compound 23 is the most obvious, which reduces the MIC of imipenem from 32 mg / L to 2 mg / L, and reduces the MIC of meropenem from 32 mg / L to 0.5 mg / L, and is expected to treat drug-resistant bacterial infections caused by NDM-1, and has good drug application prospect. DETAILED DESCRIPTION

[0038] Those skilled in the art will appreciate that the technology disclosed in the following examples represents the technology discovered by the inventors to function well in the practice of the application. However, many changes can be made in the specific embodiments disclosed, and yet obtain the same or similar results, without departing from the spirit and scope of the application.

[0039] The following is a further description of the application, rather than a limitation of the application.

[0040] I. Instruments and reagents

[0041] The nuclear magnetic resonance (NMR) of the application was determined by AVANCE 600 instrument produced by Bruker Company in Germany, and the solvent peak was used as an internal standard; the mass spectrum of the application was determined by Thermo UltiMate3000 ISQ EC (ESI source) produced by Thermo Fisher Company in the United States; chemical reagents were purchased from Shanghai Bide Pharmaceutical Technology Co., Ltd., J&K Company, Alfar-Aser Company, Aldrin Chemical Reagent Company, Shanghai Macklin Company, etc.; silica gel for column chromatography was purchased from Qingdao Marine Chemical Plant.

[0042] Example 1: Synthesis of intermediate A-1

[0043]

[0044] Method A: 4-fluoro-2-hydroxyacetophenone (0.154 g, 1 mmol) and 2-thiophene thiol (0.116 g, 1 mmol) were dissolved in N,N-dimethylformamide (10 mL), and potassium carbonate (0.138 g, 1 mmol) was added, and the reaction was carried out at 90°C for 6 hours. After the reaction was completed, the white solid was purified by silica gel column chromatography; 0.195 g, yield 78%.

[0045] Method B: 4-Fluoro-2-hydroxyacetophenone (0.154 g, 1 mmol) and 2- thiophene thiol (0.232 g, 2 mmol) were dissolved in N,N-dimethylformamide (1 mL), potassium carbonate (0.276 g, 2 mmol) was added, and the reaction was stirred at 130 °C for 1 h. After the reaction was completed, the product was purified by silica gel column chromatography to give a white solid; 0.223 g, 89% yield.

[0046] 1 H NMR (600 MHz, chloroform-d) δ 12.42 (s, 1H), 7.60 - 7.55 (m, 2H), 7.34 (d, J = 2.4 Hz, 1H), 7.14 (dd, J = 5.4, 3.6 Hz, 1H), 6.63 (dd, J = 8.5, 1.9 Hz, 1H), 6.59 (d, J = 1.9 Hz, 1H), 2.56 (s, 3H). MS (EI): m / z (%) = 250 (M + , 85), 235 ([M-CH3] + , 100).

[0047] Example 2: Synthesis of intermediate A-2

[0048]

[0049] 4-Mercapto-2-hydroxyacetophenone (0.168 g, 1 mmol) and 2-iodobenzothiophene (0.26 g, 1 mmol) were dissolved in dimethyl sulfoxide (1 mL), potassium hydroxide (0.168 g, 3 mmol), cuprous oxide (0.0072 g, 0.05 mmol), and water (0.2 mL) were added, and the reaction was stirred at 130 °C for 6 h. The reaction was cooled to room temperature, the pH was adjusted to weakly acidic, extracted, concentrated, and purified by silica gel column chromatography to give a white solid; 0.186 g, 62% yield.

[0050] 1 H NMR (600 MHz, chloroform-d) δ 12.39 (s, 1H), 7.92 (d, J = 8.1 Hz, 1H), 7.87 (s, 1H), 7.75 (d, J = 7.2 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.41 (dq, J = 14.9, 7.0 Hz, 2H), 6.59 (dd, J = 8.5, 1.8 Hz, 1H), 6.54 (d, J = 1.9 Hz, 1H), 2.54 (s, 3H). MS (ESI + ): m / z = 301 ([M+H] + ).

[0051] Example 3: Synthesis of intermediate A-3

[0052]

[0053] The synthesis method refers to Example 2 to obtain white solid, 0.131 g, yield 39%.

[0054] 1 H NMR (600 MHz, chloroform-d) δ 12.44 (s, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.17 (d, J = 3.5 Hz, 1H), 6.85 (d, J = 3.5 Hz, 1H), 6.64 (dd, J = 8.4, 1.9 Hz, 1H), 6.60 (d, J = 1.9 Hz, 1H), 2.92 (t, J = 7.5 Hz, 2H), 2.57 (s, 3H), 2.46 (t, J = 7.3 Hz, 2H), 2.09 - 2.01 (m, 2H). MS (ESI - ): m / z = 335 ([M-H] - ).

[0055] Example 4: Synthesis of intermediate A-4

[0056]

[0057] 4-Iodo-2-hydroxyacetophenone (0.262 g, 1 mmol), 2-mercaptothiazole (0.117 g, 1 mmol) were dissolved in dimethyl sulfoxide (1 mL), water (0.2 mL), potassium hydroxide (0.168 g, 3 mmol) and cuprous oxide (0.007 g, 0.05 mmol) were added, the reaction was heated to 120 °C for 6 hours. The reaction was cooled to room temperature, water was added, the pH was adjusted to weakly acidic, extracted, concentrated, purified by silica gel column chromatography to obtain colorless oil; 0.213 g, yield 85%.

[0058] 1 H NMR (600 MHz, chloroform-d) δ 12.4 (s, 1H), 7.9 (d, J = 3.4 Hz, 1H), 7.7 (d, J = 8.4 Hz, 1H), 7.5 (d, J = 3.4 Hz, 1H), 6.9 (d, J = 1.9 Hz, 1H), 6.9 (dd, J = 8.4, 1.9 Hz, 1H), 2.6 (s, 3H). MS (ESI + ): m / z = 252 ([M+H] + ).

[0059] Example 5: Synthesis of intermediate A-5

[0060]

[0061] Synthesis method reference example 4, yellowish liquid, 0.153 g, yield 62%.

[0062] 1 H NMR (600 MHz, chloroform-d) δ 12.31 (s, 1H), 8.54 (d, J = 4.8 Hz, 2H), 7.76 (d, J = 8.3 Hz, 1H), 7.32 (d, J = 1.7 Hz, 1H), 7.16 (dd, J = 8.3, 1.8 Hz, 1H), 7.05 (t, J = 4.8 Hz, 1H), 2.65 (s, 3H). MS (ESI + ): m / z = 247 ([M+H] + ).

[0063] Example 6: Synthesis of intermediate A-6

[0064]

[0065] Synthesis method reference example 4, yellowish liquid, 0.153 g, yield 62%.

[0066] 1 H NMR (600 MHz, chloroform-d) δ 12.35 (s, 1H), 8.54 (d, J = 4.9 Hz, 1H), 7.68 (d, J = 8.3 Hz, 1H), 7.62 (td, J = 7.7, 1.9 Hz, 1H), 7.31 (d, J = 8.0 Hz, 1H), 7.19 - 7.15 (m, 1H), 7.04 (d, J = 1.7 Hz, 1H), 6.96 (dd, J = 8.4, 1.7 Hz, 1H), 2.62 (s, 3H). MS (ESI + ): m / z = 246 ([M+H] + ).

[0067] Example 7: Synthesis of intermediate A-7

[0068]

[0069] Synthesis method reference example 4, white solid, 0.181 g, yield 77%.

[0070] 1H NMR (600 MHz, DMSO-d6) δ 13.00 (s, 1H), 12.10 (s, 1H), 7.80 (d, J = 8.5 Hz, 1H), 7.45 (s, 1H), 7.21 (s, 1H), 6.60 (dd, J = 8.4, 1.9 Hz, 1H), 6.46 (d, J = 1.9 Hz, 1H), 2.57 (s, 3H). MS (ESI + ): m / z = 235 ([M+H] + ).

[0071] Example 8: Synthesis of intermediate A-8

[0072]

[0073] Synthesis method refers to Example 4, yellow solid, 0.279g, yield 85%.

[0074] 1 H NMR (600 MHz, chloroform-d) δ 12.38 (s, 1H), 8.68 (d, J = 6.3 Hz, 2H), 7.81 - 7.77 (m, 3H), 7.71 (d, J = 8.4 Hz, 1H), 7.07 (d, J = 2.0 Hz, 1H), 7.00 (dd, J = 8.4, 2.0 Hz, 1H), 2.63 (s, 3H). MS (ESI + ): m / z = 329 ([M+H] + ).

[0075] Example 9: Synthesis of intermediate A-9

[0076]

[0077] The reaction equation is as follows:

[0078]

[0079] Method A: A-8 (0.164g, 0.5mmol) was dissolved in tetrahydrofuran (5mL), iodomethane (0.71g, 5mmol) was added, and the reaction was carried out at 20°C for 48 hours in the dark. After filtration, washing and drying, the product was obtained. Yellow solid, 0.179g, yield 76%.

[0080] Method B: A-8 (0.164g, 0.5mmol) was dissolved in tetrahydrofuran (1mL), iodomethane (0.142g, 10mmol) was added, and the reaction was carried out at 30°C for 24 hours in the dark. After filtration, washing and drying, the product was obtained. Yellow solid, 0.15g, yield 64%.

[0081] 1H NMR (600 MHz, DMSO-d6) δ 11.95 (s, 1H), 9.05 (s, 1H), 9.01 (d, J = 6.8 Hz, 2H), 8.57 (d, J = 6.9 Hz, 2H), 7.94 (d, J = 8.3 Hz, 1H), 7.22 - 7.14 (m, 2H), 4.34 (s, 3H), 2.65 (s, 3H). MS (ESI + ): m / z = 343 ([M-I - ] + ).

[0082] Example 10: Synthesis of intermediate A-10

[0083]

[0084] Synthesis method according to example 4, white solid, 0.23 g, yield 76%.

[0085] 1 H NMR (600 MHz, chloroform-d) δ 12.36 (s, 1H), 7.98 (d, J = 8.4 Hz, 1H), 7.79 - 7.72 (m, 2H), 7.50 - 7.45 (m, 1H), 7.40 - 7.34 (m, 1H), 7.24 (d, J = 1.7 Hz, 1H), 7.12 (dd, J = 8.3, 1.8 Hz, 1H), 2.65 (s, 3H). MS (ESI + ): m / z = 302 ([M+H] + ).

[0086] Example 11: Synthesis of intermediate A-11

[0087]

[0088] Synthesis method according to example 4, white solid, 0.155 g, yield 52%.

[0089] 1 H NMR (600 MHz, chloroform-d) δ 12.36 (s, 1H), 8.03 (d, J = 8.5 Hz, 2H), 7.77 (d, J = 8.8 Hz, 1H), 7.73 - 7.69 (m, 2H), 7.53 (t, J = 7.5 Hz, 1H), 7.28 (d, J = 8.6 Hz, 1H), 7.19 (d, J = 1.8 Hz, 1H), 7.07 (dd, J = 8.4, 1.8 Hz, 1H), 2.64 (s, 3H). MS (ESI + ): m / z = 296 ([M+H] + ).

[0090] Example 12: Synthesis of compound 1

[0091]

[0092] The product of Example 1, A-1 (0.125 g, 0.5 mmol) was dissolved in N,N- dimethylformamide (4 mL) and cooled in an ice water bath. Phosphorous oxychloride (0.23 g, 1.5 mmol) was added slowly and the reaction was allowed to warm to 25 °C for 18 h. The reaction was poured into ice water and the pH was adjusted to neutral. The mixture was extracted and concentrated. The product was purified by silica gel column chromatography to give a light yellow solid; 0.112 g, 90% yield.

[0093] 1 H NMR (600 MHz, chloroform-d) δ 10.35 (s, 1H), 8.43 (s, 1H), 8.13 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 5.4, 1.2 Hz, 1H), 7.40 (dd, J = 3.6, 1.3 Hz, 1H), 7.23 - 7.18 (m, 2H), 7.08 (d, J = 1.7 Hz, 1H). MS (ESI + ): m / z = 289 ([M+H] + ).

[0094] Example 13: Synthesis of compound 2

[0095]

[0096] Synthesis method according to Example 12, white solid, 0.04 g, 36% yield.

[0097] 1 H NMR (600 MHz, chloroform-d) δ 10.37 (s, 1H), 8.47 (s, 1H), 8.14 (d, J = 8.5 Hz, 1H), 7.30 (dd, J = 8.5, 1.8 Hz, 1H), 7.24 (d, J = 1.8 Hz, 1H), 2.58 (s, 3H). MS (ESI + ): m / z = 221 ([M+H] + ).

[0098] Example 14: Synthesis of compound 3 and compound 4

[0099]

[0100] The reaction equation is as follows:

[0101]

[0102] Method A: Compound 1 (0.288 g, 1 mmol) was dissolved in dichloromethane (10 mL), 3-chloroperbenzoic acid (0.173 g, 1 mmol) was added, and stirred at 25 °C for 6 h. After the reaction was completed, sodium thiosulfate solution was added. The solution was separated, dried, filtered and concentrated, and purified by silica gel column chromatography to obtain white solid, compound 3, 0.28 g, yield 92%.

[0103] Method B: Compound 1 (0.288 g, 1 mmol) was dissolved in dichloromethane (2 mL), 3-chloroperbenzoic acid (0.173 g, 2 mmol) was added, and stirred at 30 °C for 1 h. After the reaction was completed, sodium thiosulfate solution was added. The solution was separated, dried, filtered and concentrated, and purified by silica gel column chromatography to obtain white solid, compound 3, 0.07 g, yield 23%, compound 4, 0.106 g, yield 33%. Compound 3: 1 HNMR (600 MHz, Chloroform-d) δ 10.37 (d, J = 1.5 Hz, 1H), 8.59 (s, 1H), 8.38 (d, J = 8.3 Hz, 1H), 8.06 (d, J = 1.6 Hz, 1H), 7.71 (dd, J = 3.7, 1.4 Hz, 1H), 7.69 (dd, J = 5.1, 1.3 Hz, 1H), 7.59 (dd, J = 8.3, 1.6 Hz, 1H), 7.13 (dd, J = 5.1, 3.7 Hz, 1H). MS (ESI + ): m / z = 305 ([M+H] + ). Compound 4: 1 HNMR (600 MHz, Chloroform-d) δ 10.35 (s, 1H), 8.59 (s, 1H), 8.43 (d, J = 8.3 Hz, 1H), 8.20 (d, J = 1.6 Hz, 1H), 8.03 (dd, J = 8.4, 1.6 Hz, 1H), 7.80 (dd, J = 3.8, 1.3 Hz, 1H), 7.76 (dd, J = 4.9, 1.3 Hz, 1H), 7.16 (dd, J = 5.0, 3.8 Hz, 1H). MS (ESI + ): m / z = 321 ([M+H] + ).

[0104] Example 15: Synthesis of compound 5

[0105]

[0106] Synthesis method refers to Example 12, light yellow solid, 0.118 g, yield 82%.

[0107] 1H NMR (600 MHz, chloroform-d) δ 10.34 (s, 1H), 8.47 (s, 1H), 8.21 (d, J = 8.4 Hz, 1H), 7.94 (d, J = 3.4 Hz, 1H), 7.53 (d, J = 3.4 Hz, 1H), 7.52 (d, J = 1.7 Hz, 1H), 7.44 (dd, J = 8.4, 1.7 Hz, 1H). MS (ESI + ): m / z = 290 ([M+H] + ).

[0108] Example 16: Synthesis of compound 6

[0109]

[0110] Synthesis method according to example 12, yellowish solid, 0.109 g, yield 70%.

[0111] 1 H NMR (600 MHz, chloroform-d) δ 10.39 (s, 1H), 8.55 (d, J = 5.6 Hz, 3H), 8.31 (d, J = 8.3 Hz, 1H), 7.92 (d, J = 1.5 Hz, 1H), 7.72 (dd, J = 8.3, 1.6 Hz, 1H), 7.10 (t, J = 4.8 Hz, 1H). MS (ESI + ): m / z = 285 ([M+H] + ).

[0112] Example 17: Synthesis of compound 7

[0113]

[0114] Synthesis method according to example 12, yellow solid, 0.095 g, yield 67%.

[0115] 1 H NMR (600 MHz, chloroform-d) δ 10.36 (s, 1H), 8.54 (ddd, J = 4.9, 2.0, 0.9 Hz, 1H), 8.49 (s, 1H), 8.22 (d, J = 8.3 Hz, 1H), 7.66 (td, J = 7.7, 1.9 Hz, 1H), 7.64 (d, J = 1.6 Hz, 1H), 7.51 (dd, J = 8.4, 1.7 Hz, 1H), 7.35 (dt, J = 8.0, 1.0 Hz, 1H), 7.21 (ddd, J = 7.5, 4.9, 1.1 Hz, 1H). MS (ESI + ): m / z = 284 ([M+H] + ).

[0116] Example 18: Synthesis of compound 8

[0117]

[0118] Synthesis method according to example 12, white solid, 0.105 g, yield 77%.

[0119] 1 H NMR (600 MHz, DMSO-d6) δ 10.09 (s, 1H), 8.84 (s, 1H), 8.04 (d, J = 8.4 Hz, 1H), 7.38 (s, 2H), 7.28 (d, J = 1.7 Hz, 1H), 7.19 (dd, J = 8.4, 1.8 Hz, 1H). MS (ESI + ): m / z = 273 ([M+H] + ).

[0120] Example 19: Synthesis of compound 9

[0121]

[0122] Synthesis method according to example 12, starting from product A-9 of example 9, the crude product was washed with ethanol and dried to obtain the product. Brown solid, 0.117 g, yield 46%.

[0123] 1 H NMR (600 MHz, DMSO-d6) δ 10.11 (s, 1H), 9.13 (s, 1H), 9.03 (d, J = 6.9 Hz, 2H), 8.95 (s, 1H), 8.58 (d, J = 6.9 Hz, 2H), 8.20 (d, J = 8.4 Hz, 1H), 8.08 (d, J = 1.8 Hz, 1H), 7.75 (dd, J = 8.4, 1.8 Hz, 1H), 4.34 (s, 3H), 3.17 (s, 1H). MS (ESI + ): m / z = 381 ([M-Cl - ] + ).

[0124] Example 20: Synthesis of compound 10

[0125]

[0126] Synthesis method according to example 12, yellow solid, 0.146 g, yield 86%.

[0127] 1H NMR (600 MHz, chloroform-d) δ 10.38 (s, 1H), 8.52 (s, 1H), 8.30 (d, J = 8.3 Hz, 1H), 8.01 (d, J = 8.2 Hz, 1H), 7.84 (d, J = 1.6 Hz, 1H), 7.81 (d, J = 8.0 Hz, 1H), 7.68 (dd, J = 8.3, 1.7 Hz, 1H), 7.51 (t, J = 7.1 Hz, 1H), 7.41 (t, J = 7.6 Hz, 1H). MS (ESI + ): m / z = 340 ([M+H] + ).

[0128] Example 21 : Synthesis of compound 11

[0129]

[0130] Synthesis method according to example 12, yellow solid, 0.135 g, yield 81%.

[0131] 1 H NMR (600 MHz, chloroform-d) δ 10.39 (s, 1H), 8.52 (s, 1H), 8.27 (d, J = 8.3 Hz, 1H), 8.08 (d, J = 8.6 Hz, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.84 (d, J = 1.6 Hz, 1H), 7.80 (d, J = 8.1 Hz, 1H), 7.75 - 7.69 (m, 1H), 7.65 (dd, J = 8.3, 1.5 Hz, 1H), 7.54 (t, J = 7.5 Hz, 1H), 7.32 (d, J = 8.6 Hz, 1H). MS (ESI + ): m / z = 356 ([M+Na] + ).

[0132] Example 22: Synthesis of compound 12

[0133]

[0134] Synthesis method according to example 12, yellow solid, 0.135 g, yield 81%.

[0135] 1H NMR (600 MHz, chloroform-d) δ 10.33 (s, 1H), 8.38 (s, 1H), 8.09 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 8.1 Hz, 1H), 7.94 (s, 1H), 7.74 (d, J = 8.4 Hz, 1H), 7.48 - 7.44 (m, 1H), 7.43 - 7.39 (m, 1H), 7.19 (dd, J = 8.5, 1.8 Hz, 1H), 6.99 (d, J = 1.8 Hz, 1H). MS (ESI + ): m / z = 361 ([M+Na] + ).

[0136] Example 23: Synthesis of compound 13

[0137]

[0138] Synthetic method according to example 12, yellow solid, 0.155 g, yield 83%.

[0139] 1 H NMR (600 MHz, chloroform-d) δ 10.35 (s, 1H), 8.45 (s, 1H), 8.13 (d, J = 8.4 Hz, 1H), 7.25 - 7.19 (m, 2H), 7.10 (s, 1H), 6.90 (d, J = 3.6 Hz, 1H), 2.95 (t, J = 7.6 Hz, 2H), 2.47 (t, J = 7.3 Hz, 2H), 2.11 - 2.03 (m, 2H). MS (ESI - ): m / z = 373 ([M-H] - ).

[0140] Example 24: Synthesis of compound 14 (as a comparative example)

[0141]

[0142] Chromone-3-carboxaldehyde (0.044 g, 0.25 mmol) was dissolved in tetrahydrofuran (2.5 mL), and aqueous sodium bisulfite solution (2 mol / L, 0.3 mmol) was added, and reacted at 30 °C for 1 hour. Filtration, the filter residue was washed with tetrahydrofuran and a small amount of ice water, and dried to obtain a white solid. 0.069 g, yield 99%.

[0143] 1H NMR (600 MHz, DMSO-d6) δ 8.38 (s, 1H), 8.07 (d, J = 8.0 Hz, 1H), 7.84 - 7.77 (m, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.5 Hz, 1H), 6.08 (d, J = 6.4 Hz, 1H), 5.33 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 255 ([M-Na] - ).

[0144] Example 25: Synthesis of compound 15

[0145]

[0146] Compound 1 was dissolved in tetrahydrofuran (2.5 mL), aqueous sodium bisulfite solution (2 mol / L, 0.3 mmol) was added, and the reaction was carried out at 30 °C for 1 h. Filtration, washing the filter residue with tetrahydrofuran and a small amount of ice water, and drying gave a white solid, 0.083 g, yield 85%. 1 H NMR (600 MHz, DMSO-d6) δ 8.38 (s, 1H), 8.07 (d, J = 8.0 Hz, 1H), 7.84 - 7.77 (m, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.5 Hz, 1H), 6.08 (d, J = 6.4 Hz, 1H), 5.33 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 255 ([M-Na] - ).

[0147] Example 26: Synthesis of compound 16

[0148]

[0149] Synthesis method according to Example 25, white solid, 0.075 g, yield 93%.

[0150] 1 H NMR (600 MHz, DMSO-d6) δ 8.38 (s, 1H), 8.07 (d, J = 8.0 Hz, 1H), 7.84 - 7.77 (m, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.50 (t, J = 7.5 Hz, 1H), 6.08 (d, J = 6.4 Hz, 1H), 5.33 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 255 ([M-Na] - ).

[0151] Example 27: Synthesis of compound 17

[0152]

[0153] Synthesis method according to example 25, white solid, 0.085 g, 83% yield.

[0154] 1 H NMR (600 MHz, DMSO-d6) δ 8.43 (s, 1H), 8.23 (d, J = 8.3 Hz, 1H), 8.00 (d, J = 7.7 Hz, 2H), 7.90 (d, J = 3.8 Hz, 1H), 7.71 (dd, J = 8.4, 1.7 Hz, 1H), 7.20 (t, J = 4.4 Hz, 1H), 6.18 (d, J = 6.6 Hz, 1H), 5.34 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 385 ([M-Na] - ).

[0155] Example 28: Synthesis of compound 18

[0156]

[0157] Synthesis method according to example 25, white solid, 0.094 g, 89% yield.

[0158] 1 H NMR (600 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.27 (d, J = 8.4 Hz, 1H), 8.24 (d, J = 1.9 Hz, 1H), 8.17 (dd, J = 4.9, 1.5 Hz, 1H), 8.01 - 7.96 (m, 2H), 7.28 (t, 1H), 6.10 (d, J = 6.5 Hz, 1H), 5.30 (d, J = 6.3 Hz, 1H). MS (ESI - ): m / z = 401 ([M-Na] - ).

[0159] Example 29: Synthesis of compound 19

[0160]

[0161] Synthesis method according to example 25, white solid, 0.087 g, 89% yield.

[0162] 1H NMR (600 MHz, DMSO-d6) δ 8.35 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.99 (d, J = 3.4 Hz, 1H), 7.95 (d, J = 3.5 Hz, 1H), 7.72 (d, J = 1.8 Hz, 1H), 7.48 (dd, J = 8.4, 1.9 Hz, 1H), 6.10 (d, J = 6.4 Hz, 1H), 5.31 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 370 [M-Na] - ).

[0163] Example 30: Synthesis of compound 20

[0164]

[0165] Synthesis method according to example 25, white solid, 0.076 g, yield 81%.

[0166] 1 H NMR (600 MHz, DMSO-d6) δ 8.35 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.99 (d, J = 3.4 Hz, 1H), 7.95 (d, J = 3.5 Hz, 1H), 7.72 (d, J = 1.8 Hz, 1H), 7.48 (dd, J = 8.4, 1.9 Hz, 1H), 6.10 (d, J = 6.4 Hz, 1H), 5.31 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 370 [M-Na] - ).

[0167] Example 31: Synthesis of compound 21

[0168]

[0169] Synthesis method according to example 25, white solid, 0.076 g, yield 81%.

[0170] 1 H NMR (600 MHz, DMSO-d6) δ 8.35 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.99 (d, J = 3.4 Hz, 1H), 7.95 (d, J = 3.5 Hz, 1H), 7.72 (d, J = 1.8 Hz, 1H), 7.48 (dd, J = 8.4, 1.9 Hz, 1H), 6.10 (d, J = 6.4 Hz, 1H), 5.31 (d, J = 6.5 Hz, 1H). MS (ESI - ): m / z = 370 [M-Na]- ).

[0171] Example 32: Synthesis of compound 22

[0172]

[0173] Synthesis method according to example 25, white solid, 0.092 g, yield 84%.

[0174] 1 H NMR (600 MHz, DMSO-d6) δ 8.40 (s, 2H), 8.30 (d, J = 8.7 Hz, 2H), 8.13 (d, J = 8.3 Hz, 2H), 8.03 - 7.92 (m, 4H), 7.86 (d, J = 8.5 Hz, 2H), 7.75 (t, J = 7.7 Hz, 2H), 7.67 (d, J = 8.3 Hz, 2H), 7.58 (t, J = 7.5 Hz, 2H), 7.37 (d, J = 8.6 Hz, 2H), 6.18 (d, J = 6.6 Hz, 2H), 5.37 (d, J = 6.4 Hz, 2H). MS (ESI - ): m / z = 414 [M-Na] - ).

[0175] Example 33: Synthesis of compound 23

[0176]

[0177] Synthesis method according to example 25, white solid, 0.088 g, yield 80%.

[0178] 1 H NMR (600 MHz, DMSO-d6) δ 8.47 (s, 1H), 8.24 (s, 1H), 8.16 (d, J = 8.0 Hz, 1H), 7.93 (d, J = 8.4 Hz, 1H), 7.64 (d, J = 7.9 Hz, 1H), 7.49 (t, J = 7.5 Hz, 1H), 7.44 (t, J = 7.5 Hz, 1H), 7.16 (d, J = 8.5 Hz, 1H), 7.13 (s, 1H), 6.08 (d, J = 6.5 Hz, 1H), 5.29 (d, J = 6.3 Hz, 1H). MS (ESI - ): m / z = 419 [M-Na] - ).

[0179] Example 34: Synthesis of compound 24

[0180]

[0181] Synthetic procedure according to Example 25, white solid, 0.096 g, 80% yield.

[0182] 1 H NMR (600 MHz, DMSO-d6) δ 8.28 (s, 1H), 7.97 (d, J = 8.3 Hz, 1H), 7.40 (d, J = 3.5 Hz, 1H), 7.23 - 7.17 (m, 2H), 7.03 (d, J = 3.6 Hz, 1H), 6.01 (s, 1H), 5.27 (s, 1H), 2.88 (t, J = 7.6 Hz, 2H), 2.28 (t, J = 7.3 Hz, 2H), 1.91 - 1.83 (m, 2H). MS (ESI - ): m / z = 455 [M-Na] - ).

[0183] The compounds synthesized according to Examples 12-34 of the present application were tested for their inhibitory effect on NDM-1 and the corresponding half-inhibitory concentrations IC 50 values (as shown in Table 1) were obtained. As can be seen from the results in Table 1, the inhibitory activity of chromone-3-carboxaldehyde (IC 50 = 4.07 μΜ) is essentially the same as that of the corresponding derivative 14 (IC 50 = 4.08 μΜ), which indicates that derivatization does not affect the activity of the aldehyde group. Therefore, only the activity of the corresponding derivatives of compounds 1-5, 8, 10-13 was tested in Table 1. Except for compound 18, the synthesized chromone-3-carboxaldehyde derivatives have different degrees of enhancement of inhibitory effect on NDM-1, among which the IC 50 of compound 23 is 0.037 μΜ, which is about 1000-fold and 100-fold higher than the inhibitory activity of positive control captopril (IC 50 = 33.2 μΜ) and the parent compound 14, respectively, and can reduce the MIC of imipenem and meropenem against NDM-1-producing E. coli by 16 and 64 times, respectively (as shown in Table 2; the MIC test was performed according to the requirements of the American Clinical and Laboratory Standards Association (CLSI): E. coli BL21 (DE3)-pET15b (+)-blaNDM-1 (final concentration of 5 x 10 5CFU / mL) were mixed with imipenem (final concentration 0-128 mg / L) or meropenem (final concentration 0-128 mg / L) in cation-adjusted Muller-Hinton broth, and the tested compounds (final concentration 16 mg / L) were added. After incubation at 37°C for 20 hours, the changes of MIC of imipenem or meropenem were observed. It was shown that the synthesized chromone compounds were excellent NDM-1 inhibitors, and by combination with β-lactam antibiotics, they were expected to treat diseases caused by NDM-1 expressing drug-resistant bacterial infections, and had good prospects for drug application.

[0184] Table 1

[0185]

[0186]

[0187] Table 2

[0188]

Claims

1. A 7-sulfur-containing substituent-chromone-3-carboxaldehyde compound represented by Formula I or a compound represented by Formula II or a compound represented by Formula 16: ###0001### ###0002### ###0003### wherein, R is selected from the group consisting of thiazol-2-yl, pyrimidin-2-yl, pyridin-2-yl, imidazol-2-yl, (1-methyl-pyridin-4)yl-4-thiazol-2-yl, benzothiazol-2-yl, quinolin-2-yl, benzothiophen-2-yl, (4-carboxypropyl)-5-thiophen-2-yl, n represents the number of S to which R is attached; when R is selected from thien-2-yl, n is selected from 0, 1 or 2; when R is selected from groups other than thien-2-yl, n is selected from 0. R is selected from the group consisting of thiazol-2-yl, pyrimidin-2-yl, pyridin-2-yl, imidazol-2-yl, (1-methyl-pyridin-4)yl-4-thiazol-2-yl, benzothiazol-2-yl, quinolin-2-yl, benzothiophen-2-yl, (4-carboxypropyl)-5-thiophen-2-yl, n represents the number of S to which R is attached; when R is selected from thien-2-yl, n is selected from 0, 1 or 2; when R is selected from groups other than thien-2-yl, n is selected from 0.

2. The process for the preparation of 7-sulpho-substituted-chromone-3- carbaldehydes of formula I according to claim 1, characterized in that, when n is 0, the 7-sulfur-substituted-chromone-3-carboxaldehyde compound of formula I has the following structure The preparation method comprises the following steps: taking 4-sulfur-substituted-2-hydroxyacetophenone as a raw material, reacting in a POCl3 and N,N-dimethylformamide system at 25-40°C for 6-18 hours to obtain the target 7-sulfur-substituted-chromone-3-carboxaldehyde compound; the molar ratio of the compound A and POCl3 is 1:1.5-5, and the reaction formula is as follows:

3. The method of claim 2, wherein, The preparation method of compound A is as follows: 4-mercapto-2-hydroxyacetophenone, the aromatic iodide, potassium hydroxide and cuprous oxide are in a molar ratio of 1:(1-2):(1.5-5):(0.02-0.2); the volume ratio of dimethyl sulfoxide and water in the mixed solvent is 4:1-0.2, and the reaction formula is as follows: or, 4-mercapto-2-hydroxyacetophenone, the aromatic iodide, potassium hydroxide and cuprous oxide are in a molar ratio of 1:(1-2):(1.5-5):(0.02-0.2); the volume ratio of dimethyl sulfoxide and water in the mixed solvent is 4:1-0.2; and the reaction formula is as follows:

4. The method of claim 3, wherein, The concentration of 4-mercapto-2-hydroxyacetophenone is 0.1-1 mol / L; and the concentration of 4-iodo-2-hydroxyacetophenone is 0.2-2 mol / L.

5. The method of claim 2, wherein, When the R substituent in compound A is thiazole-2-yl, the preparation method of compound A-1 is as follows: 4-fluoro-2-hydroxyacetophenone and thiophene-2-thiol are used as raw materials, potassium carbonate is used as a base, and 4-(thiophene-2-thio) hydroxyacetophenone is obtained by reacting at 90-130 °C in N,N-dimethylformamide solvent for 1-6 hours; the molar ratio of 4-fluoro-2-hydroxyacetophenone, thiophene-2-thiol and potassium carbonate is 1:(1-2):(1-2); and the reaction formula is as follows:

6. The method of claim 5, wherein, The concentration of 4-fluoro-2-hydroxyacetophenone is 0.1-1 mol / L.

7. The method of claim 2, wherein, When the R substituent in Compound A is (1-methyl-pyridin-4)yl-4-thiazol-2-yl, the compound is prepared according to the following scheme:

8. The process for the preparation of 7-sulpho-substituted-chromone-3- carbaldehydes of formula I according to claim 1, characterized in that, When n is selected from 1 or 2, 7-(thiophene-2-thio) chromone-3-carboxaldehyde obtained by the preparation method in claim 2 is used as raw material, 3-chloroperbenzoic acid is used as a solvent, and dichloromethane is used as a solvent, and compound B is obtained by reacting at 20-30 °C for 1-6 hours; the molar ratio of compound B and 3-chloroperbenzoic acid is 1:1-2; and the reaction formula is as follows:

9. A process for the preparation of a compound of formula II as claimed in claim 1, characterized in that, 7-sulfur-containing substituent-chromone-3-carboxaldehyde compounds represented by Formula I are used as raw materials, an aqueous sodium bisulfite solution is used as a solvent, and the reaction is carried out at 20-40 °C for 0.5-4 hours in tetrahydrofuran to obtain compounds represented by Formula II; the molar ratio of 7-sulfur-containing substituent-chromone-3-carboxaldehyde compounds and sodium bisulfite is 1:1-2.

5.

10. Use of the 7-sulfur-containing substituent-chromone-3-carboxaldehyde compound represented by Formula I or the compound represented by Formula II or the compound represented by Formula 16 in claim 1 as an inhibitor of New Delhi metallo-beta-lactamase NDM-1 in the preparation of a drug for inhibiting the activity of NDM-1.

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