2-oximin-1h-quinoline-4-carboxylic acid derivatives, process for their preparation and use thereof

Abstract

The application discloses a 2-oxo-1H-quinoline-4-carboxylic acid derivative and a preparation method and application thereof. The 2-oxo-1H-quinoline-4-carboxylic acid derivative has a compound structure as shown in a general formula (I). The 2-oxo-1H-quinoline-4-carboxylic acid derivative can effectively inhibit NQO1 activity, can be combined with an antitumor drug, and can be used for treating pancreatic cancer and non-small cell lung cancer, while reducing a dosage of the antitumor drug, reducing side effects, and ensuring medication safety.

Description

Technical Field

[0001] This invention relates to NQO1 inhibitors, and more particularly to a 2-oxo-subunit-1H-quinoline-4-carboxylic acid derivative, its preparation method and application, belonging to the field of compound pharmaceuticals. Background Technology

[0002] NAD(P)H:quinone oxidoreductase 1 (NQO1) is a homodimeric flavinase, also known as D-lipoamide dehydrogenase (DT-diaphorase). It is a widely distributed two-electron oxidoreductase in cells, primarily located in the cytoplasm (>90%) and nucleus. Using flavin adenine dinucleotide (FAD) as a cofactor and coenzyme NAD(P)H as an electron donor, it reduces quinones to hydroquinones via a one-step two-electron transfer process. The two-electron reduction of substrates catalyzed by NQO1 is accomplished through a "ping-pong mechanism." As a phase II metabolic enzyme, NQO1 is not abundantly metabolized in normal human tissues. However, under oxidative stress, NQO1 is transcribed and expressed under the control of relevant signaling pathways, exerting its physiological functions. The main functions include: 1) performing two-electron reduction on quinones and their derivatives to prevent them from being activated by single-electron reductases to produce semiquinone free radicals and ROS, thereby preventing quinones from directly harming the human body; 2) serving as a component of the body's antioxidant system by directly scavenging superoxide anions through NAD(P)H-dependent reduction reactions; 3) maintaining the reduced state of endogenous lipid-soluble antioxidants α-tocopherol and ubiquinone to preserve their activity; and stabilizing tumor suppressor p53 and other proteins through protein-protein interactions.

[0003] Studies have shown that the NQO1 enzyme is upregulated in many solid tumors, with its expression level in over 80% of human pancreatic cancers being 100 times higher than in normal tissues. Besides lung and pancreatic cancer, NQO1 expression levels in other solid tumors, including breast, prostate, and colorectal cancers, are significantly higher than in normal tissues. The promoting effect of NQO1 enzyme on tumorigenesis and development, along with its high expression in tumor tissues, indicates that this protein is a highly selective target for anti-tumor therapy. Research has reported that NQO1 overexpression induced by the transcription factor NRF2 may lead to chemotherapy failure, possibly as an adaptive response to oxidative stress and cytotoxicity, and as a defense mechanism for cancer cells. Therefore, inhibiting the NQO1 enzyme could enable selective anti-cancer therapy addressing chemotherapy failure caused by NQO1-catalyzed detoxification. Furthermore, inhibiting NQO1 enzyme expression increases intracellular ROS in tumor cells, increasing the risk of anodic apoptosis and reducing tumor cell invasiveness, thus exploring new therapeutic directions for cancer treatment. Meanwhile, NQO1 enzyme inhibitors can also provide valuable assistance for in-depth research on the pharmacological function of the NQO1 enzyme. Currently, some compounds are known to competitively inhibit NQO1 activity by binding to NAD(P)H, thereby preventing the reduction of FAD, including guar gum, coumarin, curcumin, triazolidine-6-one, etc., with dicumarin (DIC) being the most representative. DIC is frequently used as an NQO1 inhibitor to study the function of NQO1 in cells. However, this drug also has certain drawbacks; the effect of dicumarin is affected by extensive protein binding and confounding "off-target" effects, such as mitochondrial uncoupling and increased intracellular superoxide production. Therefore, there is still a need to develop NQO1 inhibitors with better efficacy, and this invention provides such compounds. Summary of the Invention

[0004] Objectives of the invention: The objective of this invention is to provide a compound or a pharmaceutically acceptable salt thereof that can effectively inhibit NQO1 activity; another objective of this invention is to provide a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof; another objective of this invention is to provide a pharmaceutical composition; and another objective of this invention is to provide an application of the compound or a pharmaceutically acceptable salt thereof.

[0005] Technical solution: A compound of general formula (Ⅰ) or a pharmaceutically acceptable salt thereof, according to the present invention.

[0006]

[0007] Among them, Ar 1 Represents a 5- to 6-membered aromatic ring containing carbon and / or nitrogen atoms;

[0008] Ar 2 Represents a benzene ring, triazole, or a 5- to 6-membered aromatic heterocycle;

[0009] Ar 3 It represents an aryl group containing 5 to 6 atoms, or an 8 to 10-membered bicyclic aryl or heteroaryl group, wherein a heteroaryl group is an aryl group containing 1 to 2 identical or different heteroatoms selected from nitrogen and oxygen in the aromatic ring member;

[0010] R 1 Represents a hydrogen atom or located in Ar 1 One or more substituents at any position on the ring, wherein the substituents are selected from C1-C4 alkyl, C1-C4 alkoxy, halogen, acyl, and hydroxyl groups;

[0011] R 2 This indicates that the atom does not exist, the hydrogen atom, the methylene group, or the atom located in Ar. 2 One or more substituents at any position on the ring, wherein the substituents are selected from hydrogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkylamino, C1-C4 haloalkyl, halogen, trifluoromethoxy, cyano, hydroxy, amino, carboxyl, and nitro.

[0012] R 3 Representative located in Ar 3 One or more substituents at any position on the ring, wherein the substituents are selected from hydrogen, alkyl, C1-C4 alkoxy, C1-C4 alkylamino, C1-C4 haloalkyl, halogen, phenyl, trifluoromethoxy, cyano, hydroxy, amino, carboxyl, aminosulfonyl, C1-C4 alkylsulfonyl, C1-C4 amide, and C1-C4 alkylaminoyl.

[0013] When m is 0, X represents CH2; when m is 1, X represents O.

[0014] In general formula (I), the dashed line represents the Ar group shown by the dashed line. 3 and R 3 There are two possibilities: presence or absence. Preferably, the above-described compound or its pharmaceutically acceptable salt has a chemical structure as shown in (I-a) or (I-b):

[0015]

[0016] Among them, R 2 The meaning of "representing non-existence" is that Ar does not exist. 2 and Ar 3 Direct connection, i.e.

[0017] Furthermore, Ar 1 Represents benzene ring, pyridine; Ar 2 Represents a benzene ring, triazole, or a 5- to 6-membered aromatic heterocycle containing 1-3 nitrogen atoms; Ar 3Representing benzene rings, 5-6 membered aromatic heterocycles, and 8-10 membered bicyclic aryl groups, wherein the 5-6 membered aromatic heterocycle is an aryl group containing 1-2 identical or different heteroatoms selected from nitrogen and oxygen.

[0018] Preferably, the 8- to 10-membered bicyclic aryl group is a fused ring or biphenyl.

[0019] Furthermore, R 1 Represents H, -CH3, -OCH3, -F, -OC(O)CH3, and -OH;

[0020] R 2 The following groups are not represented: -Cl, -F, trifluoromethoxy, nitro, -CH3, -CH2NH2, -OCF3, -COOH, -CN, methoxy, -OH, SO2CH3, -C(O)NH2, C(O)NHCH2CH3.

[0021] Furthermore,

[0022] Ar 1 Selected from

[0023] Ar 2 Selected from Ar 3 Selected from

[0024] Furthermore,

[0025] Ar 1 Selected from Ar 2 Selected from Ar 3 Selected from Typical compounds of this invention include, but are not limited to:

[0026]

[0027]

[0028]

[0029]

[0030]

[0031]

[0032]

[0033] Preferably, compounds of general formula (I) may contain acidic functional groups sufficient to form salts. Representative salts include pharmaceutically acceptable metal salts such as sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc salts; pharmaceutically acceptable metal cations such as carbonates and bicarbonates of sodium, potassium, lithium, calcium, magnesium, aluminum, and zinc; and pharmaceutically acceptable primary, secondary, and tertiary organic amines, including aliphatic amines, aromatic amines, aliphatic diamines, and hydroxyalkyl amines, such as methylamine, ethylamine, 2-hydroxyethylamine, diethylamine, triethylamine, ethylenediamine, ethanolamine, and diethanolamine.

[0034] On the other hand, the present invention provides a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof.

[0035]

[0036] ;or,

[0037]

[0038]

[0039] Among them, R 1 R 2 R 3 Ar 1 Ar 2 Ar 3 The definitions of X and m are as described above.

[0040] On the other hand, the present invention provides a pharmaceutical composition comprising the above-described compound or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0041] On the other hand, the present invention provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating diseases related to NQO1 enzyme abnormalities.

[0042] Preferably, diseases associated with NQO1 enzyme abnormalities include pancreatic cancer and non-small cell lung cancer.

[0043] On the other hand, the present invention provides the use of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of NQO1 inhibitors.

[0044] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: it can effectively inhibit the activity of NQO1 by 2-oxo-1H-quinoline-4-carboxylic acid derivatives, which can be used in combination with anti-tumor drugs to treat pancreatic cancer and non-small cell lung cancer, while reducing the dosage of anti-tumor drugs, reducing side effects, and ensuring drug safety. Detailed Implementation

[0045] Example 1: Preparation of compound I-1:

[0046] 7-Fluoro-1H-indole-2,3-dione (330 mg, 2 mmol) was dissolved in 20 mL of ethanol, followed by the addition of ethyl diazonate (420 μL, 4 mmol) and DBU (60 μL, 0.3 mmol). The reaction solution was placed at 40 °C for 15 h. After the reaction was completed, the solvent was evaporated to dryness, and 20 mL of 1 N hydrochloric acid was added. A solid precipitated out, which was filtered to obtain a pale yellow solid (382 mg, 76%). Ethyl 8-fluoro-3-hydroxy-2-oxoylide-1H-quinoline-4-carboxylate (200 mg, 0.8 mmol) was dissolved in 15 mL of DMF, followed by the addition of benzyl bromide (110 μL, 0.96 mmol) and anhydrous potassium carbonate (220 mg, 1.6 mmol). The reaction solution was placed at 66 °C for 10 h. After the reaction was complete, 20 mL of water and ethyl acetate were added for extraction. The organic phase was retained, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography with petroleum ether / ethyl acetate (6:1) to give a white solid (101 mg, 37.4%). Ethyl 2-(benzyloxy)-8-fluoro-2-oxoylide-1H-quinoline-4-carboxylate (70 mg, 0.2 mmol) was dissolved in 10 mL of a 1:1 mixture of methanol and tetrahydrofuran. Potassium hydroxide (460 mg, 8.2 mmol) and 5 mL of water were then added, and the reaction mixture was placed at 65 °C for 18 h. After the reaction was complete, the organic solvent was evaporated to dryness, and the pH was adjusted to 2 with 1 N hydrochloric acid. A white solid precipitated, which was filtered and dried to give 56.3 mg of white solid, with a total yield of 9.0%. 1 HNMR (400MHz, DMSO-d6) δ14.12(s,1H),12.35(s,1H),7.50-7.45(m,2H),7.44-7.37(m,3H),7.37-7.33(m,1H),7.26-7.20(m,2H),5.28(s,2H); EI-MS m / z:312.08[M] - .

[0047] Example 2: Preparation of compound I-2:

[0048] The method in Example 2 was the same as in Example 1, except that benzyl chlorobromobenzyl (0.96 mmol) was used instead of benzyl bromobenzyl, yielding 73.4 mg of a white solid, with an overall yield of 10.6%. 1 H NMR (400MHz, DMSO-d6) δ14.15(s,1H),12.36(s,1H),7.51-7.44(m,4H),7.44-7.38(m,1H),7.26-7.20(m,2H),5.27(s,2H); EI-MS m / z:346.01[M] - .

[0049] Example 3: Preparation of compound I-3:

[0050] The method in Example 3 was the same as in Example 1, except that benzyl bromo ... 1 EI-MS m / z:330.14[M] - .

[0051] Example 4: Preparation of compound I-4:

[0052] The method in Example 4 was the same as in Example 1, except that 7-fluoro-1H-indole-2,3-dione was replaced with 7-methoxy-1H-indole-2,3-dione (2 mmol), yielding 64.2 mg of a white solid, with an overall yield of 9.8%. 1 H NMR(400MHz,DMSO-d6)δ13.93(s,1H),11.39(s,1H),7.50-7.45(m,2H),7.42-7.31(m ,3H),7.23-7.12(m,2H),6.98(dd,J=7.9,1.3Hz,1H),5.26(s,2H),3.92(s,3H); EI-MS m / z:324.32[M] - .

[0053] Example 5: Preparation of compound I-5:

[0054] The method in Example 5 was the same as in Example 1, except that 7-fluoro-1H-indole-2,3-dione (2 mmol) was replaced with 7-methoxy-1H-indole-2,3-dione, and benzyl bromo ... 1 HNMR (400MHz, DMSO-d6) δ13.95(s,1H),11.41(s,1H),7.47(q,J=8.5Hz,4H),7.22-7.12(m,2H),6.98(d,J=7.8Hz,1H),5.25(s,2H),3.92(s,3H); EI-MS m / z:358.4[M] - .

[0055] Example 6: Preparation of compound I-6:

[0056] The method in Example 6 was the same as in Example 1, except that 7-fluoro-1H-indole-2,3-dione (2 mmol) was replaced with 7-methoxy-1H-indole-2,3-dione, and benzyl bromo ... 1 HNMR(400MHz,DMSO-d6)δ13.94(s,1H),11.41(s,1H),7.51(dd,J=8.5,5.8Hz, 2H),7.26-7.11(m,4H),6.98(d,J=8.0Hz,1H),5.24(s,2H),3.92(s,3H); EI-MS m / z:342.3[M] - .

[0057] Example 7: Preparation of compound I-7:

[0058] The method in Example 7 was the same as in Example 1, except that 7-fluoro-1H-indole-2,3-dione was replaced with 7-hydroxy-1H-indole-2,3-dione (2 mmol), yielding 88.4 mg of a white solid, with an overall yield of 14.2%. 1 H NMR(300MHz,DMSO-d6)δ13.83(s,1H),11.15(s,1H),10.45(s,1H),7.48-7.35(m,5H), 7.06(t,J=7.9Hz,1H),6.93(d,J=6.6Hz,1H),6.85(d,J=8.0Hz,1H),5.24(s,2H); EI-MS m / z:310.3[M] - .

[0059] Example 8: Preparation of compound I-8:

[0060] The method in Example 8 was the same as in Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and benzyl bromo ... 1 HNMR(400MHz,DMSO-d6)δ13.89(s,1H),11.12(s,1H),10.43(s,1H),7.47(q,J=8.6Hz,4H),7.06 (t,J=7.9Hz,1H),6.94(d,J=6.7Hz,1H),6.85(d,J=7.3Hz,1H),5.24(s,2H);EI-MSm / z:344.8[M] - .

[0061] Example 9: Preparation of compound I-9:

[0062] The method of Example 9 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and benzyl bromo ... 1 HNMR(400MHz,DMSO-d6)δ13.86(s,1H),11.11(s,1H),10.42(s,1H),7.51(dd,J=8.5,5.8Hz,2H),7.21( t,J=8.9Hz,2H),7.06(t,J=7.9Hz,1H),6.93(d,J=7.9Hz,1H),6.85(d,J=8.0Hz,1H),5.23(s,2H); EI-MS m / z:328.3[M] - .

[0063] Example 10: Preparation of compound I-10:

[0064] The method of Example 10 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and benzyl bromo ... 1 H NMR (400MHz, DMSO-d6) δ13.43(s,1H),10.34(d,J=5.2Hz,2H),7.70-7.63(m,2H),7.50(dt,J=7.1,1.0Hz,2H EI-MS m / z:378.3[M] - .

[0065] Example 11: Preparation of compound I-11:

[0066] The method of Example 11 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and 1-(bromomethyl)-4-nitrobenzene (0.96 mmol) was replaced with benzyl bromo, to obtain 88.5 mg of white solid, with an overall yield of 12.9%. 1H NMR(400MHz,Chloroform-d)δ13.06(s,1H),9.84(s,1H),8.71(s,1H),8.14-8.07(m,2H),7.66-7.58(m,2H EI-MS m / z:355.1[M] - .

[0067] Example 12: Preparation of compound I-12:

[0068] The method of Example 12 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and 4-methylbenzyl bromide (0.96 mmol) was replaced with benzyl bromide, to obtain 76.3 mg of white solid, with an overall yield of 11.7%. 1 H NMR (400MHz, DMSO-d6) δ13.43(s,1H),10.34(d,J=5.2Hz,2H),7.46(dd,J=8.1,1.2Hz,1H),7.31-7.21(m,1H EI-MS m / z:324.32[M] - .

[0069] Example 13: Preparation of compound I-13:

[0070] The method of Example 13 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and [4-(bromomethyl)phenyl]methaneamine (0.96 mmol) was replaced with benzyl bromo, to obtain 99.3 mg of white solid, with an overall yield of 14.6%. 1 H NMR(400MHz, DMSO-d6)δ13.43(s,1H),10.34(d,J=5.2Hz,2H),7.46(dd,J=8.1,1.2Hz,1H),7.36-7.22(m,5H), 6.94(dd,J=8.7,1.2Hz,1H),5.30(t,J=1.0Hz,2H),3.97(tt,J=6.2,1.0Hz,2H),3.45(d,J=12.6Hz,1H); EI-MS m / z:339.34[M]- .

[0071] Example 14: Preparation of compound I-14:

[0072] The method in Example 14 was the same as in Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and 1-(bromomethyl)-3-[(trifluoromethyl)oxy]benzene (0.96 mmol) was replaced with bromobenzyl, yielding 99.9 mg of a white solid with an overall yield of 12.6%. 1 H NMR (400MHz, DMSO-d6) δ13.43(s,1H),10.34(d,J=5.2Hz,2H),7.46(dd,J=8.1,1.2Hz,1H),7.31-7.22( EI-MS m / z:394.35[M] - .

[0073] Example 15: Preparation of compound I-15:

[0074] The method of Example 15 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and 5-(bromomethyl)pyridine-2-carboxylic acid (0.96 mmol) was replaced with benzyl bromo, to obtain 82.3 mg of white solid, with an overall yield of 11.5%. 1 H NMR (400MHz, DMSO-d6) δ13.43(s,1H),12.82(s,1H),10.34(d,J=5.2Hz,2H),8.59(d,J=2.2Hz,1H),8.06(d,J=8.1Hz,1H),7 .69(dd,J=8.1,2.3Hz,1H),7.46(dd,J=8.1,1.2Hz,1H),7.31-7.22(m,1H),6.94(dd,J=8.7,1.2Hz,1H),5.46(s,1H); EI-MS m / z:355.29[M] - .

[0075] Example 16: Preparation of compound I-16:

[0076] The method of Example 16 was the same as that of Example 1, except that 7-fluoro-1H-indole-2,3-dione was replaced with 6-methyl-1H-pyrrolo[2,3-b]pyridine-2,3-dione (2 mmol), and bromobenzyl was replaced with 4-(bromomethyl)aniline (0.96 mmol), to obtain 82.3 mg of white solid, with an overall yield of 11.5%. 1H NMR(400MHz,Chloroform-d)δ13.12(s,1H),9.87(s,1H),7.85(d,J=8.2Hz,1H),7.32-7.23(m,2H),7.17- 7.10(m,1H),6.61-6.54(m,2H),5.30(t,J=0.9Hz,2H),4.17(d,J=5.7Hz,1H),4.07(d,J=5.7Hz,1H); EI-MS m / z:324.11[M] - .

[0077] Example 17: Preparation of compound I-17:

[0078] 7-hydroxy-1H-indole-2,3-dione (326 mg, 2 mmol) was dissolved in 10 mL of DMF. The reaction solution was placed in an ice bath at 0 °C, and then NaH (48 mg, 2 mmol) was added. After stirring for 10 min, acetic anhydride (281 μL, 3 mmol) was added, and the reaction was allowed to proceed for 2 h. After the reaction was completed, 20 mL of water was added to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL × 3), retaining the organic phase, and dried over anhydrous sodium sulfate. The solution was purified by silica gel column chromatography with petroleum ether / ethyl acetate (5:1) to give a yellow solid (390 mg, 95.1%), namely acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester. The method in Example 17 was the same as in Example 1, except that 7-fluoro-1H-indole-2,3-dione was replaced with 2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (2 mmol), and benzyl bromoacetate was replaced with 4-(bromomethyl)benzyl-1-carboxynitrile (0.96 mmol), yielding 105.3 mg of a white solid, with an overall yield of 13.9%. 1 H NMR(400MHz,DMSO-d6)δ13.27(s,1H),10.54(s,1H),7.70-7.63(m,2H),7.53-7.46( m,2H),7.46-7.38(m,2H),7.21(dd,J=6.0,4.2Hz,1H),5.30(t,J=1.0Hz,2H); EI-MS m / z:377.34[M] - .

[0079] Example 18: Preparation of compound I-18:

[0080] The method of Example 18 was the same as that of Example 1, except that 7-hydroxy-1H-indole-2,3-dione (2 mmol) was replaced with 7-fluoro-1H-indole-2,3-dione, and benzyl bromo ... 1 H NMR(400MHz,Chloroform-d)δ13.06(s,1H),9.84(s,1H),8.71(s,1H),7.45(dd,J=8.2,1.3Hz,1H), 7.38-7.30(m,2H),7.30-7.22(m,1H),6.98-6.88(m,3H),5.30(t,J=1.0Hz,2H),3.78(s,3H); EI-MS m / z:340.3[M] - .

[0081] Example 19: Preparation of compound I-19:

[0082] 1-(azidomethyl)-4-fluorobenzene (1 g, 5 mmol) was dissolved in 5 mL of anhydrous DMF, followed by the addition of pentyl-4-acetylic acid (490 mg, 5 mmol), copper sulfate pentahydrate (62.5 mg, 0.25 mmol), and sodium ascorbate (49.5 mg, 0.25 mmol). The reaction mixture was placed at 80 °C for 15 h. After the reaction was completed, the mixture was cooled to room temperature, extracted with water and ethyl acetate, and the organic phase was retained. The organic phase was dried over anhydrous sodium sulfate and purified by silica gel column chromatography with petroleum ether / ethyl acetate (1:1) to give a white solid (1.3 g, 87.6%). 3-{1-[(4-fluorophenyl)methyl]-1,2,3-triazacyclopentan-4-yl}propionic acid (460 mg, 1.85 mmol) was placed in a 50 mL reaction flask, and 3 mL of thionyl chloride was slowly added dropwise. The mixture was stirred at room temperature for 2 h, and then the thionyl chloride was removed by rotary evaporation to obtain a light yellow oil (0.49 g, 98.9%). 7-Methoxy-1H-indole-2,3-dione (177 mg, 1 mmol) was dissolved in 10 mL of dichloromethane, followed by the addition of pyridine (320 mL, 4 mmol). Then, 5 mL of a dichloromethane solution of 3-{1-[(4-fluorophenyl)methyl]-1,2,3-triazacyclopentanyl-4-yl}propionyl chloride (350 mg, 1.33 mmol) was added dropwise. After the addition was complete, the reaction solution was placed at 50 °C for 12 h. The reaction was then stopped, cooled to room temperature, washed with 1 N hydrochloric acid, and extracted three times with dichloromethane. The organic phase was retained, dried over anhydrous sodium sulfate, and purified by dichloromethane / methanol (50:1) silica gel column chromatography to give a brown solid (110 mg, 31.2%). 1-(3-{1-[(4-fluorophenyl)methyl]-1,2,3-triazacyclopentanyl-4-yl}propionyl)-7-methoxyindole-2,3-dione (180 mg, 0.43 mmol) was dissolved in 15 mL of a mixture of methanol and water (2:1), followed by the addition of sodium hydroxide (185 mg, 4.3 mmol). The reaction mixture was placed at 105 °C for 12 h. After the reaction was completed, the mixture was cooled to room temperature, the methanol was removed by rotary evaporation, and 1 N hydrochloric acid was added to adjust the pH to 2. A white solid precipitated out. The solid was filtered and dried to obtain 150 mg of white solid, with an overall yield of 36.7%. 1 H NMR(300MHz,DMSO-d6)δ14.14(s,1H),11.20(s,1H),7.76(s,1H),7.36-7.31(m,2H ),7.21-7.14(m,4H),7.08-7.02(m,1H),5.49(s,2H),3.91(d,J=1.8Hz,5H); EI-MS m / z:407.1[M] - .

[0083] Example 20: Preparation of compound I-20:

[0084] The method of Example 20 was the same as that of Example 19, except that 1-(azidomethyl)-4-(trifluoromethyl)benzene (5 mmol) was replaced with 1-(azidomethyl)-4-fluorobenzene, yielding 110.3 mg of white solid, with an overall yield of 24.1%. 1 H NMR (400MHz, DMSO-d6) δ14.18(s,1H),11.18(s,1H),7.83(s,1H),7.72(d,J=8.0Hz,2H),7.44(d,J=8.0Hz, 2H),7.19(d,J=5.3Hz,2H),7.05(p,J=4.2Hz,1H),5.63(s,2H),3.92(d,J=6.4Hz,5H);EI-MSm / z:457.1[M] - .

[0085] Example 21: Preparation of compound I-21:

[0086] The method of Example 21 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), to obtain 99.7 mg of yellow solid, with an overall yield of 25.3%. 1 H NMR (400MHz, DMSO-d6) δ13.80(s,1H),10.88(s,1H),10.42(s,1H),7.76(s,1H),7.34(dd,J=8.5,5.6Hz,2H),7.18(t ,J=8.8Hz,2H),7.05(d,J=7.9Hz,1H),6.98(d,J=7.9Hz,1H),6.92(d,J=8.0Hz,1H),5.49(s,2H),3.89(s,2H); EI-MS m / z:393.11[M] - .

[0087] Example 22: Preparation of compound I-22:

[0088] The method of Example 22 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-(trifluoromethyl)benzene was replaced with 1-(azidomethyl)-4-fluorobenzene (5 mmol), to obtain 80.2 mg of yellow solid, with an overall yield of 18.3%. 1H NMR (400MHz, DMSO-d6) δ10.90(s,1H),10.55(s,1H),7.84(s,1H),7.72(d,J=8.3Hz,2H),7.44(d ,J=8.2Hz,2H),7.08-7.01(m,2H),6.91(dd,J=7.3,1.9Hz,1H),5.64(s,2H),3.91(s,2H); EI-MS m / z:443.11[M] - .

[0089] Example 23: Preparation of compound I-23:

[0090] The method of Example 23 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)-1-methylbenzene (5 mmol), to obtain 79.3 mg of yellow solid, with an overall yield of 20.1%. 1 H NMR (400MHz, DMSO-d6) δ12.75(s,1H),10.83(s,1H),10.22(s,1H),7.92(s,1H),7.64(dd,J=8.2,1.1Hz,1H),7.31(t,J=8.5Hz,1H EI-MS m / z:389.4[M] - .

[0091] Example 24: Preparation of compound I-24:

[0092] The method of Example 24 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)-1-methoxybenzene (5 mmol), to obtain 89.1 mg of yellow solid, with an overall yield of 21.9%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H),7.36-7.27(m ,1H),7.26-7.18(m,2H),6.99(dd,J=8.8,1.1Hz,1H),6.86-6.79(m,2H),5.32(t,J=1.0Hz,2H),4.03(s,2H),3.78(s,3H); EI-MS m / z:405.4[M] - .

[0093] Example 25: Preparation of compound I-25:

[0094] The method of Example 25 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with [4-(azidomethyl)phenyl]methaneamine (5 mmol), to obtain 95.4 mg of yellow solid, with an overall yield of 23.5%. 1 H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H ),7.62(dd,J=8.1,1.2Hz,1H),7.40-7.32(m,2H),7.30(d,J=8.5Hz,1H),7.24-7.1 6(m,2H),6.99(dd,J=8.8,1.1Hz,1H),5.31(t,J=1.0Hz,2H),4.03(s,2H),3.95(tt ,J=6.2,1.0Hz,2H),2.76(dt,J=7.1,6.3Hz,1H),2.54(dt,J=7.1,6.3Hz,1H); EI-MS m / z: 404.14 [M] - .

[0095] Example 26: Preparation of compound I-26:

[0096] The method of Example 26 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with 2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)-1-phenylbenzene (5 mmol), to obtain 100.3 mg of yellow solid, with an overall yield of 22.2%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.65-7.55( EI-MS m / z:451.47[M] - .

[0097] Example 27: Preparation of compound I-27:

[0098] The method of Example 27 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)-1-[(trifluoromethyl)oxy]benzene (5 mmol), to obtain 110.3 mg of yellow solid, with an overall yield of 23.9%. 1 H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H),7.36- 7.26(m,2H),7.25(d,J=0.9Hz,1H),7.24-7.16(m,2H),6.99(dd,J=8.8,1.1Hz,1H),5.32(t,J=0.9Hz,2H),4.03(s,2H).

[0099] EI-MS m / z: 459.37 [M] -

[0100] Example 28: Preparation of compound I-28:

[0101] The method of Example 28 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)benzene-1-carboxynitrile (5 mmol), to obtain 99.9 mg of yellow solid, with an overall yield of 24.9%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H), 7.59-7.46(m,4H),7.31(d,J=17.0Hz,1H),6.99(dd,J=8.8,1.1Hz,1H),5.32(t,J=1.0Hz,2H),4.03(s,2H); EI-MS m / z:400.28[M] - .

[0102] Example 29: Preparation of compound I-29:

[0103] The method of Example 29 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)phenol (5 mmol), to obtain 87.5 mg of yellow solid, with an overall yield of 22.3%. 1 H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H EI-MS m / z:391.37[M] - .

[0104] Example 30: Preparation of compound I-30:

[0105] The method of Example 30 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)aniline (5 mmol), to obtain 84.2 mg of yellow solid, with an overall yield of 21.5%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H),7.35(q,J=1.2Hz,1H), 7.34-7.27(m,2H),6.99(dd,J=8.8,1.1Hz,1H),6.56-6.48(m,2H),5.32(t,J=1.0Hz,2H),4.17(d,J=5.7Hz,1H),4.10-4.01(m,3H); EI-MS m / z:390.39[M] - .

[0106] Example 31: Preparation of compound I-31:

[0107] The method of Example 31 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)benzoic acid (5 mmol), to obtain 107.2 mg of yellow solid, with an overall yield of 25.5%. 1 H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),8.12(s,1H),7.92-7.84(m,2H),7.79(s,1H),7.62(dd, EI-MS m / z:419.38[M] - .

[0108] Example 32: Preparation of compound I-32:

[0109] The method of Example 32 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)benzene-1-sulfonamide (5 mmol), to obtain 96.3 mg of yellow solid, with an overall yield of 21.1%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(dd,J=7.0,1.7Hz,3H),7.62(dd,J=8.1,1.2Hz,1H EI-MS m / z:454.45[M] - .

[0110] Example 33: Preparation of compound I-33:

[0111] The method of Example 33 is the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione is replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and [4-(azidomethyl)phenyl](methyl)dioxane-λ is used. 6 Replacing 1-(azidomethyl)-4-fluorobenzene with thion (5 mmol) yielded 98.8 mg of a yellow solid, with an overall yield of 21.8%. 1 H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.79(s,1H),7.76-7.69(m,2H),7.62(dd,J=8.1,1.2Hz,1 EI-MS m / z:453.46[M] - .

[0112] Example 34: Preparation of compound I-34:

[0113] The method of Example 34 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)benzene-1-carboxamide (5 mmol), to obtain 81.9 mg of yellow solid, with an overall yield of 19.5%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),7.92-7.84(m,2H),7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H),7.48-7.40 EI-MS m / z:418.4[M] - .

[0114] Example 35: Preparation of compound I-35:

[0115] The method of Example 35 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with acetic acid-2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 4-(azidomethyl)-N-ethylbenzamide (5 mmol), to obtain 99.4 mg of yellow solid, with an overall yield of 22.2%. 1 H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.89(s,1H),8.01(t,J=4.2Hz,1H),7.86-7.77(m,3H),7.62(dd,J=8.1,1.2Hz,1H),7.46-7 .38(m,2H),7.36-7.27(m,1H),6.99(dd,J=8.8,1.1Hz,1H),5.32(t,J=1.0 Hz, 2H), 4.03 (s, 2H), 3.40 (qd, J=6.3, 4.2Hz, 2H), 1.30-1.22 (m, 3H); EI-MS m / z:446.45[M] - .

[0116] Example 36: Preparation of compound I-36:

[0117] The method of Example 36 was the same as that of Example 19, except that 7-methoxy-1H-indole-2,3-dione was replaced with 2,3-dioxane-2,3-dihydro-1H-indole-7-yl ester (1 mmol), and 1-(azidomethyl)-4-fluorobenzene was replaced with 7-(azidomethyl)quinoline (5 mmol), to obtain 89.5 mg of yellow solid, with an overall yield of 20.9%. 1H NMR(400MHz,Chloroform-d)δ12.61(s,1H),10.00(s,1H),8.91-8.83(m,2H),8.18-8.12(m,1H),8.08(d,J=1.7Hz,1H),7.94(d,J=7.7Hz,1H) ,7.79(s,1H),7.62(dd,J=8.1,1.2Hz,1H),7.54-7.44(m,2H),7.36-7.27(m,1H),6.99(dd,J=8.8,1.1Hz,1H),5.41(s,2H),4.03(s,2H); EI-MS m / z:426.42[M] - .

[0118] Application Examples: Some pharmacological experiments and results of the present invention

[0119] The following are some of the pharmacological experiments and results of the compounds described in this invention:

[0120] The activity of NQO1 inhibitors was determined using the NAD(P)H photoabsorption endpoint method. A 96-well plate was used. 2 μL (1 mM) of the test compound, 2 μL of recombinant human NQO1 protein, and 190 μL of PBS buffer (50 mM, pH 7.4) were added to each well. After incubation at 37°C for 5 min, 4 μL of NADPH (20 mM) was added to each well, and the OD value at 340 nm was immediately detected using a SpectraMax i3x multi-microplate reader. The test lasted for 5 minutes, with measurements taken every 2 seconds. The results were analyzed using a Graphpad Prism 8 to calculate the inhibition rate. The blank control sample contained 192 μL of buffer solution and 2 μL of NQO1 protein, which were incubated before adding 4 μL of NADPH. The positive control sample contained 190 μL of buffer solution, 2 μL of NQO1 protein, and 2 μL of DIC, which were incubated before adding 4 μL of NADPH. The buffer solution contained 0.2 M Na₂HPO₄, 0.2 M NaH₂PO₄, and 0.9% NaCl. The test results for representative compounds are shown in Table 1. Where A represents the IC₂O₁₀ value. 50 Values ​​less than 0.5 μM, B indicates IC50 for NQO1. 50 Values ​​range from 0.5 μM to 1 μM, where C represents the IC50 value for NQO1. 50 Values ​​range from 1 μM to 10 μM, where D represents the IC50 value for NQO1. 50 The value is between 10 μM and 15 μM.

[0121] Table 1. Activity of the compounds in this invention

[0122]

[0123]

[0124] As shown in Table 1, the 2-oxo-1H-quinoline-4-carboxylic acid compounds of the present invention exhibit strong NQO1 enzyme inhibitory activity. Among the compounds in the examples, the compound of Example-22 showed the best NQO1 protein inhibitory activity.

[0125] In addition, seven compounds with inhibitory effects of A were selected and combined with the antitumor drugs LDH inhibitor GNE-140 and gemcitabine to investigate their cytotoxicity against Mia Paca-2 cells. Mia Paca-2 cells were cultured in RPMI-1640 medium (Gibco) containing 10% fetal bovine serum. The inhibitory activity of the compounds against tumor cells was determined by MTT assay. Different tumor cell lines were seeded into 96-well plates and then incubated until cell adhesion. Cells were seeded at a density of 10,000 cells / mL into 96-well microtiter plates and incubated at 37°C for 16 h under a humid atmosphere of 5% CO2. All compounds were prepared as 100 mM stock solutions, and the compounds were diluted to 200 μM with buffer solution, followed by tertiary dilutions to obtain the desired test concentrations. 100 μL of the compound solution and 100 μL of buffer solution were added to each well of the 96-well plate. The 96-well plate was incubated at 37°C for 72 hours. Then, 20 μL of 5 mg / mL MTT solution was added to each well, and the plate was incubated for another 4 hours. After incubation, the solution in each well was aspirated and removed. 150 μL of DMSO solution was added to each well, and the plate was shaken on a shaker for 10 minutes. The OD value at 570 nm was then measured. The test results are shown in Tables 2 and 3.

[0126] Table 2 Results of combined use of some compounds of the present invention with GNE-140

[0127]

[0128]

[0129] As shown in Table 2, the combined drug use experiment results of the representative compounds of the present invention show that the combination of the representative compounds with GNE-140 can enhance its efficacy in killing cancer cells, and the two have a synergistic effect.

[0130] Table 3 Results of combined use of some compounds of the present invention with gemcitabine

[0131]

[0132] As shown in Table 3, the combined drug use experiment results of the representative compounds of the present invention show that the combination of the representative compounds with gemcitabine can enhance the efficacy of killing cancer cells, and the two have a synergistic effect.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound has a chemical structure as shown in (I-a) or (I-b): Among them, R 1 Represents H, -CH3, -OCH3, -F, -OC(O)CH3, or -OH; Ar 1 Selected from , ; In (Ⅰ-a), m is 1, X represents 0, and Ar 2 Selected from , , R 2 Represents H, -Cl, -F, C1-C4 haloalkyl, nitro, -CH3, -CH2NH2, -OCF3, -COOH, -CN, or methoxy; In (Ⅰ-b), m is 0, and X represents CH. 2， Ar 2 for R 2 Represents methylene, R 3 Represents hydrogen, halogen, C1-C4 haloalkyl, C1-C4 alkoxy, C1-C4 alkylamino, cyano, hydroxyl, carboxyl, aminosulfonyl, C1-C4 alkylsulfonyl or C1-C4 amide; Ar 3 Selected from , , .

2. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is selected from any of the following chemical structures: 。 3. A method for preparing the compound according to claim 1, characterized in that, ;or, Among them, R 1 R 2 R 3 Ar 1 Ar 2 and Ar 3 The definition is as described in claim 1.

4. A pharmaceutical composition comprising the compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

5. Use of a compound of claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of an NQO1 inhibitor.

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