Thiazolidinediones compounds containing quinoline structure and their application

Abstract

The present invention relates to the field of pharmaceutical chemistry, and specifically to a class of thiazolidinedione compounds containing quinoline structures and their applications. The compounds discovered by the present invention can affect the interaction between KRAS and the downstream protein cRaf, that is, indirectly inhibit the function of KRAS, thereby inhibiting the proliferation of tumor cells. The compounds and their pharmaceutical preparations can be used to treat a series of diseases caused by abnormal KRAS function, mainly KRAS G12D Related cancers, such as colon cancer, rectal cancer, pancreatic cancer, lung cancer, endometrial cancer, skin cancer, etc.

Description

Technical Field

[0001] The invention relates to the field of medicinal chemistry, and in particular to a class of thiazolidinedione compounds containing a quinoline structure and applications thereof. Background Art

[0002] Quinoline is a multifunctional drug active group, containing a unique skeleton of a variety of natural and synthetic compounds, and has a wide range of biological activities. At the same time, quinoline derivatives also have good anti-tumor activity, such as the treatment of non-small cell lung cancer. Thiazolidinediones containing quinoline structures have also attracted much attention in the medical fields of anti-inflammatory and antibacterial. For example, quinolones have been widely used because of their broad antibacterial activity and good pharmacokinetic properties. Therefore, quinoline, a multifunctional drug active group, has attracted great attention in the field of medicinal chemistry, and its potential application value has made it one of the hot topics in the field of medical research. Quinoline drugs still play an increasingly important role in the field of medical research and development today, especially in the development of anti-tumor drugs. Quinolones show good anti-tumor activity and occupy the main position of biologically active quinoline-containing thiazolidinediones.

[0003] RAS protein is an important protein in the body. After activation, it can transmit signals to downstream cells and bind to cRaf protein to control the proliferation, differentiation and survival of downstream cells. However, high expression or mutation of RAS can cause cancer. RAS includes three subtypes, HRAS, KRAS and NRAS. Among them, KRAS is the main mutation type in tumor cells. KARS mutations mainly include KARS G12C And KARS G12D , currently targeting KARS G12C There have been breakthroughs in the development of inhibitors for KARS, but since KARS lacks a small molecule binding pocket, it is difficult to find small molecule inhibitors with ideal affinity. G12D Discovery of the P110 binding site on the protein for targeting KRAS G12D / KRAS G12D -cRaf provides a new strategy for the development of small molecule drugs.

[0004] Based on the structural characteristics of quinolines and their relationship with KARS G12D Based on the binding mode of the P110 site on the protein, we designed and synthesized a new class of thiazolidinediones containing quinoline structure, which has a strong affinity for KARS G12D It has good selectivity and anti-tumor cell proliferation activity. G12D It provides new ideas for the research and development of anti-tumor drugs. Summary of the invention

[0005] The technical problem solved by the present invention is to provide a class of thiazolidinedione compounds containing quinoline structure or pharmaceutically acceptable salts, isomers, solvates thereof, pharmaceutical compositions thereof and medical uses thereof.

[0006] In order to solve the technical problem of the present invention, the present invention provides the following technical solutions:

[0007] The first aspect of the technical solution of the present invention is to provide a compound as represented by general formula (I) or a pharmaceutically acceptable salt, isomer or solvate thereof.

[0008]

[0009] Formula (I)

[0010] R is:

[0011] , ;

[0012] Preferably, the R is: .

[0013] The pharmaceutically acceptable salt of any of the above compounds of the present invention is trifluoroacetate, hydrochloride and the like.

[0014] The second aspect of the technical solution of the present invention is to provide a method comprising the compound described in the first aspect or a pharmaceutically acceptable salt, isomer, or solvate thereof, and a pharmaceutically acceptable carrier.

[0015] Pharmaceutically acceptable carriers may include any conventional pharmaceutically acceptable carriers, including conventional pharmaceutically acceptable excipients. However, since a pharmaceutically acceptable carrier is not indispensable in the form available for administration, it need not contain excipients that are only relevant to administration. For example, a pharmaceutically acceptable carrier need not contain a lubricant.

[0016] The compounds of the present invention can be added with pharmaceutically acceptable carriers to prepare common pharmaceutical preparations, such as tablets, capsules, powders, syrups, liquids, suspensions, injections, and can be added with common pharmaceutical excipients such as flavors, sweeteners, liquid or solid fillers or diluents.

[0017] The clinical dosage of the compound of the present invention is 0.01-1000 mg / day, and may deviate from this range depending on the severity of the disease or the dosage form.

[0018] The third aspect of the technical solution of the present invention is to provide a method for preparing a method for inhibiting KRAS in cells comprising the compound described in the first aspect or a pharmaceutically acceptable salt, an isomer, a solvate thereof, and the pharmaceutical composition described in the second aspect. G12D Active pharmaceutical application.

[0019] The third aspect of the technical solution of the present invention is to provide a method for preparing a method for treating KRAS comprising the compound described in the first aspect or a pharmaceutically acceptable salt, an isomer, a solvate thereof, and the pharmaceutical composition described in the second aspect. G12D Application in drugs for related cancers.

[0020] The KRAS G12D The relevant cancer is selected from the group consisting of angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma, myxoma, rhabdomyosarcoma, fibroma, lipoma, teratoma, lung cancer, bronchogenic carcinoma, alveolar carcinoma, bronchial adenoma, lymphoma, esophageal cancer, gastric cancer, pancreatic cancer, colon cancer, small intestine cancer, large intestine cancer, rectal cancer, kidney cancer, lymphoma, leukemia, bladder and urethra cancer, prostate cancer, testicular cancer, liver cancer, bile duct cancer, hepatoblastoma, hepatocellular adenoma, gallbladder cancer, bile duct cancer, bone cancer, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, malignant lymphoma, multiple myeloma, malignant giant cell carcinoma Cell tumors Chordoma, osteochondroma, benign chondroma, chondroblastoma, chondromyxoid fibroma, glioma, germ cell tumor, glioblastoma multiforme, oligodendroglioma, neurothecoma, retinoblastoma, congenital tumors, spinal neurofibroma, endometrial carcinoma, malignant teratoma, vulvar squamous cell carcinoma, fallopian tube carcinoma, myeloid leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, multiple myeloma, malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi sarcoma, lipoma, hemangioma, cutaneous fibroma.

[0021] Preferably, the cancer is lung cancer, colon cancer, rectal cancer or pancreatic cancer.

[0022] More preferably, the lung cancer is non-small cell lung cancer or small cell lung cancer.

[0023] Beneficial technical effects:

[0024] The advantage of the present invention is that KRAS and its mutant proteins have smooth surfaces and small active cavities, and it is difficult to find compounds with ideal affinity. The compounds found in the present invention can affect the interaction between KRAS and the downstream protein cRaf, that is, indirectly inhibit the function of KRAS, and then inhibit the proliferation of tumor cells. Such compounds and their pharmaceutical preparations can be used to treat a series of diseases caused by abnormal KRAS function, mainly KRAS G12D Related cancers, such as colon cancer, rectal cancer, pancreatic cancer, lung cancer, endometrial cancer, skin cancer, etc. DETAILED DESCRIPTION

[0025] The present invention will be further described below in conjunction with specific implementation modes so that those skilled in the art can better understand the present invention, but the present invention is not limited thereto.

[0026] The compounds of the general formula (I) of the present invention can be prepared by the following method:

[0027]

[0028] Where R is:

[0029]

[0030] raw material N -Boc-bromoethylamine (3) and 2,4-thiazolidinedione (4) undergo nucleophilic substitution reaction under the action of catalyst TBAI to obtain thiazolidinedione compound W7, compound W3 undergoes substitution reaction with electrophilic reagent to obtain quinoline ester compound W8, compound W8 is reduced to aldehyde group under the reduction of reducing agent DIBAL-H to obtain quinoline aldehyde compound W9, compound W9 undergoes aldol condensation reaction with intermediate compound W7 to obtain thiazolidinedione compound W10 containing quinoline structure, and finally compound W10 removes Boc group under acidic conditions to obtain target compound CR3.

[0031] Example 1 Synthesis of (Z)-3-(2-aminoethyl)-5-((4-(benzyl)quinolinyl)methylene)thiazolidine-2,4-dione trifluoroacetate (C44):

[0032]

[0033] The raw material N-Boc-bromoethylamine (3) (4.6 g, 20.51 mmol) and 2,4-thiazolidinedione (4) (2.0 g, 17.09 mmol) were dissolved in acetonitrile (100 mL), and tetrabutylammonium iodide (3.41 mmol) and potassium carbonate (3.3 g, 20.92 mmol) were added to react at room temperature for 12 h. The reaction solution was evaporated under reduced pressure to remove the solvent, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and evaporated under reduced pressure to remove the solvent. The crude product was separated and purified by silica gel column chromatography (mobile phase: n-hexane-ethyl acetate = 5:1) to obtain the intermediate compound W7. 4-Oxo-1,4-dihydroquinoline-2-carboxylic acid methyl ester W3 (1 mmol) was dissolved in acetonitrile (2 mL), and anhydrous potassium carbonate was slowly added. Under nitrogen protection, the electrophilic reagent benzyl bromide was slowly added, and the reaction was refluxed for 12 h. The reaction solution was quenched with water, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The crude product was separated by silica gel column chromatography (mobile phase: dichloromethane: methanol = 30:1) to obtain compound W8. Compound W8 (1.0 g, 3.41 mmol) was added to the reaction flask. After nitrogen replaced the air, anhydrous dichloromethane (50 mL) was added to the reaction system and heated at -80 °C. oC for 30 min, DIBAL-H (1 mL, 4.09 mmoL) was added dropwise to the reaction system and reacted for 12 h. The reaction solution was quenched with 1N hydrochloric acid, extracted with dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The crude product was separated by silica gel column chromatography (mobile phase: dichloromethane: methanol = 30:1) to purify the quinoline aldehyde compound W9. Compound W9 (218 mg, 0.85 mmoL) and intermediate W7 (280 mg, 1.07 mmoL) were dissolved in anhydrous ethanol (40 mL), and after nitrogen replaced the air, piperidine (120 μL) was added to the reaction system and reacted for 8 h. The raw material disappeared when detected by TLC (developing solvent: dichloromethane-methanol = 30:1). The solvent was removed by rotary evaporation under reduced pressure, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by evaporation under reduced pressure. The crude product was purified by silica gel column chromatography (mobile phase: dichloromethane: methanol = 100:1) to obtain the thiazolidinedione compound W10 containing a quinoline structure. W10 (100 mg, 0.25 mmol) was dissolved in anhydrous dichloromethane (10 mL). After nitrogen replaced the air, trifluoroacetic acid (40 μL) was added to the reaction system and reacted for 8 h. The solvent was removed by evaporation under reduced pressure, and the crude product was purified by silica gel column chromatography (mobile phase: dichloromethane: methanol = 90:1) to obtain the target compound C44. White needle-like solid, 53%, mp: 148-150 o C. 1 H NMR (800 MHz, DMSO- d 6) δ 8.18 (d, J = 8.2 Hz, 1H), 8.08 (d, J = 8.3 Hz, 1H), 8.04 (s,1H), 7.84 (t, J = 7.5 Hz, 1H), 7.75 (s, 1H), 7.65 – 7.62 (m, 1H), 7.61 (d, J =7.4 Hz, 2H), 7.47 (t, J = 7.4 Hz, 2H), 7.41 (t, J = 7.2 Hz, 1H), 5.45 (s, 2H), 3.93 (t, J = 5.6 Hz, 2H), 3.12 (s, 2H). IR (KBr, cm -1): 721.41, 734.92, 761.90,798.40, 836.45, 1070.40, 1120.83, 1155.56, 1177.52, 1199.43, 1238.63,1342.67, 1361.71, 1397.99, 1425.97, 1454.53, 1505.07, 1519.13, 1580.33,1625.10, 1686.52, 1738.47, 3049.28. HRMS (ESI): m / z Calculate for C 22 H 19 N3O3S[M+H]+:406.1225 found: 406.1210.

[0034] The preparation method of compound C46 in Example 2 is the same as that in Example 1, except that different electrophilic reagents are used. The electrophilic reagents used in the specific examples are described in the corresponding examples:

[0035] Example 2 Synthesis of (Z)-3-(2-aminoethyl)-5-((4-((3,5-bis(trifluoromethyl)benzyl)oxy)quinolin-2-yl)methylene)thiazolidine-2,4-dione trifluoroacetate (C46):

[0036]

[0037] The electrophilic reagent used is 3,5-bis(trifluoromethyl)benzyl chloride. White needle-like solid, 53%, mp: 156-158 o C. 1 HNMR (800 MHz, DMSO- d 6) δ 8.34 (s, 2H), 8.19 (d, J = 8.8 Hz, 2H), 8.10 (d, J = 8.4Hz, 1H), 8.06 (s, 1H), 7.87 – 7.83 (m, 1H), 7.77 (s, 1H), 7.66 (t, J = 7.5 Hz,1H), 5.63 (s, 2H), 3.94 (t, J = 5.9 Hz, 2H), 3.12 (q, J = 5.4 Hz, 2H). IR (KBr,cm -1): 437.97, 448.26, 502.64, 512.64, 682.66, 702.26, 722.58, 743.21, 768.04,798.22, 843.86, 881.41, 1124.99, 1146.59, 1170.60, 1279.98, 1357.11, 1391.12,1427.52, 1464.47, 1505.86, 1585.16, 1624.44, 1682.99, 1738.13, 3045.90. HRMS(ESI): m / z Calculate for C 24 H 17 F6N3O3S[M+H]+: 542.0973 found: 542.0958.

[0038] Some biological activity tests and results of the compounds of the present invention are as follows:

[0039] Experimental Example 1: Effects of the compounds of the present invention on KRAS G12D -cRaf affinity evaluation

[0040] 1.1. Experimental methods:

[0041] 0.1 μL of compound solution was transferred to a 384-well plate. The initial concentration of the test compound was 1 μM, and the test compound was diluted 3-fold to 10 concentrations. 2.5 μL of KRAS was added to each well. G12D , centrifuged at 1000 rpm for 1 min, added 2.5 μL cRAF to each well, centrifuged at 1000 rpm for 1 min, and incubated at 25°C for 15 min. Add 5 μL SA-Tb&GST-XL665 to the 384-well plate, centrifuged at 1000 rpm for 1 min, and incubated at 25°C for 60 min. HTRF signals (665 / 620 nm) were read using a BMG microplate reader. Inhibition rate or IC was calculated using GraphPadPrism software 50 value.

[0042] 1.2. Experimental results

[0043] 1. Single concentration inhibition rate results

[0044] Table 1-1 Effects of target compounds on KRAS G12D -cRaf affinity (10 μM)

[0045]

[0046] Preliminary screening results showed that at a concentration of 10 µM, CR series compounds C44 and C46 had significant effects on KRAS G12D -cRaf has a certain inhibitory effect.

[0047] 2. CR series compounds for KRAS G12D -cRaf IC 50 value

[0048] Table 1-2 Effects of CR series compounds on KRAS G12D -cRaf IC 50 value

[0049]

[0050] The compounds with better single concentration inhibitory effects were further tested for KRAS G12D -cRaf IC 50 The results showed that compound C46 had an G12D -cRaf has a strong inhibitory effect.

[0051] Experimental Example 2: Determination of the inhibitory effect of target compounds on tumor cell growth

[0052] 2.1 Experimental Materials

[0053] The compound of the present invention is dissolved in dimethyl sulfoxide and prepared at 1 mg / mL in RPMI-1640 medium containing 10% fetal bovine serum for later use. When group administration is performed, the compound is diluted with the medium at a certain multiple to the required concentration.

[0054] Table 2-1 Experimental reagents

[0055]

[0056] MTT assay method

[0057] NCIH-1385 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at the following conditions: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0058] PANC-1 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0059] AsPc-1 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0060] LOVO cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and passaged every 2-3 days. The cell culture incubator was set at: CO2 concentration: 5%, temperature: 37 o C, Cells in the logarithmic growth phase were used for the experiment.

[0061] NCIH-1385 cells, PANC-1 cells, AsPc-1 cells, LOVO cells (8 × 10 3 After 24 hours of incubation, the substance to be measured was added and incubated for 48 hours. After the cell supernatant was removed, 100 μL of MTT aqueous solution with a final concentration of 0.5 mg / mL was added to each well, and the culture plate was placed in an incubator to continue incubating for 4 hours. After the cell supernatant was removed, 100 μL of DMSO was added, and the culture plate was shaken and the absorbance (OD) at 490 nm was measured with an enzyme marker. Cell activity % = OD value of the drug group / OD value of the blank group * 100%.

[0062] 2.3 Measurement results

[0063] The synthesized compounds can inhibit the proliferation of tumor cells to a certain extent, which is comparable to the positive drug cisplatin, especially compound C46, ​​which has an IC 50 Better than the positive drug cisplatin (Table 2-2).

[0064] Table 2-2 IC of preferred compounds against tumor cell proliferation 50

[0065]

[0066] Note: CDDP is the positive control drug cisplatin.

[0067] 2.4 Discussion

[0068] The thiazolidinedione compounds containing quinoline structure of the present invention have certain growth inhibition on tumor cells, among which C46 shows good anti-proliferation activity in four types of tumor cells. In particular, C46 not only shows the same anti-tumor cell proliferation activity as the positive drug cisplatin in LOVO cells, but also has a significant effect on KRAS G12DHighly expressed AsPc-1 cells showed better anti-tumor cell proliferation activity than the positive drug cisplatin, laying the foundation for the development of anti-cancer drugs.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The structural formula of the compound is: 、 。 2. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 2 in the preparation of a drug for inhibiting KRAS in cells G12D Active pharmaceutical application.

4. The compound according to claim 1 or its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 2 in the preparation of a pharmaceutical composition for treating KRAS G12D Application in drugs for related cancers.

5. The use according to claim 4, characterized in that: The cancer is lung cancer, colon cancer, rectal cancer or pancreatic cancer.

6. The use according to claim 5, characterized in that: The lung cancer is non-small cell lung cancer.

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