N-phenylbenzenesulfonamide compound, and preparation method and application thereof

Abstract

The invention discloses an N-phenylbenzenesulfonamide compound as well as a preparation method and application thereof. The compound disclosed by the invention is shown as a formula (I). The compound is an LDHA enzyme inhibitor, is a sulfonamide compound with anti-tumor activity, shows good anti-tumor activity at the cellular level, has good safety and can be used for preparing anti-tumor drugs. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to an organic compound, a preparation method and application thereof, and in particular to an N-phenylbenzenesulfonamide compound, a preparation method and application thereof. Background Art

[0002] LDHA enzyme, or lactate dehydrogenase, is a type of oxidoreductase widely found in mammalian cells. It reduces pyruvate to lactate with the assistance of reduced nicotinamide adenine dinucleotide NADH cofactor. There are five subtypes of LDH enzymes, of which LDHA has a higher affinity for pyruvate and performs anaerobic metabolism and pyruvate reduction, while LDHB tends to participate in its reverse reaction.

[0003] Under normal circumstances, the body's sugar metabolism tends to be from glucose metabolized to pyruvate, which then enters the mitochondria and undergoes aerobic metabolism of oxidative phosphorylation to generate energy. Since the abnormal growth and reproduction of tumor cells require sufficient and rapid energy supply and biosynthetic intermediates, tumor cells switch from oxidative phosphorylation to aerobic glycolysis. This special phenomenon caused by metabolic reprogramming provides the basis for LDH enzyme to become a focus of cancer treatment. Since it plays a key role in the glycolysis process of tumors, the inhibition of LDH enzyme can target cancer cell glycolysis, selectively starve cancer cells, and thus inhibit their growth and reproduction activity. It is worth studying in the field of cancer treatment research targeting cancer cell metabolic reprogramming.

[0004] Currently, chemotherapy regimens such as gemcitabine, fluorouracil, and S-1 are commonly used to treat advanced pancreatic cancer. When using these anti-tumor drugs, patients often experience adverse reactions such as bone marrow suppression, gastrointestinal side effects, lung toxicity, and organ damage. Summary of the invention

[0005] Purpose of the invention: The present invention provides N-phenylbenzenesulfonamide compounds, which can be used as inhibitors of LDH enzymes to treat pancreatic cancer, and at the same time exhibit good anti-tumor effects at the cellular level, and have good safety and reduced side effects. Another purpose of the present invention is to provide a preparation method and use of the N-phenylbenzenesulfonamide compounds, which can be used to prepare anti-tumor drugs.

[0006] Technical solution: The N-phenylbenzenesulfonamide compound or its pharmaceutically acceptable salt described in this solution has a structure as shown in the general formula (I):

[0007]

[0008] Wherein P1 represents phenyl, 3-7 membered cycloalkyl, 3-7 membered heterocycloalkyl; the 3-7 membered heterocycloalkyl is a 3-7 membered ring containing two or three elements of C, N, and O;

[0009] X represents -(CH2) n -NH-, amide or absent, n is selected from 0, 1, 2, 3, 4;

[0010] R1 represents hydrogen or C1-C6 alkyl;

[0011] R2 represents hydrogen, monosubstituted or polysubstituted halogen, cyano, C 1～4 One or more of alkoxy and trifluoromethyl;

[0012] R3 represents hydrogen, C1-C6 alkyl, acetyl, trifluoroacetyl, oxalyl, tert-butyloxycarbonyl, or C1-C4 chain-linked carboxyl;

[0013] R4 represents hydrogen, monosubstituted or polysubstituted halogen, cyano, C 1～4 Alkoxy.

[0014] In the N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof, P1 represents a phenyl group, a 3-6-membered cycloalkyl group, or a 3-6-membered heterocycloalkyl group; the 3-6-membered heterocycloalkyl group is a 3-6-membered ring containing two or three elements of C, N, and O.

[0015] In the N-phenylbenzenesulfonamide compound or its pharmaceutically acceptable salt, P1 represents a phenyl group, a 3- to 6-membered cycloalkyl group, a piperazinyl group, or a morpholinyl group.

[0016] The N-phenylbenzenesulfonamide compound or its pharmaceutically acceptable salt, R2 represents hydrogen, monosubstituted or polysubstituted halogen, cyano, C 1～4 One, two or three of alkoxy and trifluoromethyl.

[0017] The N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof is selected from the following compounds:

[0018]

[0019]

[0020]

[0021]

[0022] The preparation method of the N-phenylbenzenesulfonamide compound or its pharmaceutically acceptable salt,

[0023] When X is absent and P1 is a nitrogen-containing aliphatic ring, compound IV and The target compound I is obtained by reaction at a reaction temperature of 80 to 130° C., wherein R1, R2, R3, R4, and P1 are defined as above. R3 is substituted piperazinyl or morpholinyl:

[0024]

[0025] When X is -(CH2) n -NH-, n is selected from 0, 1, 2, 3, 4, compound IV and The target compound II is obtained by reaction at a reaction temperature of 80 to 130° C., wherein R1, R2, R3, R4, and P1 are defined as above. -(CH2) substituted by R3 n -phenyl, 3-7 membered cycloalkyl-(CH2) n -; n is selected from 0, 1, 2, 3, 4:

[0026]

[0027] When X is an amide group, compound V and The target compound III is obtained by reaction at room temperature, wherein R1, R2, R3, R4, and P1 are defined as above. P1 substituted with R3 and COCl:

[0028]

[0029] The pharmaceutical composition contains the N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0030] The use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of an LDH enzyme inhibitor. Preferably, the use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of an LDHA enzyme inhibitor.

[0031] The compound or a pharmaceutically acceptable salt thereof is used in preparing a drug for treating malignant tumors.

[0032] The use, the malignant tumor is pancreatic cancer.

[0033] Preferably, in the N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof, the halogen is selected from fluorine, chlorine, bromine and iodine.

[0034] The method for preparing the N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof of the present invention, when X is absent and P1 is a nitrogen-containing aliphatic ring, compound IV and The target compound I is obtained by reaction, the reaction temperature is 110°C, the reaction time is 8-12h, and the reaction solvent can be 1,4-dioxane, tetrahydrofuran, DMF, etc. An organic base or an inorganic base, such as triethylamine, sodium tert-butoxide, potassium carbonate, BINAP, etc., is added to the reaction. A palladium catalytic reagent, such as tridibenzylideneacetone dipalladium, bis(3,5,3′,5′-dimethoxydibenzylideneacetone)palladium(0), bis(dibenzylideneacetone)palladium, etc., is also added to the reaction; at the same time, the target compound I can be purified by common separation methods, such as recrystallization, column chromatography, etc.:

[0035]

[0036] The present invention also includes hydrates, stereoisomers, solvates and pharmaceutically acceptable salts of the compounds of general formula I. They have the same pharmacological activity as the compounds of general formula I.

[0037] 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.

[0038] Beneficial effects: Compared with the prior art, the present invention has the following advantages: the compound of the present invention or its pharmaceutically acceptable salt is an LDH inhibitor, can promote tumor cell apoptosis, can be used to treat pancreatic cancer, and has good application prospects. DETAILED DESCRIPTION

[0039] The following examples are provided to illustrate the present invention in detail. In the present invention, the following examples are provided to better illustrate the present invention and are not intended to limit the scope of the present invention.

[0040] Example 1: Preparation of Compound I-1

[0041] 3-Bromobenzenesulfonyl chloride (0.5 g) and 2-chloroaniline (0.25 g) were dissolved in anhydrous DCM (50 mL), followed by the addition of pyridine (0.31 g). The reaction solution was stirred at room temperature overnight. After the reaction was completed, DCM was used for extraction, and the reaction solution was adjusted to neutrality by adding dilute HCl. The organic phase was washed with saturated NaCl solution (30 mL) and dried over anhydrous Na2SO4. Purification was performed by silica gel column chromatography using petroleum ether / ethyl acetate (10:1) as the mobile phase to obtain white solid N-(2-chloro-5-cyanophenyl)-3-bromobenzenesulfonamide (400.49 mg, 59.1%). N-(2-chloro-5-cyanophenyl)-3-bromobenzenesulfonamide (400.5 mg), methylpiperazine (0.58 g), bis(3,5,3′,5′-dimethoxydibenzylideneacetone)palladium(0) (93.7 mg), sodium tert-butoxide (1.11 g), 2-(dicyclohexylphosphine)-3,6-dimethoxy-2′-4′-6′-tri-1-propyl-11′-biphenyl (BrettPhos, 61.7 mg) were dissolved in 1,4-dioxane (50 mL). The reaction solution was stirred at 110° C. for 10 h. After the reaction was completed, the mixture was cooled to room temperature, the solvent was dried by rotary evaporation, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated NaCl, dried over anhydrous Na2SO4, and purified by silica gel column chromatography with dichloromethane / methanol (60:1) to obtain a colorless oily compound I-1 (78.18 mg, 18.5%). 1 H NMR (400MHz, DMSO-d6) δ7.91 (s, 1H), 7.83 (d, J = 1.7Hz, 1H), 7.46 (d, J = 8.2Hz, 1H), 7.36-7.33 (m, 2H), 7.21 (d, J = 2.8Hz, 1H), 7.0 9 (d, J=2.1Hz, 1H), 7.04 (s, 1H), 3.60-3.54 (m, 2H), 3.20-3.13 (m, 2H), 2.60 (t, J=5.1Hz, 2H), 2.46 (t, J=5.1Hz, 2H), 1.99 (s, 3H).

[0042] Example 2: Preparation of Compound I-2

[0043] Using 2,5-dichloroaniline (0.3 g) instead of 2-chloroaniline, the compound I-2 (79.9 mg, 17.3%) was synthesized in the same manner as compound I-1. 1H NMR (400MHz, Chloroform-d) δ10.39 (s, 1H), 7.46 (ddd, J=11.4, 4.5, 2.4Hz, 1H), 7.37 (dt, J=7.5, 3.8Hz, 1H), 7.26 (dd, J=7. 5, 1.9Hz, 2H), 7.23 (d, J=3.0Hz, 1H), 6.98 (ddt, J=10.6, 5.4, 2.6Hz, 2H), 3.61-3.36 (m, 4H), 2.69-2.48 (m, 4H), 2.07 (s, 3H).

[0044] Example 3: Preparation of Compound I-3

[0045] Using 2-chloro-5-fluoroaniline (0.29 g) instead of 2-chloroaniline, the compound I-3 (69.5 mg, 15.6%) was synthesized in the same manner as compound I-1. 1 H NMR (400MHz, Chloroform-d) δ10.10 (s, 1H), 7.26-7.18 (m, 2H), 7.13 (d, J=8.7Hz, 1H), 7.04 (s, 1H), 7.02 (s, 1H), 6. 68 (d, J=8.7Hz, 1H), 6.57 (dd, J=15.2, 7.9Hz, 1H), 3.30 (dd, J=7.4, 5.7Hz, 4H), 3.12 (q, J=7.3Hz, 4H), 1.99 (s, 3H).

[0046] Example 4: Preparation of Compound I-4

[0047] Using 2-chloro-5-methoxyaniline (0.31 g) instead of 2-chloroaniline, the compound I-4 (74.9 mg, 16.3%) was synthesized in the same manner as compound I-1. 1 H NMR (300MHz, Chloroform-d) δ10.20 (s, 1H), 7.55 (d, J=1.7Hz, 1H), 7.54-7.50 (m, 3H), 7.49 (d, J=3.7Hz, 1H), 7. 39 (d, J=1.7Hz, 1H), 7.35 (t, J=1.4Hz, 1H), 3.73 (s, 3H), 3.15 (qd, J=7.3, 4.7Hz, 4H), 2.32 (m, 4H), 1.99 (s, 3H).

[0048] Example 5: Preparation of Compound I-5

[0049] Using 2-chloro-5-trifluoromethylaniline (0.38 g) instead of 2-chloroaniline, the compound I-5 (80.0 mg, 15.9%) was synthesized in the same manner as compound I-1. 1 H NMR (400MHz, Chloroform-d) δ9.94 (s, 1H), 7.32 (d, J=8.6Hz, 2H), 7.27-7.24 (m, 1H), 7.20 (t, J=7.9Hz, 2H), 7.05 (t d, J=7.5, 1.4Hz, 1H), 6.98 (td, J=7.7, 1.7Hz, 1H), 3.30 (qd, J=7.3, 5.5Hz, 4H), 3.00 (q, J=7.3Hz, 4H), 1.99 (s, 3H).

[0050] Example 6: Preparation of Compound I-6

[0051] Using 2-chloro-5-cyanoaniline (0.30 g) instead of 2-chloroaniline, the compound I-6 (76.6 mg, 16.9%) was synthesized in the same manner as compound I-1. 1 H NMR (300MHz, Chloroform-d) δ10.21 (s, 1H), 7.42 (s, 1H), 7.31 (d, J=1.5Hz, 1H), 7.25 (dd, J=4.6, 2.2 Hz, 3H), 7.14 (d, J=8.9Hz, 1H), 7.08-7.02 (m, 1H), 3.38-3.19 (m, 4H), 2.79-2.62 (m, 4H), 2.41 (s, 3H).

[0052] Example 7: Preparation of Compound I-7

[0053] Compound I-7 (76.3 mg, 15.7%) was synthesized in the same manner as compound I-1, except that 2-chloro-5-cyanoaniline (0.30 g) was used instead of 2-chloroaniline and 1-acetylpiperazine (0.74 g) was used instead of methylpiperazine. 1H NMR (400MHz, Chloroform-d) δ7.94 (d, J=1.9Hz, 1H), 7.43 (d, J=8.3Hz, 1H), 7.38 (q, J=2.3, 1.7Hz, 1H), 7.34 (d, J=1.9Hz, 1H), 7.33 (dd, J=3.0, 1.7Hz, 1H ), 7.30-7.29 (m, 1H), 7.25 (s, 1H), 7.11 (dd, J=8.5, 2.5Hz, 1H), 3.79 (t, J=5 .3Hz, 2H), 3.66 (t, J=5.1Hz, 2H), 3.23 (dt, J=16.8, 5.3Hz, 4H), 2.17 (s, 3H).

[0054] Example 8: Preparation of Compound I-8

[0055] 2-Chloro-5-cyanoaniline (0.30 g) was used to replace 2-chloroaniline, and 1-Boc-piperazine (1.07 g) was used to replace methylpiperazine. The same method as compound I-1 was used to synthesize tert-butyl 4-(3-(N-(2-chloro-5-cyanophenyl)sulfonamido)phenyl)piperazine-1-carboxylate (88.53 mg, 16.0%). Tert-butyl 4-(3-(N-(2-chloro-5-cyanophenyl)sulfonamido)phenyl)piperazine-1-carboxylate (88.5 mg) was dissolved in HCl·EA (10 mL), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, N-(2-chloro-5-cyanophenyl)-3-piperazinebenzenesulfonamide (71.0 mg, 97.8%) was obtained by filtration. N-(2-chloro-5-cyanophenyl)-3-piperazine benzenesulfonamide (70 mg) was dissolved in acetonitrile (30 mL), methyl bromoacetate (29.1 mg) and TEA (38.5 mg) were added, and the reaction solution was stirred at room temperature for 4 h. After the reaction was completed, ethyl acetate was used for extraction, and the organic phase was washed with saturated NaCl and dried over anhydrous Na2SO4 to obtain methyl 2-(4-(3-(N-(2-chloro-5-cyanophenyl)sulfonamido)phenyl)piperazine)acetate (44.4 mg, 52.0%). Methyl 2-(4-(3-(N-(2-chloro-5-cyanophenyl)sulfonamido)phenyl)piperazine)acetate (44.4 mg) was dissolved in THF (10 mL), and then LiOH aqueous solution (5 mL) was added, and the reaction solution was heated to 80°C and stirred for 4 h. After the reaction was completed, ethyl acetate was used for extraction, the pH was adjusted to acidic, the organic phase was washed with saturated NaCl, and dried over anhydrous Na2SO4 to obtain compound I-8 (38.3 mg, 89.1%). 1H NMR (400MHz, Chloroform-d) δ8.68 (t, J=2.0Hz, 1H), 8.48 (ddd, J=8.3, 2.2, 1.1Hz, 1H), 8.13 (dt, J=7.9, 1.3Hz, 1H), 8 .00 (d, J=1.7Hz, 1H), 7.75 (t, J=8.1Hz, 1H), 7.52-7.38 (m, 2H), 3.65 (s, 2H), 2.45-2.20 (m, 4H), 2.03 (d, J=6.9Hz, 4H).

[0056] Example 9: Preparation of Compound I-9

[0057] 2-Chloro-5-cyanoaniline (0.30 g) was used to replace 2-chloroaniline, and 1-Boc-piperazine (1.07 g) was used to replace methylpiperazine. The same method as compound I-1 was used to synthesize tert-butyl 4-(3-(N-(2-chloro-5-cyanophenyl)sulfonylamino)phenyl)piperazine-1-carboxylate (88.53 mg, 16.0%). Tert-butyl 4-(3-(N-(2-chloro-5-cyanophenyl)sulfonylamino)phenyl)piperazine-1-carboxylate (88.5 mg) was dissolved in HCl·EA (10 mL), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, N-(2-chloro-5-cyanophenyl)-3-piperazinebenzenesulfonamide was obtained by filtration.

[0058] (71.0 mg, 97.8%). N-(2-chloro-5-cyanophenyl)-3-piperazinebenzenesulfonamide (70 mg) was dissolved in DCM (30 mL), trifluoroacetic anhydride (43.9 mg) and TEA (38.5 mg) were added, and the reaction solution was heated to 40°C and stirred for 5 h. After the reaction was completed, ethyl acetate was used for extraction, and the organic phase was washed with saturated NaCl and dried over anhydrous Na2SO4 to obtain compound I-9 (37.0 mg, 41.2%). 1 H NMR (400MHz, DMSO-d6) δ9.33-9.21 (m, 1H), 7.40 (d, J=8.2Hz, 1H), 7.21 (d, J=8.1Hz, 1H), 7.17 (dd, J=5.2, 3.0 Hz, 1H), 7.12 (d, J=2.0Hz, 1H), 7.00 (ddd, J=7.9, 5.1, 2.1Hz, 2H), 6.93 (dd, J=8.2, 2.0Hz, 1H), 3.48-3.31 (m, 4H), 3.18 (t, J=4.9Hz, 4H).

[0059] Example 10: Preparation of Compound I-10

[0060] Compound I-10 (38.2 mg, 8.81%) was synthesized in the same manner as compound I-8, except that methyl bromopropionate (31.7 mg) was used instead of methyl bromoacetate. 1 H NMR (300MHz, Chloroform-d) δ7.37-7.28 (m, 4H), 6.95 (dd, J=1 1.7, 7.8Hz, 3H), 3.86-3.76 (m, 2H), 3.71-3.61 (m, 2H), 3.47 (d, J=25.4Hz, 2H), 3.20 (dt, J=10.3, 5.2Hz, 4H), 2.52-2.44 (m, 2H).

[0061] Example 11: Preparation of Compound I-11

[0062] Compound I-11 was synthesized in the same manner as compound I-1, using 2-chloro-5-cyanoaniline (0.30 g) instead of 2-chloroaniline and 1-Boc-piperazine (1.07 g) instead of methylpiperazine. 1 H NMR (400MHz, Chloroform-d) δ7.95 (d, J=1.9Hz, 1H), 7.42 (d, J=8.3Hz, 1H), 7.39 (d, J=8.0Hz, 1H), 7.35 (dd, J=5.3, 3.0 Hz, 2H), 7.32 (d, J=1.9Hz, 1H), 7.30 (d, J=1.6Hz, 1H), 7.21-7.13 (m, 2H), 3.62 (t, J=5.2Hz, 4H), 3.20 (t, J=5.2Hz, 4H), 1.51 (s, 9H).

[0063] Example 12: Preparation of Compound I-12

[0064] 2-Chloro-5-cyanoaniline (0.30 g) was used to replace 2-chloroaniline, and 1-Boc-piperazine (1.07 g) was used to replace methylpiperazine. Compound I-11 (88.53 mg, 16.0%) was synthesized in the same manner as compound I-1. Compound I-11 (88.5 mg) was dissolved in HCl·EA (10 mL), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, the compound I-12 (71.0 mg, 97.8%) was obtained by filtration. 1H NMR (300MHz, DMSO-d6) δ10.45 (s, 1H), 9.44 (s, 1H), 7.76-7.64 (m, 2H), 7.62 (d, J=1.7Hz, 1H), 7.50-7.3 8 (m, 1H), 7.29 (dt, J=3.9, 2.3Hz, 2H), 7.19 (d, J=7.6Hz, 1H), 4.97 (d, J=32.3Hz, 4H), 3.44-3.36 (m, 4H).

[0065] Example 13: Preparation of Compound I-13

[0066] Compound I-13 (69.2 mg, 15.8%) was synthesized in the same manner as compound I-1, except that 2-chloro-5-cyanoaniline (0.30 g) was used instead of 2-chloroaniline and morpholine (0.50 g) was used instead of methylpiperazine. 1 H NMR (300MHz, Chloroform-d) δ7.96 (d, J=1.8Hz, 1H), 7.44 (s, 1H), 7.41 (s, 1H), 7.38 (s, 1H), 7.36-7.3 4 (m, 1H), 7.31 (d, J=1.3Hz, 1H), 7.16 (s, 1H), 7.12-7.08 (m, 1H), 3.96-3.83 (m, 4H), 3.30-3.14 (m, 4H).

[0067] Example 14: Preparation of Compound I-14

[0068] Use 2-chloro-5-cyanoaniline (0.30g) instead of 2-chloroaniline, and use 3-phenylbenzenesulfonyl chloride (0.49g) instead of 3-bromobenzenesulfonyl chloride, and synthesize compound I-14 (390.3mg, 53.9%) by the same method as compound I-1 to synthesize N-(2-chloro-5-cyanophenyl)-3-bromobenzenesulfonamide. 1 H NMR (400MHz, Chloroform-d) δ8.06 (t, J=1.9Hz, 1H), 8.02 (d, J=1.9Hz, 1H), 7.82 (dddd, J=14.2, 7.9, 1.9, 1.1Hz, 2H), 7.59 (d, J =7.8Hz, 1H), 7.57-7.52 (m, 2H), 7.50 (ddd, J=7.7, 6.7, 1.4Hz, 2H), 7.46-7.40 (m, 2H), 7.35 (dd, J=8.3, 1.9Hz, 1H), 7.23 (s, 1H).

[0069] Example 15: Preparation of Compound I-15

[0070] Compound I-15 (37.3 mg, 8.54%) was synthesized in the same manner as compound I-8, using monomethyl oxalyl chloride (23.1 mg) instead of methyl bromoacetate. 1 H NMR (400MHz, DMSO-d6) δ9.39 (s, 1H), 7.40 (d, J = 8.2Hz, 1H), 7.22-7.15 (m, 2H), 7.13 (d, J = 2.0Hz, 1H), 6 .99 (ddd, J=7.8, 4.8, 2.1Hz, 2H), 6.93 (dd, J=8.1, 2.0Hz, 1H), 3.50-3.31 (m, 4H), 3.18 (t, J=5.2Hz, 4H).

[0071] Example 16: Preparation of Compound I-16

[0072] 3-nitrobenzenesulfonyl chloride (0.43 g) was used instead of 3-bromobenzenesulfonyl chloride and 2-chloro-5-cyanoaniline was used.

[0073] (0.30g) was used to replace 2-chloroaniline, and N-(2-chloro-5-cyanophenyl)-3-nitrobenzenesulfonamide (337.6mg, 51.1%) was synthesized in the same manner as compound I-1. N-(2-chloro-5-cyanophenyl)-3-nitrobenzenesulfonyl (337.6mg), reduced iron powder (279.3mg), and ammonium chloride (267.5mg) were dissolved in a solvent of ethanol: water = 3:1. The reaction solution was stirred at 80°C for 4h. After the reaction was completed, it was cooled to room temperature, filtered with diatomaceous earth, the solvent was dried by spin drying, and extracted with ethyl acetate. The organic phase was washed with saturated NaCl and dried over anhydrous Na2SO4 to obtain N-(2-chloro-5-cyanophenyl)-3-aminobenzenesulfonamide (265.6mg, 86.0%). N-(2-chloro-5-cyanophenyl)-3-aminobenzenesulfonamide (265.6 mg), benzaldehyde (456.3 mg), and sodium methoxide (232.3 mg) were dissolved in methanol (20 mL) and protected with nitrogen. The reaction solution was stirred at 65°C for 3 h. After the reaction was completed, it was cooled to room temperature and a methanol solution of sodium borohydride (162.7 mg) was added at 0°C. The reaction solution was stirred at 65°C for 9 h, and ice water was added to the reaction solution. The solvent was dried and extracted with ethyl acetate. The organic phase was washed with saturated NaCl, dried over anhydrous Na2SO4, and purified by silica gel column chromatography with petroleum ether / ethyl acetate (4:1) to obtain compound I-16 (41.1 mg, 12.1%). 1H NMR (300MHz, Chloroform-d) δ7.92 (d, J=1.8Hz, 1H), 7.42 (s, 1H), 7.39 (s, 1H), 7.37-7.32 (m, 5H), 7.30 (d , J=1.6Hz, 1H), 7.26 (d, J=8.0Hz, 1H), 7.21-7.16 (m, 2H), 7.13 (s, 1H), 6.88 (d, J=8.2Hz, 1H), 4.35 (s, 2H).

[0074] Example 17: Preparation of Compound I-17

[0075] N-(2-chloro-5-cyanophenyl)-3-bromobenzenesulfonamide was synthesized in the same manner as compound I-1. N-(2-chloro-5-cyanophenyl)-3-bromobenzenesulfonamide (400.5 mg), aniline (129.6 mg), palladium acetate (26.1 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (443 mg), and cesium carbonate (1.13 g) were dissolved in 1,4-dioxane (20 mL). The reaction solution was stirred at 100°C for 10 h. After the reaction was completed, it was cooled to room temperature, the solvent was dried and extracted with ethyl acetate, the organic phase was washed with saturated NaCl, dried over anhydrous Na2SO4, and purified by silica gel column chromatography with petroleum ether / ethyl acetate (6:1) to obtain compound I-17 (102.4 mg, 23%). 1 H NMR (300MHz, Chloroform-d) δ7.96 (d, J=1.8Hz, 1H), 7.45 (d, J=8.3Hz, 1H), 7.41 (t, J=2.1Hz, 1H), 7.38-7.34 (m, 2H), 7.34-7.3 0 (m, 2H), 7.18 (ddd, J=7.8, 2.4, 1.4Hz, 1H), 7.14 (s, 1H), 7.12-7.08 (m, 1H), 7.06 (q, J=1.2, 0.7Hz, 1H), 7.03 (t, J=1.1Hz, 1H).

[0076] Example 18: Preparation of Compound I-18

[0077] N-(2-chloro-5-cyanophenyl)-3-aminobenzenesulfonamide was synthesized by the same method as compound I-16. N-(2-chloro-5-cyanophenyl)-3-aminobenzenesulfonamide (265.6 mg), benzoyl chloride (120.9 mg), and TEA (261 mg) were dissolved in dichloromethane (20 mL), and the reaction solution was stirred at room temperature for 6 h. After the reaction, the solvent was dried and extracted with ethyl acetate. The organic phase was washed with saturated NaCl, dried over anhydrous Na2SO4, and purified by silica gel column chromatography with petroleum ether / ethyl acetate (4:1) to obtain compound I-18 (159.4 mg, 45%). 1 H NMR (300MHz, Chloroform-d) δ9.27 (s, 1H), 8.03 (t, J=2.3Hz, 1H), 7.95 (dt, J=8.0, 1.2Hz, 2H), 7.86 (ddd, J=7.7, 2.2, 1. 2Hz, 1H), 7.65 (ddd, J=8.4, 2.2, 1.2Hz, 1H), 7.59-7.51 (m, 2H), 7.54-7.49 (m, 1H), 7.51-7.43 (m, 3H), 7.46-7.40 (m, 1H).

[0078] Example 19: Preparation of Compound I-19

[0079] Compound I-19 (84.9 mg, 17.9%) was synthesized in the same manner as compound I-1, using 2-chloro-3-fluoro-5-cyanoaniline (0.33 g) instead of 2-chloroaniline. 1 H NMR (300MHz, Chloroform-d) δ7.73 (ddd, J=8.8, 2.2, 1.2Hz, 1H), 7.65 (d, J=2.2Hz, 1H), 7.43 (dd, J=8.0, 2.1Hz, 1H), 7.40 (dd, J=8.8, 7.2Hz, 1H), 7.12 (t , J=2.2Hz, 1H), 6.80 (ddd, J=7.3, 2.2, 1.2Hz, 1H), 3.23 (ddd, J=5.3, 4.0, 1.0 Hz, 4H), 2.84-2.76 (m, 2H), 2.57 (ddd, J=12.5, 5.2, 4.2Hz, 2H), 2.29 (s, 3H).

[0080] Example 20: Preparation of Compound I-20

[0081] Compound I-20 (92.8 mg, 18.8%) was synthesized in the same manner as compound I-1, using 2,4-dichloro-5-cyanoaniline (0.37 g) instead of 2-chloroaniline. 1H NMR (300MHz, Chloroform-d) δ7.73 (ddd, J=8.8, 2.2, 1.2Hz, 1H), 7.60 (s, 1H), 7.50 (s, 1H), 7.40 (dd, J =8.8, 7.2Hz, 1H), 7.12 (t, J = 2.3Hz, 1H), 6.80 (ddd, J = 7.3, 2.2, 1.2Hz, 1H), 3.23 (ddd, J = 5.3, 4.0, 1.0 Hz, 4H), 2.84-2.76 (m, 2H), 2.57 (ddd, J=12.4, 5.2, 4.2Hz, 2H), 2.29 (s, 3H).

[0082] Example 21: Preparation of Compound I-21

[0083] Using 2-chloro-5-cyanoaniline (0.30 g) instead of 2-chloroaniline and 3-methoxy-5-bromobenzenesulfonyl chloride (0.56 g) instead of 3-bromobenzenesulfonyl chloride, the same method as compound I-1 was used to synthesize compound I-21 (88.4 mg, 18.1%). 1 HNMR (300MHz, Chloroform-d) δ7.57 (d, J=1.9Hz, 1H), 7.49 (d, J=8.4Hz, 1H), 7.43 (dd, J=8.5, 1.9Hz, 1H), 7.23 (d, J=4.4Hz, 1H), 7.06 ( t, J=2.2Hz, 1H), 6.55 (t, J=2.2Hz, 1H), 3.82 (s, 3H), 3.33-3.22 (m, 4H), 2.87 (ddd, J=12.3, 5.5, 3.9Hz, 2H), 2.58 (ddd, J=12.3, 5.4, 4.0 Hz, 2H), 2.29 (s, 3H).

[0084] Example 22: Preparation of Compound I-22

[0085] Using 2-chloro-5-cyanoaniline (0.30 g) instead of 2-chloroaniline and 3-bromo-4-cyanobenzenesulfonyl chloride (0.55 g) instead of 3-bromobenzenesulfonyl chloride, compound I-22 (86.8 mg, 18.0%) was synthesized in the same manner as compound I-1. 1H NMR (300MHz, Chloroform-d) δ7.78 (dd, J=9.2, 2.2Hz, 1H), 7.73 (d, J=9.2Hz, 1H), 7.57 (d, J=1.9Hz, 1H), 7.49 (d, J=8.4Hz, 1H ), 7.46-7.39 (m, 2H), 3.22-3.10 (m, 4H), 2.87 (ddd, J=12.4, 5.9, 3.6Hz, 2H), 2.58 (ddd, J=12.4, 5.9, 3.5Hz, 2H), 2.29 (s, 3H).

[0086] Example 23: Preparation of Compound I-23

[0087] Using cyclopropylboronic acid (0.49 g) instead of methylpiperazine, the compound I-23 (86.0 mg, 24.1%) was synthesized in the same manner as compound I-1. 1 H NMR (300MHz, Chloroform-d) 7.81-7.77 (m, 1H), 7.76 (ddd, J=8.4, 2.2, 1.3Hz, 1H), 7.47-7.40 (m, 1H), 7.39-7.33 (m, 2H), 7.30 (dd, J=7. 8, 1.5Hz, 1H), 7.22 (td, J=7.5, 1.3Hz, 1H), 6.92 (ddd, J=8.6, 7.2, 1.6Hz, 1H), 2.55-2.47 (m, 1H), 1.04-0.92 (m, 2H), 0.85-0.73 (m, 2H).

[0088] Example 24: Preparation of Compound I-24

[0089] Using cyclobutylboronic acid (0.58 g) instead of methylpiperazine, compound I-24 (94.4 mg, 25.3%) was synthesized in the same manner as compound I-1. 1 H NMR (300MHz, Chloroform-d) 7.76 (ddd, J=8.4, 2.2, 1.3Hz, 1H), 7.73 (td, J=2.2, 0.7Hz, 1H), 7.43 (dd, J=8.4, 7.7Hz, 1 H), 7.36 (dd, J=8.0, 1.3Hz, 1H), 7.34-7.27 (m, 2H), 7.22 (td, J=7.5, 1.3Hz, 1 H), 6.92 (ddd, J=8.6, 7.2, 1.6Hz, 1H), 2.90-2.82 (m, 1H), 1.96-1.65 (m, 6H).

[0090] Example 25: Preparation of Compound I-25

[0091] Using cyclopentylboronic acid (0.66 g) instead of methylpiperazine, the compound I-25 (88.0 mg, 22.6%) was synthesized in the same manner as compound I-1. 1 H NMR (300MHz, Chloroform-d) δ7.79-7.71 (m, 2H), 7.43 (d, J=16.1Hz, 1H), 7.36 (dd, J=8.1, 1.3Hz, 1H), 7.30 (dt, J=7.7, 1.4Hz, 2H), 7.22 (td, J=7.5, 1.3Hz, 1H), 6.92 (ddd, J=8.6, 7.2, 1.6Hz, 1H), 2.97-2.90 (m, 1H), 1.85-1.73 (m, 2H) , 1.71 (ddd, J=8.0, 5.5, 3.9Hz, 2H), 1.68 (d, J=1.6Hz, 2H), 1.69-1.61 (m, 2H).

[0092] Example 26: Preparation of Compound I-26

[0093] Using cyclohexylboronic acid (0.74 g) instead of methylpiperazine, the compound I-26 (84.8 mg, 20.9%) was synthesized in the same manner as compound I-1. 1 H NMR (300MHz, Chloroform-d) δ7.79-7.71 (m, 2H), 7.43 (d, J=16.1Hz, 1H), 7.36 (dd, J=8.0, 1.3Hz, 1H), 7.30 (dt, J=8.1, 1.5Hz, 2H), 7.22 (td, J=7.5, 1.3Hz, 1H), 6.92 (ddd, J=8.6, 7.2, 1.6Hz, 1H), 2.72-2.64 (m, 1H), 1.86-1.76 (m, 2H), 1.73-1.36 (m, 8H).

[0094] Pharmacological experiments and results of some compounds of the present invention

[0095] 1. The inhibitory effect of the compounds of the present invention on LDHA protein was tested by spectrophotometry.

[0096] In order to evaluate the inhibitory effect of the series of inventive compounds on LDHA protein, the present invention uses spectrophotometry to test the inhibitory effect of the series of compounds on LDHA protein. The test method is as follows: the test equipment is a 96-well plate. After the mother solution of the tested series of compounds is diluted with buffer to obtain 6 different concentrations, 2μL of compound DMSO solution, 194μL LDHA protein and sodium pyruvate protein buffer are added to each well. After incubation at 37°C for 5 minutes, 4μL NADH solution is added to each well, and then the absorbance at 340nm is immediately detected using a Spectra Max i3x multifunctional microplate reader. The test results are analyzed using Graphpad Prism. 5μL LDHA is added to 95μL protein buffer (200mM Tris HCl, pH=7.4, 100μM EDTA and 0.01% Tween-20) to prepare a protein buffer solution.

[0097] Table 1. NADH: Lactate dehydrogenase 1 inhibitory activity of the example compounds

[0098]

[0099]

[0100] Where a represents IC 50 Less than 300nM; b indicates IC 50 :300~400nM; c represents IC 50 :400~600nM; d represents IC 50 Greater than 600nM.

[0101] As shown in Table 1, the N-phenylbenzenesulfonamide compounds of the present invention have strong NADH: lactate dehydrogenase 1 inhibitory activity.

[0102] 2. MTT colorimetric method (MTT assay) to test the cytotoxicity of some compounds of the present invention on human pancreatic cancer cell line MiaPaCa-2.

[0103] The experimental method is as follows: tumor cells are inoculated in a 96-well plate and then placed in an incubator to wait for the cells to adhere. All compounds are prepared into 10mM stock solutions, and the compounds are diluted to 100μM with cell culture medium, and then three-fold dilutions are performed to obtain six concentrations in succession. Take 100μL of compound solution and add it to each well of the 96-well plate. Incubate the 96-well plate at 37°C for 72 hours, then add 20μL of 5mg / ml MTT solution to each well and incubate in the incubator for 4 hours. After the incubation is completed, the solution in each well is aspirated and removed, 150μL of PBS buffer is added to each well to wash twice, 150μL of DMSO solution is added to each well, shake on a shaker for 10 minutes, and the absorbance at 570nm is detected. The test results are analyzed using GraphpadPrism.

[0104] Table 2. Cytotoxicity of Example Compounds to Mia PaCa-2 Cells

[0105]

[0106]

[0107] Where a represents IC 50 :0~500nM;b represents IC 50 :500~900nM; c represents IC 50 Greater than 900nM.

[0108] 3. Lactic acid level test Testing the inhibitory activity of some compounds of the present invention on the production of lactic acid

[0109] The ability of compounds to inhibit lactate production in tumor cells was evaluated to confirm whether these compounds exert their anti-tumor effects by inhibiting LDHA activity. The experimental method was as follows: MiaPaCa-2 tumor cells (2x10 5 / well) in a six-well plate and incubated overnight. The compounds were then added to the corresponding wells and incubated at 37°C for 4 hours. Finally, the cells in the culture medium were collected from each well, broken by ultrasonic treatment, and centrifuged for 10 minutes (10000rpm) to remove cell debris. The lactic acid content in the culture medium was analyzed using a lactate assay kit (Solarbio).

[0110] Table 3. Lactate inhibition levels of the example compounds

[0111]

[0112] Where a represents IC 50 :0~5μM; b represents IC 50 :5~10μM; c represents IC 50 :10~15μM; d represents IC50 Greater than 20μM.

[0113] It can be seen that the N-phenylbenzenesulfonamide compounds of the present invention have anti-tumor activity, cytotoxicity and lactic acid inhibition activity comparable to those of the positive control GNE-140, and can be used to prepare anti-tumor drugs.

Claims

1. An N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the compound is shown in general formula (I): Wherein P1 represents phenyl, 3-7 membered cycloalkyl, 3-7 membered heterocycloalkyl; the 3-7 membered heterocycloalkyl is a 3-7 membered ring containing two or three elements of C, N, and O; X represents -(CH2) n -NH-, amide or absent, n is selected from 0, 1, 2, 3, 4; R1 represents hydrogen or C1-C6 alkyl; R2 represents one or more of hydrogen, monosubstituted or polysubstituted halogen, cyano, C1-4 alkoxy, trifluoromethyl; R3 represents hydrogen, C1-C6 alkyl, acetyl, trifluoroacetyl, oxalyl, tert-butyloxycarbonyl, or C1-C4 chain-linked carboxyl; R4 represents hydrogen, monosubstituted or polysubstituted halogen, cyano, C 1～4 Alkoxy.

2. The N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: P1 represents phenyl, 3- to 6-membered cycloalkyl, or 3- to 6-membered heterocycloalkyl; the 3- to 6-membered heterocycloalkyl is a 3- to 6-membered ring containing two or three of C, N, and O.

3. The N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: P1 represents a phenyl group, a 3- to 6-membered cycloalkyl group, a piperazinyl group, or a morpholinyl group.

4. The N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: R2 represents hydrogen, monosubstituted or polysubstituted halogen, cyano, C 1～4 One, two or three of alkoxy and trifluoromethyl.

5. The N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound is selected from the following compounds:

6. A method for preparing the N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof as claimed in claim 1, characterized in that: When X is absent and P1 is a nitrogen-containing aliphatic ring, compound IV and The target compound I is obtained by reaction at a reaction temperature of 80 to 130° C., wherein R1, R2, R3, R4, and P1 are defined as in claim 1. R3 is substituted piperazinyl or morpholinyl: When X is -(CH2) n -NH-, n is selected from 0, 1, 2, 3, 4, compound IV and The target product II is obtained by reaction at a reaction temperature of 80 to 130° C., wherein R1, R2, R3, R4, and P1 are defined as in claim 1. -(CH2) substituted by R3 n -phenyl, 3-7 membered cycloalkyl-(CH2) n -; n is selected from 0, 1, 2, 3, 4: When X is an amide group, compound V and The target compound III is obtained by reaction at room temperature, wherein R1, R2, R3, R4, and P1 are defined as in claim 1, P1 substituted with R3 and COCl:

7. A pharmaceutical composition, characterized in that Contains the N-phenylbenzenesulfonamide compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 and a pharmaceutically acceptable carrier.

8. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7 in the preparation of an LDH enzyme inhibitor.

9. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating malignant tumors.

10. The use according to claim 9, characterized in that: The malignant tumor is pancreatic cancer.

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