Stilbenes and-1 protein degraders, methods of making, and uses thereof

Abstract

The application discloses a new compound with an antitumor effect shown in formula I, a preparation method and application of the compound. Experiments show that the compound can inhibit proliferation of various tumor cells, mechanism research shows that the compound induces degradation of acidic nuclear matrix DNA binding protein (And-1), inhibits DNA double-strand damage repair of tumor cells, and is expected to become a new type of antitumor and tumor radiotherapy and chemotherapy sensitizing drug.

Description

Technical Field

[0001] This invention relates to a novel small molecule compound with antitumor activity, a method for preparing the compound, and its application in the preparation of antitumor or tumor radiotherapy and chemotherapy sensitizers. Background Technology

[0002] The following background information related to this invention is provided to aid in understanding the invention, but should not be considered as prior art. All cited publications are referenced in their entirety.

[0003] Compared with normal cells, tumor cells exhibit a variety of abnormal biological characteristics, including persistent proliferation signaling, induction of angiogenesis, metabolic reprogramming, immune escape, inflammation, and genomic instability. Studies have shown that an overactive DNA damage repair system promotes tumor cell invasion and metastasis (Cancer Discov. 2017; 7(7): 675-693). Therefore, inhibiting the DNA repair system has always been a hot topic in anti-tumor drug research. With the clinical potential of PARP inhibitors being validated, the development of new drugs targeting the tumor DNA damage repair system is expected to achieve breakthroughs in expanding anti-tumor indications, overcoming radiotherapy resistance, and cisplatin resistance (Nat Rev Genet. 2017; 18(10): 613-623).

[0004] Acidic nucleoplasmic DNA-binding protein 1 (And-1) contains a WD40 repeat sequence (WD-repeat) and a high-mobility histone box domain (HMG-box), possessing characteristics of both the WD40 and HMG protein families. Studies have shown that the WD40 repeat domain of And-1 is essential for preventing the accumulation of DNA single-strand and double-strand damage and cell cycle arrest (Nat Commnn. 2018; 9(1): 3091). In addition, Chen Y et al. found that when DNA is damaged, And-1 interacts with CtIP, recruiting CtIP to the DNA damage site, promoting DNA end excision and DNA damage repair (Nucleic Acids Res. 2017; 45(5): 2516-2530). Clinical studies have shown that And-1 immunostaining positivity is associated with poor prognosis in patients with non-small cell lung cancer and esophageal cancer (Clin Cancer Res. 2010; 16: 226-39.). Multivariate analysis showed that And-1 is an independent prognostic factor for esophageal cancer. Inhibition of And-1 expression with small interfering RNA effectively suppressed the growth of lung and esophageal cancer cells (Clin Cancer Res. 2010; 16: 226-39.). MicroRNA-494-dependent And-1 inhibition reduced epithelial-mesenchymal transition, tumor growth, and metastasis in cholangiocarcinoma cells (Digestive and LiverDisease. 2019; 51: 397-411.). Therefore, And-1 can serve as a potential target for anti-tumor drugs. Developing small-molecule And-1 protein degraders is of great significance.

[0005] In our previous study, we screened two highly efficient and specific And-1 inhibitors, bardoxifene acetate and CH3, using an And-1 luciferase reporter gene high-throughput screening platform. Specifically, bardoxifene and CH3 block And-1 polymerization by interacting with the And-1 WD40 domain, promoting its interaction with the E3 ligase Cullin4B (CUL4B), ultimately leading to its ubiquitination and degradation. Therefore, a key characteristic of this class of inhibitors is that they are also degradative agents of the And-1 protein. Summary of the Invention

[0006] The present invention aims to address the deficiencies of the prior art by providing a new method for preparing and applying a stilbene compound with antitumor activity, thereby solving the technical problem of the lack of a similar compound in the prior art.

[0007] Another technical problem that this invention aims to solve is that there are currently no clinical candidate drugs that exert anti-tumor effects by degrading the And-1 protein;

[0008] Another technical problem to be solved by the present invention is to provide the application of the said compound in antitumor drugs and tumor radiotherapy and chemotherapy sensitizers. To achieve the above technical objectives, the present invention adopts the following technical solution:

[0009] A compound represented by Formula I:

[0010]

[0011] in:

[0012] n = 0 or 1

[0013] R is selected from piperazine, phenyl, furanyl, thiophene, pyridinyl, pyrazolyl, imidazolyl, triazolyl, pyrimidinyl, indolyl, pyridazine, benzopyrazinyl, benzimidazolyl, benzimidazolone, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, inzolyl, indololinone, naphridinyl, quinolinyl, quinolinone, dihydroquinolinone, oxo-dihydroquinolinone, isoquinolinyl, isoquinolinone, wherein each R is optionally substituted with 1 to 3 substituents, which are independently selected from hydrogen, trifluoromethyl, halogen, C1-C3 alkyl, C1-C3 alkoxy, 4-7 membered heterocyclic, hydroxyl, amino, acetamido, carboxyl, heteroaryl, ester.

[0014] Preferably, the compound or a pharmaceutically usable salt thereof is selected from:

[0015]

[0016]

[0017] The present invention provides a pharmaceutical composition comprising the above-described compound or a pharmaceutically usable salt thereof, and a pharmaceutically acceptable carrier, including a diluent.

[0018] Meanwhile, the present invention provides the application of the above-mentioned compounds or their pharmaceutically usable salts, and the above-mentioned pharmaceutical compositions in the preparation of And-1 degrading agent-related drugs.

[0019] Preferably, the above-mentioned compound or its pharmaceutically usable salt, or the above-mentioned pharmaceutical composition, is a drug for anti-tumor or tumor radiotherapy and chemotherapy sensitization.

[0020] Meanwhile, the present invention provides a method for preparing the above-mentioned compound, comprising the following steps:

[0021] 1) Compound A reacts with concentrated hydrochloric acid and paraformaldehyde to give compound B;

[0022] 2) Compound C is obtained by reacting compound B with triphenylphosphine in an aprotic solvent;

[0023] 3) Compound D is obtained by reacting compound C with 3,4-dichlorobenzaldehyde under strongly alkaline conditions;

[0024] 4) An imine intermediate was prepared by reacting compound D with RNH2, and then reduced to obtain compound I-1-1-27;

[0025] The structure of the compound is as follows:

[0026]

[0027] The R in RNH2 is defined as described above.

[0028] Preferably, the aprotic solvent for the reaction in step 2) includes any one or a combination of several of the following: acetone, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, ethyl acetate, methyl tert-butyl ether, and dimethyl sulfoxide.

[0029] Preferably, the reaction in step 3) is carried out under strongly alkaline conditions, including any one or a combination of several of sodium methoxide, sodium ethoxide, sodium tert-butoxide, sodium isopropoxide, butyllithium, sodium hydroxide, lithium hydroxide, and potassium hydroxide.

[0030] Preferably, the reducing agent in step 4) includes any one or a combination of several of sodium borohydride, sodium cyanoborohydride, sodium triacetoxyborooxide, and lithium aluminum hydride.

[0031] Unless otherwise specified, the technical terms related to the above technical solutions shall follow the definitions below.

[0032] The term "alkyl" refers to a straight-chain or branched hydrocarbon group having a specified number of carbon atoms; therefore, for example, the terms "C1-C4 alkyl" and "C1-C4 alkyl" are used herein. 10 "Alkyl" refers to an alkyl group having at least one and at most four or ten carbon atoms. Examples of such branched or straight-chain alkyl groups used in this invention include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl, n-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, and n-decyl, and branched analogs of the following five n-alkanes.

[0033] When the term "heterocyclic alkyl" is used, it refers to a non-aromatic heterocycle containing a specified number of ring atoms, which is saturated or has one or more degrees of unsaturation, and contains one or more heteroatoms selected from O, S, or N. Such a ring may optionally be fused to one or more other "heterocyclic" or cycloalkyl groups. Examples of "heterocyclic" groups include monocyclic non-aromatic heterocyclic groups such as azirrobutylyl (e.g., 1-azirrobutylyl, 2-azirrobutylyl, 3-azirrobutylyl), pyrroliyl (e.g., 1-pyrroliyl, 2-pyrroliyl), piperidinyl (e.g., piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl), morpholinyl (e.g., morpholino), thiomorpholinyl (e.g., thiomorpholino), piperazinyl (e.g., 1-piperazinyl, 2-piperazinyl, 3-piperazinyl), hexazinyl, etc. Hexamethyleniminyl (e.g., hexamethyleneimin-1-yl), oxazolidinyl (e.g., oxazolidin-2-yl), thiazolyl (e.g., thiazolyl-2-yl), imidazolyl (e.g., imidazolyl-2-yl, imidazolyl-3-yl), oxazolinyl (e.g., oxazolinyl-2-yl), thiazolyl (e.g., thiazolyl-2-yl), imidazolyl (e.g., imidazolyl-2-yl, imidazolyl-3-yl), dioxacyclopentenyl (e.g., ... 1,3-dihydrohexacyclopenten-4-yl), dioxacyclopentyl (e.g., 1,3-dioxacyclopentan-4-yl), dihydrooxadiazolyl (e.g., 4,5-dihydro-1,2,4-oxadiazol-3-yl), pyranyl (e.g., 4-pyranyl), tetrahydropyranyl (e.g., 2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl), thiopyranyl (e.g., 4-thiopyranyl), tetrahydrothiopyranyl (e.g., 2-tetrahydrothiopyranyl, 3-tetrahydrothiopyranyl, 4-tetrahydropyranyl), thiopyranyl (e.g., 4-thiopyranyl), tetrahydrothiopyranyl (e.g., 2-tetrahydrothiopyranyl, 3-tetrahydrothiopyranyl, 4-tetrahydrothiopyranyl), 4-tetrahydrothiopyranyl -Tetrahydrothiopyranyl), tetrahydrofuranyl (e.g., tetrahydrofuran-3-yl, tetrahydrofuran-2-yl), pyrazolyl (e.g., pyrazolidine-1-yl, pyrazolidine-3-yl), pyrazolinyl (e.g., pyrazolidine-1-yl), tetrahydropyrimidinyl (e.g., tetrahydropyrimidin-1-yl), dihydrotriazolyl (e.g., 2,3-dihydro-1H-1,2,3-triazol-1-yl), tetrahydrotriazolyl (e.g., 2,3,4,5-tetrahydro-1H-1,2,3-triazol-1-yl), etc.;Fused non-aromatic heterocyclic groups, such as dihydroindolyl (e.g., 2,3-dihydro-1H-indol-1-yl), dihydroisoindolyl (e.g., 1,3-dihydro-2H-isoindol-2-yl), dihydrobenzofuranyl (e.g., 2,3-dihydro-1-benzofuran-5-yl), dihydrobenzodioxanedienyl (e.g., 2,3-dihydro-1,4-benzodioxanedienyl), dihydrobenzodioxaneheptatrienyl (e.g., 3,4-dihydro-2H-1,5-dioxaneheptatrienyl), tetrahydrobenzofuranyl (e.g., 4,5,6,7-tetrahydrobenzofuranyl). Hydrogen-1-benzofuran-3-yl), chromenyl (e.g., 4H-chromen-2-yl, 2H-chromen-3-yl), dihydrochromenyl (e.g., 3,4-dihydro-2H-chromen-2-yl), dihydroquinolinyl (e.g., 1,2-dihydroquinolin-4-yl), tetrahydroquinolinyl (e.g., 1,2,3,4-tetrahydroquinolin-4-yl), dihydroisoquinolinyl (e.g., 1,2-dihydroisoquinolin-4-yl), tetrahydroisoquinolinyl (e.g., 1,2,3,4-tetrahydroisoquinolin-4-yl), dihydrophthalazinyl (e.g., 1,4-dihydrophthalazin-4-yl), etc.

[0034] The term "aryl" refers to an aromatic group containing 5-14 ring atoms, with at least one ring possessing a conjugated π-electron system. This includes aromatic rings with all carbon atoms, aromatic heterocycles, and fused or biaromatic rings, and may contain substituents. Aryl groups can contain 1-6 substituents. A heteroaromatic ring or aromatic heterocycle refers to a group containing 5-14 ring atoms, of which 1-4 heteroatoms are aromatic ring atoms, and the remaining ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, nitrogen, and selenium atoms. Suitable aromatic heterocycles include furan, thiophene, pyridine, pyrrolidine, pyrrolidines with low-carbon alkyl substituents on the nitrogen atom, pyridine nitrides, pyrimidines, pyrazines, imidazoles, and other similar heterocycles, all of which may contain substituents.

[0035] The terms "arbitrary substitution" or "substitution" refer to groups having 1-4 different substituents, which can be: low-carbon alkyl, low-carbon aryl, low-carbon aralkyl, low-carbon cyclic alkyl, low-carbon heterocyclic alkyl, hydroxyl, low-carbon alkoxy, low-carbon aryloxy, polyhaloalkoxy, arylalkoxy, low-carbon heteroaryl, low-carbon heteroarylepoxy, low-carbon heteroarylalkyl, low-carbon heteroarylalkoxy, azide, amino, halogen, low-carbon alkylthiol, oxy, low-carbon acylalkyl. Low carbon number carboxylic acid ester group, carboxylic acid, amide group, nitro group, low carbon number acyloxy group, low carbon number amine alkyl group, low carbon number alkylamine aryl group, low carbon number alkylaryl group, low carbon number alkylamine alkyl group, low carbon number alkoxyaryl group, low carbon number arylamine group, low carbon number arylalkylamine group, sulfonyl group, low carbon number amide alkylaryl group, low carbon number amide aryl group, low carbon number hydroxyalkyl group, low carbon number haloalkyl group, low carbon number alkylamine alkyl acid group, low carbon number urea alkyl group, cyano group, low carbon number alkoxyalkyl group, low carbon number polyhaloalkyl group, low carbon number arylalkoxyalkyl group.

[0036] "Substituted aryl" and "substituted heteroaryl" refer to aromatic rings or heteroaryl groups with 1 to 6 substituents. These substituents can be low-carbon alkyl groups, low-carbon alkoxy groups, low-carbon polyhaloalkyl groups, halogens, hydroxyl groups, and amino groups. Attached Figure Description

[0037] Figure 1 Effects of the compound on And-1 protein expression (A549 cells);

[0038] Figure 2 Effects of compound I-15 on clone formation in A549 and H460 cells. Detailed Implementation

[0039] The compounds and their preparation in this invention can be better illustrated by the following examples. These examples should not be construed as limiting the invention, and variations of these compounds, whether now known or developed in the future, should also be considered within the scope of this invention and protected.

[0040] The specific embodiments of the present invention will be described in detail below. To avoid excessive and unnecessary details, well-known structures or functions will not be described in detail in the following embodiments.

[0041] The approximate language used in the following embodiments is for quantitative expression, indicating that a certain degree of variation in quantity is permissible without changing the basic function. Therefore, the values ​​corrected using terms such as "approximately," "around," etc., are not limited to the exact value itself. In some embodiments, "approximately" indicates that the value being corrected is allowed to vary within a range of plus or minus ten percent (±10%); for example, "approximately 100" represents any value between 90 and 110. Furthermore, in the expression "approximately from the first value to the second value," both the first and second values ​​are corrected simultaneously. In some cases, the approximate language may be related to the accuracy of the measuring instrument.

[0042] Unless otherwise defined, the technical and scientific terms used in the following embodiments have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0043] Example 1: 5-(chloromethyl)-2-hydroxybenzaldehyde (B)

[0044]

[0045] A solution of paraformaldehyde (24.40 g, 0.20 mol) in concentrated hydrochloric acid (80 mL) was stirred at room temperature for 10 minutes. Salicylaldehyde (17.95 g, 0.60 mol) was added dropwise over 30 minutes. The reaction mixture was stirred at room temperature for 24 hours, yielding a white precipitate. The precipitate was filtered, washed with cold water, and dried to obtain the crude product. The crude product was recrystallized from n-hexane (310 mL) to give white needle-like crystals (16.80 g, 49.4%). 1 H NMR (400MHz, DMSO-d6) δ10.91 (s, 1H), 10.27 (s, 1H), 7.72 (d, J=2.4Hz, 1H), 7.59 (dd, J=8.5, 2.4Hz, 1H), 7.03 (d, J=8.5Hz, 1H), 4.76 (s, 2H).

[0046] Example 2 (3-formyl-4-hydroxybenzyl) triphenylphosphonium chloride (C)

[0047]

[0048] 5-(chloromethyl)-2-hydroxybenzaldehyde (15.50 g, 91.2 mmol) and triphenylphosphine (30.98 g, 118.1 mmol) were dissolved in 220 mL of dry acetonitrile. The reaction mixture was stirred at 80 °C for 3 hours under nitrogen protection. The acetonitrile was evaporated to dryness under reduced pressure, and the remaining triphenylphosphine was washed away with petroleum ether (100 mL × 3). The mixture was filtered to obtain a white powder (38.52 g, 97.8%).

[0049] Example 3(E)-5-(3,5-dichlorostyryl)-2-hydroxybenzaldehyde(D)

[0050]

[0051] Under nitrogen protection, sodium (0.19 g, 8.26 mmol) was dissolved in anhydrous ethanol (30 mL). The phosphine salt from the previous step (1.20 g, 2.78 mmol) was added to this sodium ethanol solution until the reaction solution turned deep yellow. Then, 3,4-dichlorobenzaldehyde (0.57 g, 3.26 mmol) was added. The reaction solution was stirred at 75 °C for 3 hours. The reaction was stopped, cooled to room temperature, and clarified first with water, then became turbid. The pH was adjusted to acidic with dilute hydrochloric acid, precipitating a yellow solid. The solid was filtered, dried, and recrystallized twice from tetrahydrofuran to obtain yellow crystals (0.387 g, 44.4%).

[0052] Example 4 Synthesis of Compound I-1

[0053] In a 25 mL single-necked flask, compound D (0.334 mmol, 1.0 eq) and ethanolamine (0.344 mmol, 1.03 eq) were dissolved in 10 mL of anhydrous ethanol. The mixture was stirred at 78 °C for 3 hours to obtain a brownish-red reaction solution. The solution was cooled to room temperature, and TLC was used to monitor the complete consumption of the reactants. 3 mL of anhydrous tetrahydrofuran and sodium borohydride (0.676 mmol, 2.0 eq) were added directly to the system, and the mixture was stirred at room temperature for 1 hour until the intermediate was completely converted. The reaction was quenched with water, extracted with ethyl acetate, and the organic phase was washed three times with saturated brine. Column chromatography yielded a yellow solid, 80% yield. HRMS(ESI)[MH] - =336.0567; 1 H NMR (300MHz, DMSO-d6) δ7.82 (d, J=1.5Hz, 1H), 7.55 (dt, J=8.4, 5.1Hz, 2H), 7.42-7.31 (m, 2H), 7.26 (d, J=16.4H z, 1H), 7.00 (d, J=16.4Hz, 1H), 6.73 (d, J=8.2Hz, 1H), 3.87 (s, 2H), 3.51 (t, J=5.6Hz, 2H), 2.62 (t, J=5.6Hz, 2H). 13 C-NMR (101MHz, DMSO-d6): δ158.54, 139.16, 131.89, 131.48, 131.16, 129.15, 127. 93, 127.67, 127.57, 127.40, 126.50, 124.78, 122.78, 116.23, 60.20, 50.86, 50.72.

[0054] Following the method described above, the compounds of Examples 5 to 30 were synthesized as follows:

[0055] Sodium 5(E)-2-((cyclopentylamino)methyl)-4-(3,4-dichlorostyryl)phenol(I-2)

[0056] Yellow solid, 75% yield; HRMS(ESI)[MH] - =360.0930; 1H NMR (300MHz, DMSO-d6): δ7.81 (d, J=1.7Hz, 1H), 7.55 (dt, J=8.5, 5.1Hz, 2H), 7.33 (d, J=7.7Hz, 2H), 7.26 (d, J= 16.5Hz, 1H), 7.00 (d, J=16.4Hz, 1H), 6.70 (d, J=8.3Hz, 1H), 3.85 (s.2H), 3.08-2.97 (m, 1H), 1.86-1.35 (m, 8H). 13 C NMR (101MHz, DMSO-d6) δ158.99, 139.17, 131.89, 131.50, 131.15, 129.13, 127.92, 12 7.48, 127.37, 127.33, 126.49, 124.85, 122.72, 116.32, 58.79, 49.89, 32.28, 23.85.

[0057] Example 6 (E)-4-(3,4-dichlorostyryl)-2-((isopropylamino)methyl)phenol(I-3)

[0058] Yellow solid, 77% yield; HRMS(ESI)[MH] - =334.0775; 1 H NMR (300MHz, DMSO-d6) δ7.81 (d, J=1.7Hz, 1H), 7.54 (dt, J=8.5, 5.1Hz, 2H), 7.33 (d, J=8.4Hz, 2H), 7.26 (d, J=16.5Hz, 1H), 7.00 (d, J=16.4Hz, 2H), 6.70 (d, J=8.0Hz, 1H), 3.88 (s, 2H), 2.75 (dt, J=12.6, 6.3Hz, 1H), 1.06 (d, J=6.3Hz, 6H). 13 C NMR (101MHz, DMSO-d6) δ159.13, 139.18, 131.89, 131.51, 131.15, 129.12, 12 7.91, 127.42, 127.27, 126.48, 124.91, 122.68, 116.35, 48.78, 47.87, 22.50.

[0059] Example 7

[0060] (E)-4-(3,4-dichlorostyryl)-2-(((furan-2-ylmethyl)amino)methyl)phenol(I-4):

[0061] Yellow solid, yield 66%; HRMS(ESI)[MH] - =372.0566; 1 H NMR (300MHz, DMSO-d6) δ7.83 (d, J=1.5Hz, 1H), 7.64-7.51 (m, 3H), 7.44-7.31 (m, 2H), 7.27 (d, J=16.4Hz, 1H), 7. 01 (d, J=16.4Hz, 1H), 6.76 (d, J=8.3Hz, 1H), 6.46-6.40 (m, 1H), 6.31 (d, J=3.0Hz, 1H), 3.80 (s, 2H), 3.73 (s, 2H). 13 C NMR (101MHz, DMSO-d6) δ157.93, 153.71, 142.63, 139.16, 131.89, 131.49, 131.16, 129.16, 127 .95, 127.78, 127.76, 127.31, 126.52, 125.13, 122.88, 116.13, 110.84, 107.77, 49.51, 44.81.

[0062] Example 8

[0063] Methyl(E)-3-((5-(3,4-dichlorostyryl)-2-hydroxybenzyl)amino)propanoate(I-5):

[0064] White solid, 73% yield; HRMS(ESI)[MH] - =378.0674; 1 H NMR (300MHz, DMSO-d6) δ7.82 (d, J=1.6Hz, 1H), 7.55 (dt, J=8.4, 5.0Hz, 2H), 7.34 (dd, J=13.6, 5.3Hz, 2H), 7.26 (d, J=16 .4Hz, 1H), 7.01 (d, J=16.4Hz, 1H), 6.74 (d, J=8.3Hz, 1H), 3.81 (s, 2H), 3.60 (s, 3H), 2.78 (t, J=6.7Hz, 2H), 2.54 (s, 1H). 13C NMR (101MHz, DMSO-d6) δ172.78, 158.07, 139.16, 131.89, 131.48, 131.16, 129.16, 127.9 3, 127.70, 127.64, 127.35, 126.51, 125.20, 122.86, 116.15, 51.77, 50.38, 44.38, 34.31.

[0065] Example 9

[0066] (E)-4-(3,4-dichlorostyryl)-2-(((4-methylpiperazin-1-yl)amino)methyl)phenol(I-6):

[0067] White solid, yield 72%; HRMS(ESI)[MH] - =390.0975; 1 H NMR (300MHz, DMSO) δ11.47 (s, 1H), 7.90 (s, 1H), 7.84 (d, J=1.5Hz, 1H), 7.65-7.52 (m, 3H), 7.44 (dd, J=8.5, 1.8Hz, 1H), 7.31 (d, J=16.4Hz, 1H), 7.04 (d, J=16.4Hz, 1H), 6.88 (d, J=8.4Hz, 1H), 3.14 (s, 4H), 2.53 (s, 4H), 2.24 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ157.24, 138.91, 138.76, 131.93, 131.19, 130.95, 129.45, 12 8.30, 128.14, 128.08, 127.81, 126.69, 123.77, 120.47, 116.94, 54.11, 51.04, 45.92.

[0068] Example 10

[0069] (E)-2-(((4-(1H-imidazol-1-yl)phenyl)amino)methyl)-4-(3,4-dichlorostyryl)phenol(I-7)

[0070] Yellow solid, 70% yield; HRMS(ESI)[MH] - =434.0840; 1H NMR (400MHz, DMSO) δ9.94 (s, 1H)-7.97 (s-1H), 7.82 (d, J=1.8Hz, 1H)-7.56 (d, J=8.4Hz, 1H), 7.54-7.46 (m, 3H)-7.36 (dd, J=8.4, 2.0Hz, 1H), 7.27 ( t, J=11.9Hz, 3H), 7.02 (s, 1H), 6.94 (d, J=16.4Hz, 1H), 6.88 (d, J=8.3Hz, 1H), 6.69 (d, J=8.9Hz, 2H), 6.33 (t, J=5.8Hz, 1H), 4.24 (d, J=5.7Hz, 2H). 13 C NMR (101MHz, DMSO-d6) δ156.07, 148.48, 139.03, 135.82, 131.89, 131.61, 131.12, 129.54, 129.21, 128.00, 127.98, 127.76, 126.78, 126.60, 126.29, 122.98, 122.49, 118.87, 115.84-112.99, 41.95.

[0071] Example 11

[0072] (E)-4-(3,4-dichlorostyryl)-2-(((4-morpholinophenyl)amino)methyl)phenol(I-8):

[0073] Yellow solid, yield 76%; HRMS(ESI)[MH] - =454.1208; 1 H NMR (300MHz, DMSO-d6) δ9.86 (s, 1H), 7.81 (d, J = 1.7Hz, 1H), 7.52 (dt, J = 13.1, 5 .1Hz, 3H), 7.33 (dd, J=8.3, 2.0Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 6.93 (d, J=16.4H z, 1H), 6.85 (d, J=8.3Hz, 1H), 6.74 (d, J=8.9Hz, 2H), 6.55 (d, J=8.9Hz, 2H), 5.5 9 (t, J=5.4Hz, 1H), 4.15 (d, J=4.9Hz, 2H), 3.76-3.62 (m, 4H), 2.96-2.81 (m, 4H). 13C NMR (101MHz, DMSO-d6) δ156.06, 143.48, 142.95, 139.06, 131.89, 131.68, 131.11, 129.17, 127.97, 127.91, 127.69, 126.96, 126.63, 126.54, 122.86, 118.01, 115.73, 113.66, 66.78, 50.97, 42.80.

[0074] Example 12 (E)-4-(3,4-dichlorostyryl)-2-((phenylamino)methyl)phenol(I-9):

[0075] White solid, 75% yield; HRMS(EsI)[M] - =369.0568; 1 H NMR (400MHz, DMSO-d6) δ9.89 (s, 1H), 7.82 (s, 1H), 7.60-7.45 (m, 3H), 7.35 (dd, J=8.3, 1.6Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 7.06 (t, J=7.8Hz, 2 H), 6.93 (d, J=16.4Hz, 1H), 6.86 (d, J=8.3Hz, 1H), 6.60 (d, J=7.9Hz, 2H), 6.52 (t, J=7.2Hz, 1H), 6.02 (t, J=5.7Hz, 1H), 4.19 (d, J=5.6Hz, 2H). 13 C NMR (101MHz, DMSO-d6) δ156.04, 149.35, 139.04, 131.89, 131.64, 131.11, 129.31, 129.19, 127.97, 127.93, 127.76, 126.68, 126.64, 126.57, 122.90, 116.18, 115.77, 112.68, 41.96.

[0076] Example 13(E)-4-(3,4-dichlorostyryl)-2-((pyridin-3-ylamino)methyl)phenol(I-10):

[0077] White solid, yield 72%; HRMS(ESI)[M+H] + =371.0725; 1H NMR (400MHz, DMSO-d6) δ9.94 (s, 1H), 8.02 (s, 1H), 7.81 (d, J=1.9Hz, 1H), 7.75 ( d, J=3.6Hz, 1H), 7.56 (d, J=8.4Hz, 1H), 7.51 (dd, J=8.5, 1.9Hz, 1H), 7.47 (d, J= 1.9Hz, 1H), 7.36 (dd, J=8.4, 2.1Hz, 1H), 7.25 (d, J=16.4Hz, 1H), 7.06 (dd, J=8. 2, 4.6Hz, 1H), 6.98-6.85 (m, 3H), 6.30 (t, J=5.7Hz, 1H), 4.22 (d, J=5.8Hz, 2H). 13 C NMR (101MHz, DMSO-d6) δ156.10, 145.32, 139.02, 137.28, 135.90, 131.89, 131.53, 131.11, 129 .22, 128.01, 127.98, 127.85, 126.93, 126.59, 125.95, 124.08, 123.00, 117.99, 115.88, 41.50.

[0078] Example 14 (E)-4-((5-(3,4-dichlorostyryl)-2-hydroxybenzyl)amino)benzoic acid (I-11):

[0079] White solid, yield 74%; HRMS(ESI)[MH] - =412.0512; 1 H NMR (400MHz, DMSO-d6) δ12.01 (s, 1H), 9.95 (s, 1H), 7.82 (d, J=1.8Hz, 1H), 7.6 6 (d, J=8.8Hz, 2H), 7.56 (d, J=8.4Hz, 1H), 7.52 (dd, J=8.5, 1.9Hz, 1H), 7.42 (d, J=1.7Hz, 1H), 7.38 (dd, J=8.4, 2.0Hz, 1H), 7.25 (d, J=16.4Hz, 1H), 6.94 (d, J=1 6.4Hz, 1H), 6.87 (t, J=6.8Hz, 2H), 6.62 (dJ=8.8Hz, 2H), 4.26 (d, J=5.6Hz, 2H). 13C NMR (101MHz, DMSO-d6) δ167.96, 156.06, 153.06, 139.01, 131.88, 131.58, 131.51, 131.11, 129 .22, 128.01, 127.99, 127.86, 126.91, 126.62, 125.78, 123.03, 117.47, 115.88, 111.48, 41.48.

[0080] Example 15

[0081] (E)-4-(3,4-dichlorostyryl)-2-(((3-(trifluoromethyl)benzyl)amino)methyl)phenol(I-12):

[0082] Yellow solid, yield 67%; HRMS(ESI)[MH] - =450.0652; 1 H NMR (300MHz, DMSO-d6) δ7.82 (d, J=1.7Hz, 1H), 7.74 (s, 1H), 7.71-7.49 (m, 5H), 7.41 (s, 1H), 7.35 (d, J=8. 3Hz, 1H), 7.27 (d, J=16.4Hz, 1H), 7.00 (d, J=16.4Hz, 1H), 6.78 (d, J=8.3Hz, 1H), 3.84 (s, 2H), 3.80 (s, 2H). 13 C NMR (101MHz, DMSO-d6) δ157.77, 141.87, 139.14, 132.70, 131.90, 131.50, 131.16, 129.97, 129.73, 129.65, 129.34, 129.18 , 129.03, 127.94, 127.82, 127.76, 127.33, 126.50, 126.18, 125.36, 125.00, 124.01, 123.47, 122.88, 116.10, 51.72, 49.47.

[0083] Example 16

[0084] (E)-4-(3,4-dichlorostyryl)-2-(((pyridin-4-ylmethyl)amino)methyl)phenol(I-13)

[0085] Yellow solid, yield 68%; HRMS(ESI)[MH] - =383.0726;1 1H NMR (400 MHz, DMSO-d6) δ 8.53 (d, J = 3.3 Hz, 2H), 7.83 (d, J = 1.9 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.53 (dd, J = 8.5, 1.9 Hz, 1H), 7.43 (d, J = 1.9 Hz, 1H), 7.38 (d, J = 5.6 Hz, 2H), 7.34 (dd, J = 8.4, 2.1 Hz, 1H), 7.28 (d, J = 16.4 Hz, 1H), 7.01 (d, J = 16.4 Hz, 1H), 6.78 (d, J = 8.3 Hz, 1H), 3.78 (d, J = 2.1 Hz, 4H). 13 13C NMR (101 MHz, DMSO-d6) δ 157.59, 149.98, 149.39, 139.14, 131.90, 131.50, 131.16, 129.18, 127.95, 127.84, 127.30, 126.53, 125.47, 123.54, 122.90, 116.06, 51.15, 49.31.

[0086] Example 17

[0087] (E)-4-(3,4-dichlorostyryl)-2-(((4-(dimethylamino)phenyl)amino)methyl)phenol (I-14)

[0088] Yellow solid, yield 62%; HRMS (ESI) [M-H] - = 411.1043; 1 1H NMR (300 MHz, DMSO-d6) δ 9.86 (s, 1H), 7.81 (d, J = 1.6 Hz, 1H), 7.53 (dt, J = 12.1, 5.1 Hz, 3H), 7.33 (dd, J = 8.3, 1.9 Hz, 1H), 7.24 (d, J = 16.4 Hz, 1H), 6.93 (d, J = 16.4 Hz, 1H), 6.84 (d, J = 8.3 Hz, 1H), 6.62 (d, J = 8.9 Hz, 2H), 6.55 (d, J = 8.9 Hz, 2H), 5.39 (s, 1H), 4.14 (s, 2H), 2.70 (s, 6H). 13C NMR (101MHz, DMSO-d6) δ156.10, 143.47, 141.57, 139.07, 131.88, 131.69, 131.12, 129.15 , 127.96, 127.88, 127.68, 127.08, 126.61, 126.55, 122.83, 115.71, 114.09, 43.15, 42.21.

[0089] Example 18

[0090] (E)-4-(3,4-dichlorostyryl)-2-(((4-(4-methylpiperazin-1-yl)phenyl)amino)methyl)phenol(I-15)

[0091] Yellow solid, yield 69%; HRMS(ESI)[MH] - =466.1460; 1 H NMR (300MHz, DMSO-d6) δ9.92 (s, 1H), 7.81 (s, 1H), 7.53 (dd, J=18.0, 11.7Hz, 3H), 7.33 (d, J=8.3Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 6.89 (dd, J=19.2, 12.3Hz, 2H), 6.73 (d, J=8.8Hz, 2H), 6.54 (d, J=8.8Hz, 2H), 5.56 (s, 1H), 4.15 (s, 2H), 2.98-2.84 (m, 4H), 2.42 (m, 4H), 2.19 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ156.12, 143.25, 142.99, 139.08, 131.88, 131.70, 131.11, 129.15, 127.97, 12 7.85, 127.66, 126.98, 126.60, 126.55, 122.82, 118.25, 115.74, 113.65, 55.37, 50.52, 46.25, 42.83.

[0092] Example 19

[0093] (E)-2-(((1H-indol-5-yl)amino)methyl)-4-(3,4-dichlorostyryl)phenol(I-16)

[0094] Brownish-red solid, yield 65%; HRMS(ESI)[MH] -=407.0729; 1 H NMR (400MHz, DMSO-d6) δ10.63 (s, 1H), 9.91 (s, 1H), 7.80 (d, J=1.9Hz, 1H), 7.52 (dt, J=8.5, 5.2Hz, 3H), 7.33 (dd, J=8.3, 2.1Hz, 1H), 7.23 (d, J=16.4Hz , 1H), 7.13 (t, J=6.4Hz, 2H), 6.93 (d, J=16.3Hz, 1H), 6.85 (d, J=8.3Hz, 1H) , 6.63 (dd, J=4.5, 2.2Hz, 2H), 6.18-6.13 (m, 1H), 5.43 (s, 1H), 4.22 (s, 2H). 13 C NMR (101MHz, DMSo-d6) δ156.20, 142.72, 139.07, 131.87, 131.72, 131.10, 130.18, 129.14, 128.84, 127.97, 127.89, 127.67, 127.20, 126.59, 126.54, 125.12, 122.82, 115.73, 112.08, 111.96, 101.22, 100.49, 43.74.

[0095] Example 20

[0096] (E)-4-(3,4-dichlorostyryl)-2-(((pyridin-3-ylmethyl)amino)methyl)phenol(I-17)

[0097] White solid, yield 72%; HRMS(ESI)[MH] - =383.0727; 1 H NMR (400MHz, DMSO-d6) δ8.55 (d, J=1.1Hz, 1H), 8.52-8.44 (m, 1H), 7.82 (d, J=1.8Hz, 1H), 7.77 (d, J=7.8Hz, 1H), 7.59 (d, J=8.4Hz, 1H), 7.53 (dd, J=8.4, 1.8Hz, 1H), 7.45-7.32 (m, 3H), 7.28 (d, J=16.4Hz, 1H), 7.01 (d, J=16.4Hz, 1H), 6.77 (d, J=8.3Hz, 1H), 3.80 (s, 2H), 3.77 (s, 2H). 13CNMR (101MHz, DMSO-d6) δ157.80, 149.99, 148.64, 139.15, 136.29, 135.62, 131.90, 131.51, 131.16 , 129.18, 127.95, 127.84, 127.79, 127.30, 126.52, 125.29, 123.91, 122.90, 116.12, 49.75, 49.52.

[0098] Example 21

[0099] (E)-N-(4-((5-(3,4-dichlorostyryl)-2-hydroxybenzyl)amino)phenyl)acetamide(I-18)

[0100] Yellow solid, 70% yield; HRMS(ESI)[MH] - =425.0830; 1 H NMR (400MHz, DMSO-d6) δ9.87 (s, 1H), 9.52 (s, 1H), 7.82 (d, J=1.8Hz, 1H), 7.53 (dt, J=8.5, 5.1Hz, 2H), 7.47 (d, J=1.9Hz, 1H), 7.34 (dd, J=8.4, 2.1Hz, 1 H), 7.29-7.21 (m, 3H), 6.93 (d, J = 16.4Hz, 1H), 6.85 (d, J = 8.3Hz, 1H), 6.54 ( d, J=8.9Hz, 2H), 5.82 (t, J=5.9Hz, 1H), 4.17 (d, J=5.8Hz, 2H), 1.95 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ167.66, 156.05, 145.54, 139.06, 131.88, 131.64, 131.11, 129.21, 129.18 , 127.99, 127.93, 127.77, 126.72, 126.70, 126.57, 122.90, 121.30, 115.75, 112.61, 42.39, 24.15.

[0101] Example 22

[0102] (E)-4-(3,4-dichlorostyryl)-2-(((3-(4-methylpiperazin-1-yl)phenyl)amino)methyl)phenol(I-19)

[0103] Yellow solid, yield 66%; HRMS(ESI)[MH] - =466.1464; 1 H NMR (300MHz, DMSO-d6) δ9.97 (s, 1H), 7.78 (s, 1H), 7.50 (q, J=8.2Hz, 3H), 7.34 (d, J=8.1Hz, 1H), 7.22 (d, J=16.3 Hz, 1H), 6.91 (t, J=12.5Hz, 3H), 6.15 (m, 3H), 5.80 (s, 1H), 4.22 (s, 2H), 3.03 (s, 4H), 2.41 (s, 4H), 2.19 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ156.04, 152.40, 150.06, 139.06, 131.89, 131.64, 131.13, 129.63, 129.18, 127.97, 12 7.93, 127.87, 126.91, 126.80, 126.55, 122.86, 115.72, 104.60, 104.39, 100.40, 55.17, 48.72, 46.19, 42.03.

[0104] Example 23

[0105] (E)-4-(3,4-dichlorostyryl)-2-(((2-methoxy-4-(4-methylpiperazin-1-yl)phenyl)amino)methyl)phenol(I-20)

[0106] Yellow solid, yield 56%; HRMS(ESI)[MH] - =496.1572; 1 H NMR (300MHz, CDCl3) δ7.55 (d, J=1.7Hz, 1H), 7.32 (dt, J=8.1, 5.1Hz, 4H), 7.02 (d, J=16.3Hz, 1H), 6.82 (m, 3H), 6.55 ( d, J=2.2Hz, 1H), 6.45 (dd, J=8.5, 2.1Hz, 1H), 4.39 (s, 2H), 3.85 (s, 3H), 3.21-3.06 (m, 4H), 2.63 (m, 4H), 2.38 (s, 3H). 13C NMR (101MHz, DMSO-d6) δ156.17, 147.76, 143.40, 139.07, 132.45, 131.88, 131.64, 131.10, 129.17, 127.98, 1 27.90, 126.94, 126.69, 126.58, 122.90, 115.81, 110.65, 108.52, 102.03, 55.82, 55.37, 50.51, 46.23, 43.26.

[0107] Example 24

[0108] (E)-4-(3,4-dichlorostyryl)-2-(((4-((4-methylpiperazin-1-yl)methyl)phenyl)amino)methyl)phenol(I-21)

[0109] Yellow solid, yield 67%; HRMS(ESI)[MH] - =480.1622; 1 H NMR (400MHz, DMSO-d6) δ10.04 (s, 1H), 7.80 (d, J = 1.8Hz, 1H), 7.56 (d, J = 8.4Hz, 1H), 7.50 (dd, J = 8.5, 1.8Hz, 1H), 7.46 (d, J = 1.7Hz, 1H), 7.33 (dd, J = 8.3, 1.9Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 7.00-6.84 (m, 4H), 6.54 (d, J=8.4 Hz, 2H), 5.95 (s, 1H), 4.18 (s, 2H), 3.24 (s, 2H), 2.27 (s, 8H), 2.11 (s, 3H). 13 C NMR (101MHz, DMSO-d6) δ156.19, 148.37, 139.06, 131.88, 131.69, 131.11, 130.20, 129.15, 127.95, 1 27.79, 127.69, 126.71, 126.55, 125.56, 122.79, 115.78, 112.38, 62.45, 55.23, 52.90, 46.23, 42.14.

[0110] Example 25

[0111] (E)-4-(3,4-dichlorostyryl)-2-(((3-fluoro-4-(4-methylpiperazin-1-yl)phenyl)amino)methyl)phenol(I-22)

[0112] Yellow solid, yield 63%; HRMS(ESI)[MH] - =484.1371; 1 H NMR (400MHz, DMSO-d6) δ10.02 (s, 1H), 7.82 (d, J = 1.9Hz, 1H), 7.57 (d, J = 8.4Hz, 1H), 7.52 (dd, J=8.5, 1.9Hz, 1H), 7.44 (dd, J=10.9, 2.0Hz, 1H), 7.35 (dd, J=8.4, 2.1Hz, 1H), 7.25 (d, J=16.4 Hz, 1H), 6.93 (d, J=16.4Hz, 1H), 6.86 (d, J=8.3Hz, 1H), 6.84-6.77 (m, 1H), 6.36 (ddd, J=11.8, 11.1, 2.5Hz, 2H), 6.00 (s, 1H), 4.14 (d, J=3.1Hz, 2H), 2.81 (s, 4H), 2.41 (s, 4H), 2.19 (s, 3H). 13 CNMR (101MHz, DMSO-d6) δ158.01, 156.11, 155.61, 146.13, 146.03, 139.05, 131.89, 131.62, 131.12, 129.78, 129.68, 129.19, 127.98, 1 27.91, 127.80, 126.76, 126.58, 126.45, 122.91, 121.02, 120.98, 115.82, 108.21, 108.19, 100.97, 100.74, 55.42, 51.46, 46.30, 42.21.

[0113] Example 26

[0114] (E)-4-(3,4-dichlorostyryl)-2-(((4-(4-isopropylpiperazin-1-yl)phenyl)amino)methyl)phenol(I-23)

[0115] Yellow solid, 71% yield; HRMS(ESI)[MH] - =494.1782; 1H NMR (400MHz, DMSO-d6) δ9.87 (s, 1H), 7.81 (d, J = 1.8Hz, 1H), 7.56 (d, J = 8.4Hz, 1H), 7.51 (dd, J = 8.5, 1 .9Hz, 1H), 7.48 (d, J=1.9Hz, 1H), 7.33 (dd, J=8.4, 2.1Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 6.93 (d, J=16.4 Hz, 1H), 6.84 (d, J=8.3Hz, 1H), 6.72 (d, J=8.9Hz, 2H), 6.53 (d, J=8.9Hz, 2H), 5.55 (s, 1H), 4.15 (d, J=1 .7Hz, 2H), 2.95-2.80 (m, 4H), 2.65 (dt, J=12.5, 6.2Hz, 1H), 2.54-2.50 (m, 4H), 0.99 (d, J=6.5Hz, 6H). 13 C NMR (101MHz, DMSO-d6) δ156.07, 143.23, 143.12, 139.07, 131.89, 131.69, 131.11, 129.17, 127.98, 127.9 0, 127.68, 127.00, 126.61, 126.55, 122.85, 118.24, 115.72, 113.65, 54.18, 51.01, 48.72, 42.85, 18.70.

[0116] Example 27

[0117] (E)-4-(3,4-dichlorostyryl)-2-(((4-(4-(methylsulfonyl)piperazin-1-yl)phenyl)amino)methyl)phenol(I-24)

[0118] Yellow solid, yield 64%; HRMS(ESI)[MH] - =530.1074; 1H NMR (300MHz, DMSO-d6) δ9.87 (s, 1H), 7.82 (s, 1H), 7.53 (dd, J=19.2, 12.0Hz, 3H), 7.34 (d, J=8.2Hz, 1H), 7.25 (d, J=16.4Hz, 1H), 6.93 (d, J=16. 4Hz, 1H), 6.85 (d, J=8.3Hz, 1H), 6.78 (d, J=8.5Hz, 2H), 6.56 (d, J=8.5Hz , 2H), 5.66(s, 1H), 4.16(s, 2H), 3.21(m, 4H), 2.99(m, 4H), 2.91(s, 3H). 13 CNMR (101MHz, DMSO-d6) δ156.05, 144.00, 142.27, 139.05, 131.89, 131.68, 131.12, 129.18, 127.97, 1 27.92, 127.70, 126.91, 126.63, 126.56, 122.88, 119.21, 115.74, 113.54, 50.65, 46.07, 42.67, 34.17.

[0119] Example 28

[0120] (E)-4-(3,4-dichlorostyryl)-2-(((4-(4,4-difluoropiperidin-1-yl)phenyl)amino)methyl)phenol(I-25)

[0121] Yellow solid, 67% yield: HRMS(ESI)[MH] - =487.1170; 1 H NMR (300MHz, DMSO-d6) δ9.86 (s, 1H), 7.81 (d, J = 1.7Hz, 1H), 7.56 (d, J = 8.4Hz, 1H), 7.50 (dd, J=13.9, 1.8Hz, 2H), 7.34 (dd, J=8.4, 1.9Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 6 .93 (d, J=16.4Hz, 1H), 6.85-6.75 (m, 3H), 6.55 (d, J=8.8Hz, 2H), 5.64 (t, J=5.2Hz, 1H), 4.15 (d, J=4.8Hz, 2H), 3.12-2.99 (m, 4H), 2.03 (ddd, J=19.9, 14.0, 5.7Hz, 4H). 13C NMR (101MHz, DMSO-d6) δ156.05, 143.86, 141.81, 139.05, 131.89, 131.67, 131.11, 129.18, 127.97, 127.91, 127.70, 126.92, 126.65, 126.55, 123.25, 122.87, 119.50, 115.73, 113.56, 48.49, 48.44, 48.39, 42.69, 34.16, 33.94, 33.72.

[0122] Example 29

[0123] (E)-4-(3,4-dichlorostyryl)-2-(((4-(4-methyl-1,4-diazepan-1-yl)phenyl)amino)methyl)phenol(I-26)

[0124] Yellow solid, yield 59%; HRMS(ESI)[MH] - =480.1607; 1 H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 7.81 (d, J=1.9Hz, 1H), 7.53 (dt, J=11.4, 5.4Hz, 3H ), 7.32 (dd, J=8.4, 2.1Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 6.93 (d, J=16.4Hz, 1H), 6.84 (d, J=8.3Hz, 1H), 6.52 (d, J=3.5Hz, 4H), 5.29 (s, 1H), 4.13 (s, 2H), 3.39-3.34 (m, 2H), 3.29 (t, J=6.2Hz, 2H), 2.59-2.53(m, 2H), 2.47-2.38(m, 2H), 2.23(s, 3H), 1.88-1.78(m, 2H). 13 C NMR (101MHz, DMSO-d6) δ156.19, 141.95, 140.12, 139.09, 131.88, 131.72, 131.12, 129.15.127.97, 127.86, 127. 67, 127.17, 126.64, 126.53, 122.81, 115.72, 114.72, 113.50, 58.03, 56.92, 49.42, 48.84, 46.61, 43.56, 27.70.

[0125] Example 30

[0126] (E)-4-(3,4-dichlorostyryl)-2-(((4-(3,5-dimethylpiperazin-1-yl)phenyl)amino)methyl)phenol(I-27)

[0127] Yellow solid, yield 57%; HRMS(ESI)[MH] - =480.1598; 1 H NMR (400MHz, DMSO-d6) δ9.87 (s, 1H), 7.81 (d, J = 1.9Hz, 1H), 7.56 (d, J = 8.4Hz, 1H), 7.51 (dd, J = 8.5, 1.9Hz, 1H), 7.48 (d, J=2.0Hz, 1H), 7.33 (dd, J=8.4, 2.1Hz, 1H), 7.24 (d, J=16.4Hz, 1H), 6.93 (d, J=16.4Hz, 1H), 6.84 (d, J=8 .3Hz, 1H), 6.71 (d, J=8.9Hz, 2H), 6.54 (s, 2H), 5.53 (s, 1H), 4.14 (s, 2H), 3.65-3.56 (m, 1H), 3.20 (dd, J=11.0, 1 .9Hz, 2H), 2.82 (ddd, J=9.4, 6.3, 2.7Hz, 2H), 1.96 (t, J=10.7Hz, 2H), 1.80-1.73 (m, 1H), 0.97 (d, J=6.3Hz, 6H). 13 C NMR (101MHz, DMSO-d6) δ156.08, 143.27, 143.07, 139.06, 131.88, 131.68, 131.12, 129.15, 127.97, 127.87, 1 27.68, 127.00, 126.63, 126.55, 122.83, 118.34, 115.70, 113.68, 67.49, 57.98, 50.75, 42.85, 25.60, 19.97.

[0128] Example 31 Immunoblot Detection of And-1 Expression

[0129] Experimental Procedure: A549 cells in logarithmic growth phase were digested, centrifuged, resuspended, and seeded at approximately 200,000 cells per well in 6-well plates. Cells were lysed with an appropriate volume of RIPA protease lysis agent and a protease inhibitor (PMSF), followed by sonication to ensure complete lysis. Protein concentration was determined and quantified using the BCA method. After adding SDS loading buffer, the cells were boiled for denaturation. The samples were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the proteins were transferred to a PVDF membrane. The membrane was blocked for 60 min at room temperature in PBST solution containing 5% skim milk, and then incubated with primary antibody at 4°C for 12–18 hours. After washing three times with PBST for 5 min each time, the membrane was incubated with secondary antibody at room temperature for 90 min, followed by three times with PBST for 5 min each time. The membrane was then placed in a pre-chilled gel imaging system, ECL chromogenic solution was added, and images were formed. The final images were analyzed using ImageJ software. All samples were processed in triplicate.

[0130] Experimental results are as follows Figure 1 As shown, the results indicate that, compared with the control group, the compounds of this invention can significantly induce the expression of And1 protein in A549 cells.

[0131] Example 32: Compound Inhibition of Cell Clonal Formation Experiment

[0132] Experimental Procedure: A549 cells in logarithmic growth phase were harvested, digested, centrifuged, resuspended, and seeded at approximately 400 cells per well in 12-well plates. Cells were then treated with the PARP-1 inhibitor olaparib and compound I-15, with the medium containing DMSO or the compound changed every 3 days. After 2 weeks of incubation, cells were washed with PBS, fixed with 4% paraformaldehyde for 20 min, stained with crystal violet for 20 min, and finally photographed. Colonies with ≥50 cells were counted. All samples were obtained in triplicate.

[0133] Experimental results are as follows Figure 2 As shown, the results indicate that in A549 and H460 cells, the combination of the compound of the present invention with Olaparib can significantly inhibit cell colony formation and has a synthetic lethal effect.

Claims

1. The following compounds and their pharmaceutically usable salts: 。 2. A method for preparing the compound of claim 1 and its pharmaceutically usable salt, comprising the following steps: 1) Compound A reacts with concentrated hydrochloric acid and paraformaldehyde to give compound B; 2) Compound C is obtained by reacting compound B with triphenylphosphine in an aprotic solvent; 3) Compound D is obtained by reacting compound C with 3,4-dichlorobenzaldehyde under strong alkaline conditions; wherein the strong alkaline is selected from any one or a combination of several of sodium methoxide, sodium ethoxide, sodium tert-butoxide, sodium isopropoxide, butyllithium, sodium hydroxide, lithium hydroxide, and potassium hydroxide. 4) An imine intermediate is prepared by reacting compound D with RNH2, and then reduced to obtain the compound of claim 1; wherein, .

3. The preparation method according to claim 2, wherein: Step 2) The aprotic solvent for the reaction is selected from any one or a combination of several of the following: acetone, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane, ethyl acetate, methyl tert-butyl ether, and dimethyl sulfoxide.

4. The preparation method according to claim 2, wherein: Step 4) The reduction is performed using a reducing agent, wherein the reducing agent is selected from any one or a combination of several of sodium borohydride, sodium cyanoborohydride, and sodium triacetoxyborohydride.

5. A composition, characterized in that, It comprises the compound of claim 1 and its pharmaceutically usable salt, as well as a pharmaceutically acceptable carrier.

6. The use of the compound of claim 1 and its pharmaceutically usable salt in the preparation of an antitumor drug.

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