Application of a bromodomain protein 4 active compound based on a water network strategy

Abstract

The present invention discloses an application of a bromodomain protein 4 active compound based on a water network strategy. The bromodomain protein 4 active compound can be used to prepare a drug that inhibits the activity of bromodomain protein 4, and can be used to prepare a pharmaceutically acceptable salt. In addition, the bromodomain protein 4 active compound and the pharmaceutically acceptable salt prepared therefrom can also be used to prepare a bromodomain protein 4 inhibitor and the pharmaceutically acceptable salt prepared therefrom, as well as a pharmaceutical composition for preparing a prostate cancer cell proliferation inhibitor, an anti-prostate tumor drug, and a pharmaceutical composition for treating bromodomain protein 4-related diseases. The bromodomain protein 4 inhibitor prepared based on the bromodomain protein 4 active compound having obvious inhibitory activity of the present invention is applied to the treatment of diseases related to bromodomain protein 4, has high reliability, is easy to promote and popularize, and utilizes the potential advantages of the bromodomain protein 4 active compound as a drug or pharmaceutical composition, opening up a new approach for the treatment of related diseases.

Description

Technical Field

[0001] The present invention relates to an application of an active compound, relates to the technical field of biochemistry, and particularly relates to an application of a bromodomain protein 4 active compound based on a water network strategy. Background Art

[0002] Prostate cancer is a common and fatal cancer in men, and its incidence is currently showing a significant upward trend. For patients with localized (early-stage) prostate cancer, radical prostatectomy and radiotherapy can achieve good therapeutic effects. However, due to the lack of obvious clinical symptoms and hidden disease progression in the early stages of prostate cancer, coupled with the lack of widespread screening, many patients present with advanced metastatic disease by the time they seek medical attention. Radical prostatectomy and chemotherapy are unlikely to achieve optimal results at this stage, necessitating the use of endocrine therapy. For patients with hormone-sensitive advanced prostate cancer, endocrine therapy is the mainstay of treatment, including surgical castration, medical castration, and the use of anti-androgen drugs, primarily androgen receptor (AR) antagonists. For patients with castration-resistant prostate cancer (CRPC), AR antagonists are an important treatment option.

[0003] Although AR antagonists (antiandrogens) have a long history of use in the treatment of prostate cancer, and were initially used in combination with chemical castration drugs, gonadotropin-releasing hormone (LHRH) analogs, as a supplement to androgen deprivation therapy (ADT), to block the aggravation of disease symptoms caused by a short-term increase in testosterone levels in patients in the early stages of medical castration, various drug-resistant mutations of AR itself arise as the disease progresses. These mutations significantly reduce the efficacy of existing drugs, leading to the disease becoming incurable in the later stages.

[0004] To this end, new drug targets for prostate cancer have gradually attracted the attention of scientists, and bromodomain-containing protein 4 (BRD4) is one of them. As an epigenetic protein, BRD4 not only participates in gene recruitment and transcription, but also has the ability to bind to multiple proteins such as MYC. In CRPC cells, it acts as a coactivator, binding to the dimeric AR, initiating transcription and completing the expression of cancer genes. It also participates in mediating the process of enhanced chromatin accessibility driven by dysregulated AR signaling, forming a positive feedback loop.

[0005] As the relationship between targeting BRD4 and treating CRPC becomes clear, more and more BRD4 small molecule inhibitors are being used to treat CRPC. Although the clinical trial results of ZEN-3694 showed that it has a relatively good efficacy in patients with CRPC who are resistant to enzalutamide, as many as 18.7% of the patients tested experienced grade 3 or higher toxic reactions, which shows that it is limited by drug safety in the treatment of CRPC. In addition, some BD1 / BD2 selective small molecule inhibitors have also been found to have certain effects on the treatment of CRPC, such as ABBV-744, which has less platelet and intestinal toxicity than its non-selective homologs. These new findings indicate that BRD4 inhibitors can effectively solve the drug resistance problem of existing AR-targeted antagonists for the treatment of CRPC.

[0006] A review of the research status of BRD4 inhibitors for CRPC both domestically and internationally reveals that the development of a new generation of inhibitors targeting the BRD4 protein with novel backbone structures, high affinity, and high selectivity remains a research priority. With the increasing problem of an aging population, there is a huge clinical need for this. Anti-CRPC inhibitors targeting the BRD4 protein could overcome the resistance limitations of traditional AR antagonists, a research area that still presents a significant clinical gap; the development of novel BRD4 inhibitors for CRPC is urgently needed. Summary of the Invention

[0007] To address the problems presented in the prior art, the present invention provides an application of a bromodomain protein 4 active compound based on a water network strategy. The present invention aims to provide a compound with bromodomain protein 4 inhibitory activity for application in the preparation of bromodomain protein 4 inhibitors and anti-prostate tumor drugs.

[0008] The technical solution adopted in the present invention is:

[0009] The applications of the bromodomain protein 4 active compound of the present invention are as follows:

[0010] Computer-aided drug design was used to discover lead compounds targeting bromodomain protein 4, and then virtual screening based on molecular docking was performed on multiple small molecule compound three-dimensional structure databases. In this process, the receptor protein water network strategy was applied emphatically to obtain the top 1000 compounds (the lower the energy, the higher the score). Then, cell proliferation experiments using the classic prostate cancer cell line LNCaP with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric method (MTT method) and the kit EPIgeneous TMBinding Domain Kit A (Cisbio brand) was used to investigate the binding of compounds to the bromodomain protein 4BRD4, and ultimately 21 representative active compounds were screened.

[0011] The invention relates to an application of the bromodomain protein 4 active compound in the preparation of a drug for inhibiting the activity of bromodomain protein 4.

[0012] The structural formula of the bromodomain protein 4 active compound is specifically one of the following structural formulas:

[0013]

[0014] Structural formula (1) is N-(3-chlorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0015] Structural formula (2) is N-(4-fluorophenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide.

[0016] Structural formula (3) is N-(2-bromo-4-methylphenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide.

[0017] Structural formula (4) is N-(3-fluoro-4-methylphenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide.

[0018] Structural formula (5) is N-(2,4-difluorophenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide.

[0019] Structural formula (6) is N-(4-bromo-2-methylphenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide.

[0020] Structural formula (7) is N-(4-fluorobenzyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0021] Structural formula (8) is N-(2-chloro-4-fluorobenzyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0022] Structural formula (9) is N-(4-fluorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0023] Structural formula (10) is 4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)-N-(3-(trifluoromethyl)phenyl)butanamide.

[0024] Structural formula (11) is N-(2-bromo-4-methylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0025] Structural formula (12) is N-(2,4-difluorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0026] Structural formula (13) is N-(5-chloro-2-methoxyphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0027] Structural formula (14) is N-(2-fluorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0028] Structural formula (15) is N-(3-methoxyphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0029] Structural formula (16) is N-(5-chloro-2,4-dimethoxyphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0030] Structural formula (17) is N-cycloheptyl-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0031] Structural formula (18) is N-(4-ethylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0032] Structural formula (19) is N-(4-isopropylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0033] Structural formula (20) is N-(4-chloro-2-methoxy-5-methylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0034] Structural formula (21) is N-(3-cyanophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

[0035] The 21 active compounds showed good effects in anti-prostate tumor experiments at the protein and cell levels.

[0036] Application of the bromodomain protein 4 active compound in the preparation of pharmaceutically acceptable salts.

[0037] The pharmaceutically acceptable salt is hydrochloride, phosphate, sulfate, acetate, maleate, citrate, benzenesulfonate, toluenesulfonate, fumarate or tartrate.

[0038] Application of the bromodomain protein 4 active compound and the pharmaceutically acceptable salt prepared therefrom in the preparation of bromodomain protein 4 inhibitors.

[0039] The invention relates to an application of the bromodomain protein 4 active compound and the pharmaceutically acceptable salt prepared therefrom in the preparation of a prostate cancer cell proliferation inhibitor.

[0040] The invention relates to the use of the bromodomain protein 4 active compound and the pharmaceutically acceptable salt prepared therefrom in the preparation of anti-prostate tumor drugs.

[0041] The invention relates to an application of the bromodomain protein 4 active compound and the pharmaceutically acceptable salt prepared therefrom in the preparation of a pharmaceutical composition for treating bromodomain protein 4 diseases.

[0042] The therapeutic pharmaceutical composition specifically includes a pharmaceutically acceptable excipient, diluent or carrier, such as syrup, gum arabic and starch.

[0043] The therapeutic pharmaceutical composition is used in the treatment of bromodomain protein 4 diseases by administration through intravenous, oral, sublingual, intramuscular or subcutaneous, or mucocutaneous routes.

[0044] The pharmaceutical composition may be in the form of a liquid or solid preparation, such as a tablet, capsule, or injection. Each dosage form may be prepared using conventional pharmaceutical methods.

[0045] The beneficial effects of the present invention are:

[0046] The present invention, based on a virtual screening method for molecular docking using a water network strategy and biological activity assays, has for the first time discovered 21 compounds with significant inhibitory activity against bromodomain protein 4. These compounds can be used as bromodomain protein 4 inhibitors to treat diseases associated with bromodomain protein 4, providing a new option for current drug research for the treatment of prostate cancer. The present invention's bromodomain protein 4 inhibitors, prepared based on bromodomain protein 4-active compounds with significant inhibitory activity, are highly reliable and easily promoted and popularized for use in the treatment of diseases associated with bromodomain protein 4. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1The water network distribution in the complex of compound 1 and BRD4 protein and its corresponding thermodynamic properties;

[0048] Figure 2 The impact of water networks on screening capacity;

[0049] Figure 3 The results of the BRD4 binding test of compound 1 at a series of concentration gradients are shown;

[0050] Figure 4 The results of the BRD4 binding ability experiments of compounds 2, 9, 10, 11, 14, 15, 16, 17, 20, and 21 at a series of concentration gradients are shown;

[0051] Figure 5 The results of the BRD4 binding ability experiments of compounds 3, 4, and 5 at a series of concentration gradients are shown;

[0052] Figure 6 The results of the BRD4 binding test of compound 6 at a series of concentration gradients are shown;

[0053] Figure 7 The results of the BRD4 binding test of compound 7 at a series of concentration gradients are shown;

[0054] Figure 8 The results of the BRD4 binding test of compound 8 at a series of concentration gradients are shown;

[0055] Figure 9 The results of the BRD4 binding test of compound 12 at a series of concentration gradients are shown;

[0056] Figure 10 The results of the BRD4 binding test of compound 13 at a series of concentration gradients are shown;

[0057] Figure 11 The results of the BRD4 binding test of compound 18 at a series of concentration gradients are shown;

[0058] Figure 12 The results of the BRD4 binding test of compound 19 at a series of concentration gradients are shown;

[0059] Figure 13 The binding conformation of the inhibitor in the BRD4 active pocket (the protein is shown in ribbon shape and the inhibitor is shown in stick shape);

[0060] Figure 14 Schematic diagram of the interaction pattern between the inhibitor and the BRD4 active pocket residues;

[0061] Figure 15The results show the inhibitory ability of compounds 9, 12, 13 and enzalutamide on the proliferation of prostate cancer cells. DETAILED DESCRIPTION

[0062] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0063] The specific embodiments of the present invention are as follows:

[0064] 1) Virtual screening of molecular docking based on water network strategy:

[0065] Experimental principle: Molecular docking method is used to predict, analyze and evaluate the interaction between compounds in the compound database and BRD4, so as to identify inhibitor molecules that can bind to BRD4.

[0066] Experimental method: Based on the crystal structure of BRD4 complex (Protein Structure Database, i.e. RCSB Protein Data Bank, referred to as PDB, number: 3P5O), ​​Schrödinger The Glide module within the molecular simulation software was used to conduct virtual screening studies based on molecular docking. The screening process considered the enhanced effect of the water network on virtual screening capabilities, and the thermodynamic properties of water molecules within the water network were calculated using AMBER software. The compound libraries used for virtual screening included the latest versions of the Chembridge and ChemDiv databases, as well as a three-dimensional structure database of active ingredients in traditional Chinese medicines developed by our research group, which contains over 60,000 compounds. The 2,000 compounds with the highest virtual screening scores were evaluated using the pan-assay interference compounds (PAINS) rule, and molecules containing reactive groups were eliminated.

[0067] Experimental results: First, the thermodynamic properties of each water molecule in the water network were calculated using AMBER software, such as Figure 1 The results show that the binding free energy of each water molecule in the water network is very stable, and the values ​​are all negative and significantly less than 0 (ΔG), and they all occupy their positions well, with an occupancy rate of more than 90%, which also makes them have a significant entropy reduction effect (TΔS) and a smaller exchange number. Although the outer water bridge exchange number is higher, considering that they still have a high occupancy rate of about 60%, the influence of these water molecules cannot be ignored, and they still have good Gibbs free energy (ΔG) and enthalpy change (ΔH). Molecular docking can more accurately determine the small organic molecules that can form strong interactions with BRD4 under the gain effect of the water network, which is shown by Figure 2 It can be seen. Figure 2It is the tested working curve, and the area under the curve is mainly used to evaluate the discrimination ability of the model. That is, the closer the curve is to the upper left corner, the better the sensitivity of the model, and the closer (1-certainty) is to the left, the stronger the discrimination ability of the model. Here, the area under the curve of the screening ability when retaining the water network is 0.77477, which is significantly higher than the area under the curve of 0.65048 when removing water molecules (the area is only 1 at most), which means that the water network has a positive gain effect on virtual screening. Based on the prediction results of molecular docking, more than 200 compounds were purchased from the commercial compound library, and subsequent molecular level binding experiments (EPIgeneous TM Binding Domain Kit A, Cisbio) was used for activity testing, and a number of small molecule compounds with significant BRD4 inhibitory activity were discovered, as shown in Table 1.

[0068] Table 1

[0069]

[0070]

[0071] 2) BRD4 binding assay:

[0072] Experimental Principle: The binding ability of the compound BRD4 was measured using a homogeneous time-resolved fluorescence assay. In this technique, the core energy donor is a trivalent europium ion chelate, which does not emit light on its own and requires excitation with light at both 620nm and 665nm wavelengths in a microplate reader. The 620nm excitation light (donor fluorescence) serves as an internal reference, while the 665nm excitation light (acceptor fluorescence) serves as the reaction indicator. Because the sample interferes with and attenuates both wavelengths, their ratio remains constant, allowing for the measurement of protein-small molecule binding affinity.

[0073] Experimental method: 4 μL BRD4 BD1 protein and 4 μL acetylated lysine histone 4 short peptide ([Lys(5,8,12,16)Ac]H4(1-21)-biotin) were mixed in 10 μL buffer (5 μL streptavidin-acceptor and 5 μL anti-gst-donor), and then 2 μL of 30000nM / 10000nM / 3333.33nM / 1111.11nM / 370.37nM / 123.46nM / 41.15nM / 13.72nM / 4.57nM / 1.52nM (corresponding to experimental wells with different concentration gradients) of test compound (virtual screening compound) was added. If the test compound has a high affinity for BRD4 BD1, it will act as a competitive ligand to replace the native substrate in the binary complex, causing the system's fluorescence polarization value to decrease. If the added test compound has little binding ability to BRD4 BD1, the system's fluorescence polarization value will still remain at a high value. Using a multifunctional microplate reader to measure the change in the system's fluorescence polarization value can quantitatively measure the binding affinity of the virtual screening compound to BRD4.

[0074] Experimental results: The binding ability of different compounds to BRD4 was tested, and it was found that the series of compounds had good binding ability, and their half-inhibitory concentration IC of inhibiting the binding of fluorescent ligands to BRD4 was 50 It is about sub-mole level. Figure 3 、 Figure 4 、 Figure 5 、 Figure 6 、 Figure 7 、 Figure 8 、 Figure 9 、 Figure 10 、 Figure 11 、 Figure 12 and Figure 13 As shown in the figure, the curves of protein activity inhibition rate of these compounds and the logarithm of compound final concentration are given, and IC 50 Table 2 shows the specific values ​​of the experimental results of the 21 compounds of the present invention for their ability to bind to BRD4 under a series of concentration gradients. 50 The values ​​of SQ-9, SQ-12 and SQ-17 are better than those of the lead compounds reported in other literatures, and the IC 50 The values ​​were all below 100 nM, indicating that they bound tightly to the BRD4 protein.

[0075] Table 2

[0076]

[0077]

[0078] 3) Evaluation of the interaction mode between the inhibitor and BRD4:

[0079] Experimental principle: Based on molecular docking and molecular dynamics simulation, the interaction pattern between BRD4 inhibitors and BRD4 is predicted at the atomic scale.

[0080] Experimental steps: Based on the results of molecular docking prediction, a 1000 ns molecular dynamics simulation of the inhibitor / BRD4 was performed using the software AMBER.

[0081] Experimental results: The interaction between the inhibitor and BRD4 obtained by molecular docking prediction and molecular dynamics simulation is as follows Figure 13 and Figure 14 The predicted structure shows that the molecular recognition between the inhibitor and BRD4 is mainly through van der Waals and hydrogen bonding interactions. The hydroxyl group on the dihydroquinoline ring of the inhibitor will interact with asparagine 140 (i.e. Figure 14 Asn 99) forms a stable hydrogen bond, and the dihydroquinoline ring and the phenyl group / cycloheptyl group produce strong van der Waals interactions with multiple hydrophobic residues around them.

[0082] 4) MTT assay to detect the anti-proliferation activity of the compounds against prostate tumor cells:

[0083] Experimental Principle: Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) to water-insoluble blue-purple crystalline formazan, which is deposited in the cells. Dead cells do not have this function. Buffer is added to dissolve the formazan formed in the cells, and its light absorbance is measured at a wavelength of 490nm using an enzyme-linked immunosorbent assay, which can indirectly reflect the number of living cells.

[0084] Experimental steps: Cancer cells were inoculated into 96-well plates at a density of 3000 cells / well using complete medium without androgen. After the cells were stably attached to the wall, 1nM androgen dihydrotestosterone (DHT) and 100000nM / 50000nM / 25000nM / 12500nM / 6250nM / 3125nM / 1562.5nM / 781.25nM / 390.625nM / 195 were simultaneously administered. .3125nM / 97.65625nM / 48.828125nM / 24.4140625nM / 12.20703125nM / 6.103515625nM / 3.051757813nM (corresponding to experimental wells with different concentration gradients) of test compounds (virtual screening compounds, marketed drugs, or dimethyl sulfoxide (DMSO)). After incubation for 4 days, 10 μL of 5 mg / ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was added to each well and incubated in an incubator for another 3 hours. Then, 100 μL of SDS-HCl-PBS triple buffer was added to each well. After incubation at 37°C overnight, the absorbance of each well at 570 nm was measured on a microplate reader and converted into survival rate to obtain the IC value of the administered compound. 50 value.

[0085] Experimental results: Figure 15 The results showed that the compounds of the present invention have a significant ability to inhibit the proliferation of prostate cancer cells LNCaP. With the increase of the final concentration of the compounds, the cell survival rate decreased significantly. Finally, their IC 50 Both can reach levels similar to enzalutamide, a marketed drug for prostate cancer.

[0086] The bromodomain protein 4 active compound of the present invention can be used to prepare drugs that inhibit the activity of bromodomain protein 4, and can be used to prepare pharmaceutically acceptable salts. In addition, the bromodomain protein 4 active compound and the pharmaceutically acceptable salts prepared therefrom can also be used to prepare bromodomain protein 4 inhibitors and the pharmaceutically acceptable salts prepared therefrom, as well as pharmaceutical compositions for preparing prostate cancer cell proliferation inhibitors, anti-prostate tumor drugs and treatment of bromodomain protein 4-related diseases. The bromodomain protein 4 inhibitor prepared based on the bromodomain protein 4 active compound with obvious inhibitory activity of the present invention is applied to the treatment of diseases related to bromodomain protein 4, has high reliability, and is easy to promote and popularize. This application demonstrates the potential advantages of using bromodomain protein 4 active compounds as drugs or pharmaceutical compositions in the field of disease treatment, and opens up new avenues for the treatment of related diseases.

Claims

1. Use of a bromodomain protein 4 active compound based on a water network strategy in the preparation of an anti-prostate tumor drug that inhibits bromodomain protein 4 activity; The structural formula of the bromodomain protein 4 active compound is specifically one of the following structural formulas: Structural formula (1) is N-(3-chlorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (2) is N-(4-fluorophenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide; Structural formula (3) is N-(2-bromo-4-methylphenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide; Structural formula (4) is N-(3-fluoro-4-methylphenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide; Structural formula (5) is N-(2,4-difluorophenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide; Structural formula (6) is N-(4-bromo-2-methylphenyl)-2-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)acetamide; Structural formula (7) is N-(4-fluorobenzyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (8) is N-(2-chloro-4-fluorobenzyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (9) is N-(4-fluorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (10) is 4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)-N-(3-(trifluoromethyl)phenyl)butanamide; Structural formula (11) is N-(2-bromo-4-methylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (12) is N-(2,4-difluorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (13) is N-(5-chloro-2-methoxyphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (14) is N-(2-fluorophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (15) is N-(3-methoxyphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (16) is N-(5-chloro-2,4-dimethoxyphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (17) is N-cycloheptyl-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (18) is N-(4-ethylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (19) is N-(4-isopropylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (20) is N-(4-chloro-2-methoxy-5-methylphenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide; Structural formula (21) is N-(3-cyanophenyl)-4-((1-methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)butanamide.

2. Use of the bromodomain protein 4 active compound based on the water network strategy according to claim 1 in the preparation of pharmaceutically acceptable salts for treating prostate tumors.

3. The use of the bromodomain protein 4 active compound based on the water network strategy according to claim 2 in the preparation of a pharmaceutically acceptable salt for treating prostate tumors, characterized in that: The pharmaceutically acceptable salt for resisting prostate tumors is hydrochloride, phosphate, sulfate, acetate, maleate, citrate, benzenesulfonate, toluenesulfonate, fumarate or tartrate.

4. Use of the bromodomain protein 4 active compound based on the water network strategy according to claim 1 and the pharmaceutically acceptable salt prepared therefrom in the preparation of a bromodomain protein 4 inhibitor, wherein the bromodomain protein 4 inhibitor is used for treating prostate tumors.

5. Use of the bromodomain protein 4 active compound based on the water network strategy according to claim 1 and the pharmaceutically acceptable salt prepared therefrom in the preparation of a prostate cancer cell proliferation inhibitor.

6. Use of the bromodomain protein 4 active compound based on the water network strategy according to claim 1 and the pharmaceutically acceptable salt prepared therefrom in the preparation of anti-prostate tumor drugs.

7. Use of the bromodomain protein 4 active compound based on the water network strategy and the pharmaceutically acceptable salt prepared therefrom according to claim 1 in the preparation of a therapeutic pharmaceutical composition for bromodomain protein 4 diseases, wherein the therapeutic pharmaceutical composition is used for treating prostate tumors.

8. Use of the bromodomain protein 4 active compound based on the water network strategy and the pharmaceutically acceptable salt prepared therefrom according to claim 7 in the preparation of a pharmaceutical composition for treating bromodomain protein 4 diseases, characterized in that: The therapeutic pharmaceutical composition specifically includes an excipient or a diluent.

9. Use of the bromodomain protein 4 active compound based on the water network strategy and the pharmaceutically acceptable salt prepared therefrom according to claim 7 in the preparation of a pharmaceutical composition for treating bromodomain protein 4 diseases, characterized in that: The therapeutic pharmaceutical composition is used for administration via intravenous, oral, sublingual, intramuscular or subcutaneous, or mucocutaneous routes in the treatment of bromodomain protein 4 diseases.

10. Use of the bromodomain protein 4 active compound based on the water network strategy and the pharmaceutically acceptable salt prepared therefrom according to claim 7 in the preparation of a pharmaceutical composition for treating bromodomain protein 4 diseases, characterized in that: The pharmaceutical composition is in the form of a liquid preparation or a solid preparation.

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