Substituted tetrahydroquinoline derivatives and their applications

Abstract

This invention provides a substituted tetrahydroquinoline derivative and its application, which is a class of compounds with AR antagonistic activity. These compounds competitively bind to androgens, blocking AR activation and inhibiting the AR signaling pathway. Experiments have confirmed that the synthesized compounds all possess excellent AR antagonistic activity, showing strong inhibitory activity in the human prostate cancer cell line LNCaP. Therefore, they can be used to prepare drugs for treating diseases with abnormal androgen receptor expression, such as tumors including but not limited to prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, and ovarian cancer. The general structural formula of the compounds is shown in formula (a):

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to substituted tetrahydroquinoline derivatives and their applications. It is a pharmaceutical use of a pharmaceutical composition containing substituted tetrahydroquinoline derivatives, particularly in the preparation of drugs for treating prostate cancer. Background Technology

[0002] Targeted therapy is an important option for patients with advanced prostate cancer or metastatic prostate cancer. The androgen receptor (AR) is one of the key therapeutic targets. AR is a ligand-dependent transcription factor, belonging to the nuclear receptor superfamily. The gene encoding AR is located on human chromosome X, q11-12, and consists of eight exons, encoding a 110 kDa protein of approximately 919 amino acids. AR is mainly composed of four key domains: the N-terminal domain, the DNA-binding domain, the hinge region, and the C-terminal domain. The C-terminal domain contains a ligand-binding pocket (LBP), which is the binding site for androgens. Currently, clinically used AR antagonists target the endogenous ligand-binding pocket of the ARLBP, competitively binding to androgens to block AR activation and inhibit the AR signaling pathway. AR antagonists can be chemically classified into steroidal and non-steroidal types. Due to the hepatotoxic and cardiovascular side effects of steroidal antagonists, and their interference with libido, their clinical efficacy is severely limited. Since their introduction, nonsteroidal anti-inflammatory drugs (NSAIDs), especially second-generation NSAIDs such as enzalutamide and apalutamide, have achieved significant clinical benefits and a strong reputation. However, the development of inherent or acquired resistance, coupled with the excessive similarity in chemical structures of marketed drugs, has led to the simultaneous resistance of multiple NSAIDs. Furthermore, the scarcity of alternative drugs severely restricts the clinical application and selection of these NSAIDs.

[0003] Therefore, designing novel androgen receptor antagonists will help increase drug options for clinical patients and is of great significance for prolonging patient survival. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the technical problem to be solved by the present invention is to provide a substituted tetrahydroquinoline derivative, which is a novel compound with androgen receptor antagonistic activity, and apply it to the preparation of drugs for androgen receptor-mediated diseases such as prostate cancer.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] Previously, patent CN111170943A disclosed a series of benzo[f]cyclopentano[c]quinoline compounds obtained through computer-aided virtual screening and preliminary molecular structure modification, and then a lead compound with androgen receptor antagonistic activity was obtained through biological screening.

[0007] Based on the lead compounds disclosed in the aforementioned patents, this invention modifies their molecular structures and analyzes their structure-activity relationships to obtain a series of novel substituted tetrahydroquinoline derivatives. These compounds exhibit superior androgen receptor antagonistic activity and pharmaceutical properties, providing a potential treatment option for androgen receptor-mediated diseases such as prostate cancer.

[0008] The substituted tetrahydroquinoline derivatives have the general formula (a):

[0009]

[0010] Or its pharmaceutically acceptable salt, stereoisomer, or solvate.

[0011] For the general structural formula (a):

[0012] R1 is selected from C 1-8 Alkyl, C 1-8 Fluoroalkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl, C 3-8 Unsaturated alicyclic group, C 3-8 Saturated alicyclic group, -(CH2) n NR4R5, -(CH2) n OR5, where n is an integer from 1 to 6; R1 or an aromatic ring selected from R6 substituted, including but not limited to benzene ring, furan ring, thiophene ring, pyrrole ring, thiazole ring, pyrazole ring, pyridine ring, pyrimidine ring, pyridazine ring, pyrazine ring, purine ring, etc.

[0013] R2 and R3 are each independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Fluoroalkyl, C 1-6 Alkoxy, fluorinated alkoxy, C 1-6 Halogenated alkyl, halogen, amino, cyano, nitro, hydroxyl, amide, sulfonamide, aldehyde, carboxyl, ester; or

[0014] R2 and R3 combine to form R7-substituted 3-8 membered saturated or unsaturated carbocyclic or heterocyclic rings, including but not limited to benzene rings, thiazole rings, thiophene rings, pyrrole rings, pyridine rings, pyrimidine rings, pyridazine rings, pyrazine rings, etc.; or

[0015] R4 and R5 are independent of each other and can be the same or different. They are selected from hydrogen and C. 1-4 Alkyl, C1-4 Cycloalkyl groups, or NR4R5, are 4-8 cyclic amines, including but not limited to morpholine, piperazine, pyrrolidine, piperidine, etc.

[0016] R6 is selected from hydrogen, halogen, amino, cyano, nitro, hydroxyl, amide, sulfonamide, aldehyde, carboxyl, ester, and C. 1-6 Alkyl, fluorinated alkyl, C 1-6 Alkoxy, fluorinated alkoxy, C 1-6 Haloalkyl, C 2-6 Unsaturated aliphatic hydrocarbon group, C 1-6 Alkylamine group, C 1-6 Dialkylamine group;

[0017] R7 is selected from hydrogen, halogen, amino, cyano, nitro, hydroxyl, amide, sulfonamide, aldehyde, carboxyl, ester, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl groups;

[0018] In ring A, X is selected from O and CH2O.

[0019] Furthermore, in the preferred substituted tetrahydroquinoline compounds of the present invention, R1 in the general formula (a) is selected from, but not limited to, the following groups:

[0020]

[0021] Furthermore, the preferred substituted tetrahydroquinoline compounds of the present invention, in general formula (a), have R2 and R3 combined to form a saturated or unsaturated carbocyclic or heterocyclic ring, preferably from, but not limited to, the following groups:

[0022]

[0023] Alternatively, R2 may be independently selected from hydrogen, and R3 may be selected from, but is not limited to, the following groups: hydrogen, cyano, fluorine, chlorine, methyl, methoxy, trifluoromethyl.

[0024] Furthermore, in the preferred substituted tetrahydroquinoline compounds of the present invention, in the general formula (a), R4 is preferably derived from H;

[0025] Furthermore, in the preferred substituted tetrahydroquinoline compounds of the present invention, in the general formula (a), X in ring A is selected from: O, CH2O.

[0026] It should be understood that the present invention includes all combinations and subgroups of specific groups as defined in the present invention, including substituents as defined in the brief description above and exemplified in the various embodiments throughout the specification.

[0027] More specifically, the preferred structural formulas of the substituted tetrahydroquinoline derivatives of the present invention with general formula (a) are shown in Table 1.

[0028] Table 1

[0029]

[0030]

[0031] The compounds described above can be prepared by the following steps, but are not limited to these methods.

[0032] Synthetic methods for compounds (including analogs) of general formula (a):

[0033]

[0034] The specific reaction process can be:

[0035] Substituted aldehydes (compound A) and substituted anilines (compound B) dissolved in solvents undergo a Povarov reaction with the corresponding electron-rich olefins (compound C) under Lewis acid catalysis. Separation by column chromatography yields substituted tetrahydroquinoline compound D. Solvents used include acetonitrile, tetrahydrofuran, toluene, dichloromethane, trichloromethane, N,N-dimethylformamide, N,N-dimethylacetamide, dioxane, and methanol. Lewis acids include trifluoroacetic acid, indium trichloride, aluminum trichloride, p-toluenesulfonic acid, bismuth trichloride, and ytterbium trifluoromethanesulfonate.

[0036] The compounds of general formula (a) provided by this invention, or their pharmaceutically acceptable salts, can target and antagonize AR, exhibiting significant AR transcriptional repression. In particular, compound II-1 can effectively inhibit the expression level of eGFP protein in the prostate cancer cell line LNCaP.

[0037] A second object of the present invention is to provide a pharmaceutical composition comprising at least one active ingredient (a substituted tetrahydroquinoline compound) as described above and at least one pharmaceutically acceptable carrier or excipient for use in the preparation of an androgen receptor (AR) antagonist.

[0038] The term "pharmaceutically acceptable carrier" refers to conventional drug carriers in the pharmaceutical field, including conventional diluents and excipients such as water, fillers such as starch, binders such as cellulose derivatives and gelatin, humectants such as glycerin, disintegrants such as agar and calcium carbonate, absorption enhancers such as quaternary ammonium compounds, surfactants such as hexadecyl alcohol, adsorbents such as kaolin and soap clay, lubricants such as talc, and flavoring agents and sweeteners may be added when necessary.

[0039] Pharmaceutical formulations are suitable for administration via any appropriate route, such as oral (including sublingual or sublingual), rectal, nasal, topical (including sublingual, sublingual, or transdermal), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, or intradermal injection). These formulations can be prepared by any method known in the field of pharmaceutics, such as by mixing the active ingredient with a carrier or excipient.

[0040] In this invention, "pharmaceutically acceptable salt" refers to salts prepared by conventional methods, including but not limited to organic acid salts, inorganic acid salts, organic base salts, and inorganic base salts. The organic acid salts include, but are not limited to, oxalates, lactates, p-toluenesulfonates, maates, citrates, fumarates, camphorsulfonates, and methanesulfonates; the inorganic acid salts include, but are not limited to, nitrates, sulfates, hydrohalates, and phosphates. The organic base salts include, but are not limited to, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, triethylamine, and tert-butylamine. The inorganic base salts include, but are not limited to, sodium hydroxide, potassium hydroxide, and lithium hydroxide.

[0041] A third objective of this invention is to provide a compound or pharmaceutical composition as described above for use in the preparation of a medicament for treating diseases related to abnormal expression or mutation of the androgen receptor. Experiments have confirmed that the synthesized compounds all possess excellent AR antagonistic activity and exhibit strong inhibitory activity in the human prostate cancer cell line LNCaP.

[0042] Preferably, the drug is used to prevent or treat diseases such as tumors caused by abnormal expression or mutation of androgen receptors.

[0043] More preferably, the tumor is selected from solid tumors, including prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, metastatic castration-resistant prostate cancer, breast cancer, ovarian cancer, etc.

[0044] The therapeutic mechanism of the drug includes: its active ingredient, replacing tetrahydroquinoline derivatives, inhibits tumor cell proliferation by antagonizing androgen receptor activity. Furthermore, the substituted tetrahydroquinoline derivatives competitively bind to the endogenous ligand-binding pocket (LBP) of the AR receptor, thereby exerting an antagonistic effect on androgen receptors.

[0045] Antitumor drugs that can be used in combination with the compounds provided by this invention or their pharmaceutically acceptable salts include, but are not limited to, mitotic inhibitors (such as vincristine, vindesine, etc.); microtubule degradation inhibitors (such as paclitaxel); antimetabolites (such as 5-fluorouracil, methotrexate, and cytarabine, etc.); alkylating agents (such as cisplatin, carboplatin, and cyclophosphamide, etc.); intercalable antibiotics (such as areopyram, mitomycin, and bleomycin, etc.); enzymes (such as aspartase, etc.); and topoisomerase inhibitors (such as etoposide and camptothecin, etc.).

[0046] The beneficial effects of this invention are as follows:

[0047] This invention provides a novel class of substituted tetrahydroquinoline compounds with androgen receptor antagonistic activity. One of the key target sites of these compounds is the androgen receptor ligand-binding pocket, enabling them to competitively bind to androgens, blocking AR activation and inhibiting the AR signaling pathway. The compounds provided by this invention exhibit high antagonistic activity against androgen receptor transcription, demonstrating good biological activity at both the molecular and cellular levels, and possessing good safety. Therefore, they can be applied to the preparation of drugs for treating diseases with abnormal androgen receptor expression, such as tumors including but not limited to prostate cancer, metastatic prostate cancer, castration-resistant prostate cancer, breast cancer, and ovarian cancer. Attached Figure Description

[0048] Figure 1 The representative compounds I-1, II-1, III-3, V-1, V-5, V-6 and V-17 were shown to inhibit the proliferation of LNCaP cells.

[0049] Figure 2 The representative compounds I-1, II-1, III-3, V-1, V-5, V-6 and V-17 were shown to inhibit the proliferation of DU145 cells.

[0050] Figure 3 The representative compounds I-1, II-1, III-3, V-1, V-5, V-6 and V-17 were shown to inhibit the proliferation of PC3 cells.

[0051] Figure 4 Representative compounds I-1, II-1, III-3, and V-1 in AR LBD F877L / T878A Transcriptional repression activity in a double-mutant enzalutamide resistance model.

[0052] Figure 5 The effects of compounds II-1 and V-1 on exocrine PSA levels are represented.

[0053] Figure 6 The figure shows the experimental results for target verification of compounds II-1 and V-1.

[0054] Figure 7 This is the X-ray diffraction pattern of compound Ⅲ-2 in a single crystal. Detailed Implementation

[0055] The present invention will be further described in conjunction with the accompanying drawings and embodiments. The specific embodiments included below are for illustrative purposes only and should not be construed as limiting the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0056] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0057] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0058] Example 1: Preparation of target molecules I-1 to I-2

[0059] Preparation of (3aR,4R,11cR)-4-(4-chlorophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound I-1)

[0060]

[0061] 4-Chlorobenzaldehyde 1 (1.0 mmol), 2-naphthylamine 2 (1.0 mmol), indium trichloride InCl3 (0.1 mmol), and anhydrous acetonitrile (5 mL) were sequentially added to a reaction flask. After stirring at room temperature for 10 min, 2,3-dihydrofuran 3 (1.2 mmol) was added, and the reaction was continued with stirring at room temperature. After the reaction was monitored by TLC to indicate completion, the solvent was removed under reduced pressure, water (30 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined and washed with saturated NaCl solution (30 mL), dried over anhydrous Na2SO4, concentrated under reduced pressure, and the crude product was separated by column chromatography (ethyl acetate / petroleum ether = 1:10) to obtain a white solid.

[0062] Yield 41%; ESI-MS: 336.1 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.97(d,J=7.0Hz,1H),7.68(d,J=7.0Hz,1H),7.59(d,J=8.5Hz, 1H),7.55(d,J=8.5Hz,2H),7.47(d,J=8.5Hz,2H),7.40-7.37(m,1H),7.21-7.18(m,1H), 7.05(d,J=9.0Hz,1H),6.25(s,1H),5.68(d,J=8.0Hz,1H),4.62(d,J=3.0Hz,1H),3.62-3 .57(m,1H),3.53-3.49(m,1H),2.93-2.87(m,1H),2.07-1.99(m,1H),1.46-1.39(m,1H).

[0063] Preparation of (4aR,5R,12cR)-5-(4-chlorophenyl)-2,3,4a,5,6,12c-hexahydro-1H-benzo[f]pyrano[2,3-c]quinoline (compound I-2)

[0064] Compound I-2 was prepared following the synthesis of I-1, except that 2,3-dihydrofuran was replaced with 3,4-dihydro-2H-pyran, yielding a white solid. Yield: 35%; ESI-MS: 350.1 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.10(d,J=8.5Hz,1H),7.66(d,J=7.0Hz,1H),7.59(d,J=8.5Hz,1H ),7.47(d,J=8.5Hz,2H),7.39(d,J=8.5Hz,2H),7.36-7.33(m,1H),7.16-7.13(m,1H),7.0 0(d,J=8.5Hz,1H),6.53(s,1H),5.40(s,1H),3.49-3.45(m,1H),3.14-3.13(m,1H),2.30- 2.27(m,1H),1.77-1.75(m,1H),1.51-1.50(m,1H),1.30-1.26(m,1H),1.02-1.01(m,1H).

[0065] Example 2: Preparation of target molecules II-1 to II-13

[0066] Preparation of (3aR,4R,11cR)-4-(4-chlorophenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound II-1)

[0067] Compound II-1 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 6-aminoquinoline, yielding a white solid. Yield: 50%; ESI-MS: 337.1 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ8.57 (dd, J1=4.0Hz, J2=1.5Hz, 1H), 8.35-8.31 (m, 1H), 7.70 (d, J=9.0Hz,1H),7.60-7.53(m,2H),7.51-7.44(m,2H),7.38(dd,J1=8.5Hz,J2=4.0Hz,1H), 7.28(d,J=9.0Hz,1H),6.50(s,1H),5.68(d,J=8.0Hz,1H),4.68(d,J=3.0Hz,1H),3.67-3 .57(m,1H),3.57-3.48(m,1H),2.95-2.84(m,1H),2.09-1.94(m,1H),1.49-1.39(m,1H).

[0068] Preparation of (8aR,8R,11cR)-8-(4-chlorophenyl)-7,8,8a,9,10,11a-hexahydrofuran[2,3':4,5]pyrido[3,2-f]quinoxaline (compound II-2)

[0069] Compound II-2 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 6-aminoquinoxaline, yielding a white solid. Yield: 46%; ESI-MS: 338.0 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.74(d,J=2.0Hz,1H),8.56(d,J=2.0Hz,1H),7.76(d,J =9.0Hz,1H),7.54(d,J=8.5Hz,2H),7.46(d,J=8.5Hz,2H),7.40(d,J=9.0Hz,1H) ,6.88(s,1H),5.84(d,J=8.5Hz,1H),4.65(d,J=4.0Hz,1H),3.50(q,J=8.5Hz,1 H),3.42-4.37(m,1H),2.99-2.90(m,2H),2.06-1.92(m,1H),1.53-1.40(m,2H).

[0070] Preparation of (3aR,4R,11cR)-4-(4-chlorophenyl)-2,3,3a,4,5,11c-hexahydrofurano[2',3':4,5]pyrido[3,2-f]quinazoline (compound II-3)

[0071] Compound II-3 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 6-aminoquinazoline, yielding a white solid. Yield: 48%; ESI-MS: 338.8 [M+H] + .

[0072] Preparation of (3aR,4R,11cR)-4-(4-chlorophenyl)-2,3,3a,4,5,11c-hexahydrofuran[3,2-k][3,7]phenanthroline (compound II-4)

[0073] Compound II-4 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 6-aminophenanthroline, yielding a white solid. Yield 49%; ESI-MS: 337.8 [M+H] + .

[0074] Preparation of (3aR,4R,10cR)-4-(4-chlorophenyl)-2,3,3a,4,5,10c-hexahydrofurano[3,2-c]thieno[3,2-f]quinoline (compound II-5)

[0075] Compound II-5 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 5-aminobenzothiophene, yielding a yellow solid. Yield: 37%; ESI-MS: 342.1 [M+H] + .

[0076] Preparation of (3aR,4R,10cR)-4-(4-chlorophenyl)-2,3,3a,4,5,10c-hexahydrofurano[3,2-c]pyrrolo[3,2-f]quinoline (compound II-6)

[0077] Compound II-6 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 5-aminobenzopyrrole, yielding a white solid. Yield: 40%; ESI-MS: 325.1 [M+H] + .

[0078] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound II-7)

[0079] Compound II-7 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with aniline, yielding a white solid. Yield: 38%; ESI-MS: 286.7 [M+H] + ; 1H NMR(500MHz,DMSO-d6)δ7.54-7.47(m,2H),7.47-7.40(m,2H),7.17-7.09(m,1H),7.03-6.91(m,1H),6.73-6.67(m,1H),6.67-6.58(m,1H),5.91( s,1H),5.13(d,J=8.5Hz,1H),4.62(d,J=3.0Hz,1H),3.57(dd,J1=8.5Hz, J2=5.0Hz,2H),2.72-2.62(m,1H),2.01-1.84(m,1H),1.39-1.28(m,1H).

[0080] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-8-methyl-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound II-8)

[0081] Compound II-8 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with p-methylaniline, yielding a white solid. Yield: 44%; ESI-MS: 300.7 [M+H] + .

[0082] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline-8-nitrile (compound II-9)

[0083] Compound II-9 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 4-cyanoaniline, yielding a white solid. Yield: 50%; ESI-MS: 311.7 [M+H] + .

[0084] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-8-fluoro-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound II-10)

[0085] Compound II-10 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 4-fluoroaniline, yielding a white solid. Yield: 43%; ESI-MS: 304.1 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ7.50(d,J=8.5Hz,2H),7.44(d,J=8.5Hz,2H),6.90(dd,J1=9.5Hz,J2=3.0Hz,1H),6.86(td,J1=8.5Hz,J2=3.0Hz,1H),6.71(dd, J1=8.5Hz, J2=5.0Hz,1H),5.11(d,J=8.0Hz,1H),4.59(s,1H),3.64-3.56(m ,1H),2.67(m,1H),2.53-2.51(m,1H),1.97-1.81(m,1H),1.36-1.31(m,1H).

[0086] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-8-chloro-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound II-11)

[0087] Compound II-11 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 4-chloroaniline, yielding a white solid. Yield: 45%; ESI-MS: 321.2 [M+H] + ; 1 H NMR (500MHz, DMSO-d6)7.49(d,J=8.5Hz,2H),7.47-7.42(m,2H),7.10(d,J=2.5Hz,1H),7.02(dd,J1=8.5Hz,J2=2.5Hz,1H),6.71(d,J=8.5Hz,1H ),5.11(d,J=8.0Hz,1H),4.63(d,J=3.5Hz,1H),3.62-3.56(m,2H),2.72 -2.61(m,1H),2.53-2.51(m,1H),1.93-1.81(m,1H),1.38-1.34(m,1H).

[0088] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-8-(trifluoromethyl)-2,3,3a,4,5,9b-hexahydrofluoro[3,2-c]quinoline (compound II-12)

[0089] Compound II-12 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 4-trifluoromethylaniline, yielding a white solid. Yield: 37%; ESI-MS: 354.7 [M+H] + .

[0090] Preparation of (3aR,4R,9cR)-4-(4-chlorophenyl)-8-(methoxy)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound II-13)

[0091] Compound II-13 was prepared following the synthesis of I-1, except that 2-naphthylamine was replaced with 4-methoxyaniline, yielding a white solid. Yield: 43%; ESI-MS: 316.8 [M+H] + .

[0092] Example 3: Preparation of target molecules III-1 to III-11

[0093] Preparation of (3aR,4R,11cR)-4-(4-fluorophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-1)

[0094] Compound III-1 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde, yielding a white solid. Yield: 45%; ESI-MS: 320.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ7.97 (dd, J1=8.5Hz, J2=1.5Hz, 1H), 7.68 (dd, J1=8.5Hz, J2=1.5H z,1H),7.62-7.54(m,3H),7.42-7.35(m,1H),7.27-7.21(m,2H),7.23-7.16(m,1H),7.04 (d,J=8.5Hz,1H),6.29-6.19(m,1H),5.68(d,J=8.0Hz,1H),4.63(d,J=3.5Hz,1H),3.67- 3.56(m,1H),3.56-3.47(m,1H),2.95-2.85(m,1H),2.13-1.99(m,1H),1.50-1.38(m,1H).

[0095] Preparation of (3aR,4R,11cR)-4-(4-bromophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-2)

[0096] Compound III-2 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-bromobenzaldehyde, yielding a white solid. Yield: 45%; ESI-MS: 381.2 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ7.96 (dd, J1=8.5Hz, J2=1.0Hz, 1H), 7.68 (dd, J1=8.5Hz, J2=1.0Hz, 1H), 7.63-7.56(m,3H),7.54-7.46(m,2H),7.42-7.35(m,1H),7.24-7.16(m,1H),7.04(d,J=8.5Hz,1 H),6.25(s,1H),5.68(d,J=8.0Hz,1H),4.61(d,J=3.0Hz,1H),3.60(td,J1=8.5Hz,J2=6.5Hz,1H ),3.51(td,J1=8.0Hz,J2=3.5Hz,1H),2.96-2.86(m,1H),2.09-1.96(m,1H),1.48-1.38(m,1H).

[0097] Preparation of (3aR,4R,11cR)-4-(4-cyanophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-3)

[0098] Compound III-3 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-cyanobenzaldehyde, yielding a white solid. Yield: 37%; ESI-MS: 327.3 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.97(d,J=8.5Hz,1H),7.90-7.84(m,2H),7.73(d,J=8.0Hz,2H),7.69( dd, J1=8.0Hz, J2=1.5Hz,1H),7.61(d,J=8.5Hz,1H),7.43-7.35(m,1H),7.24-7.17(m,1H),7.0 5(d,J=8.5Hz,1H),6.36(s,1H),5.68(d,J=8.0Hz,1H),4.72(d,J=3.5Hz,1H),3.64-3.55(m,1H ),3.49(td,J1=8.5Hz,J2=3.5Hz,1H),3.00-2.91(m,1H),2.07-1.94(m,1H),1.46-1.34(m,1H).

[0099] Preparation of (3aR,4R,11cR)-4-(3-fluorophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-4)

[0100] Compound III-4 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 3-fluorobenzaldehyde, yielding a white solid. Yield: 48%; ESI-MS: 320.3 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.97(d,J=8.5Hz,1H),7.68(dd,J1=8.0Hz,J2=1.5Hz,1H),7.59(d,J=8.5Hz, 1H),7.49-7.40(m,1H),7.41-7.33(m,3H),7.24-7.16(m,1H),7.18-7.10(m,1H),7.06(d,J=8.5Hz,1 H),6.27(d,J=1.5Hz,1H),5.68(d,J=8.0Hz,1H),4.65(d,J=3.0Hz,1H),3.60(td,J1=8.5Hz,J2=6.5H z,1H),3.52(td,J1=8.0Hz,J2=3.0Hz,1H),3.00-2.86(m,1H),2.11-1.97(m,1H),1.48-1.38(m,1H).

[0101] Preparation of (3aR,4R,11cR)-4-(3-chlorophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-5)

[0102] Compound III-5 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 3-chlorobenzaldehyde, yielding a white solid. Yield: 48%; ESI-MS: 336.8 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ7.97 (dd, J1=8.5Hz, J2=1.5Hz, 1H), 7.68 (dd, J1=8.0Hz, J2=1.5Hz, 1H), 7.63-7 .57(m,2H),7.52-7.46(m,1H),7.44(t,J=8.0Hz,1H),7.42-7.35(m,2H),7.24-7.16(m,1H),7.06(d,J=8 .5Hz,1H),6.28(d,J=1.5Hz,1H),5.68(d,J=8.0Hz,1H),4.64(d,J=3.0Hz,1H),3.60(td,J1=8.5Hz,J2=6 .5Hz,1H),3.52(td,J1=8.0Hz,J2=3.5Hz,1H),2.99-2.89(m,1H),2.11-1.97(m,1H),1.49-1.37(m,1H).

[0103] Preparation of (3aR,4R,11cR)-4-(2-fluorophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-6)

[0104] Compound III-6 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 2-fluorobenzaldehyde, yielding a white solid. Yield: 48%; ESI-MS: 320.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ7.97 (dd, J1=8.5Hz, J2=1.0Hz, 1H), 7.74 (td, J1=8.0Hz, J2=1.5Hz, 1H), 7.69 (dd, J1=8.0Hz, J2=1.5Hz,1H),7.60(d,J=8.5Hz,1H),7.45-7.34(m,2H),7.30-7.25(m,1H),7.25-7.18(m,2H) ,7.05(d,J=8.5Hz,1H),6.20(s,1H),5.70(d,J=8.0Hz,1H),4.88(d,J=3.0Hz,1H),3.61(td,J1=8.5Hz,J2 =6.5Hz,1H),3.52(td,J1=8.5Hz,J2=3.5Hz,1H),3.08-2.89(m,1H),2.16-2.00(m,1H),1.55-1.37(m,1H).

[0105] Preparation of (3aR,4R,11cR)-4-(2-chlorophenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-7)

[0106] Compound III-7 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 2-chlorobenzaldehyde, yielding a white solid. Yield: 49%; ESI-MS: 336.8 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ7.99 (d, J=8.5Hz, 1H), 7.82 (dd, J1=8.0Hz, J2=1.5Hz, 1H), 7.68(dd,J1=8.0Hz,J2=1.5Hz,3H),7.50(dd,J1=8.0Hz,J2=1.5Hz,1H),7.25-7.14 (m,3H),7.05(d,J=6.5Hz,1H),6.20(s,1H),5.72(d,J=8.5Hz,1H),4.93(d,J=3.0H z,1H),3.68-3.51(m,2H),3.14-3.00(m,1H),2.48-2.41(m,1H),1.40-1.32(m,1H).

[0107] Preparation of (3aR,4R,11cR)-4-(4-trifluoromethylphenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-8)

[0108] Compound III-8 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-trifluoromethylbenzaldehyde, yielding a white solid. Yield: 46%; ESI-MS: 370.1 [M+H] + .

[0109] Preparation of (3aR,4R,11cR)-4-(4-methoxyphenyl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-9)

[0110] Compound III-9 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-methoxybenzaldehyde, yielding a white solid. Yield: 36%; ESI-MS: 332.4 [M+H] + .

[0111] Preparation of (3aR,4R,11cR)-4-(5-chlorothiophene-2-yl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-10)

[0112] Compound III-10 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 5-chlorothiophene-2-carboxaldehyde, yielding a yellow solid. Yield: 34%; ESI-MS: 342.8 [M+H] + .

[0113] Preparation of (3aR,4R,11cR)-4-(5-chlorofuran-2-yl)-2,3,3a,4,5,11c-hexahydrobenzo[f]furan[3,2-c]quinoline (compound III-11)

[0114] Compound III-11 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 5-chlorofuran-2-carboxaldehyde, yielding a white solid. Yield: 42%; ESI-MS: 326.7 [M+H] + .

[0115] Example 3: Preparation of target molecules IV-1 to IV-7

[0116] Preparation of (8aR,8R,11cR)-8-(4-fluorophenyl)-7,8,8a,9,10,11a-hexahydrofuran[2',3',4,5]pyrido[3,2-f]quinoxaline (compound IV-1)

[0117] Compound IV-1 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoxaline, yielding a yellow solid. Yield: 48%; ESI-MS: 322.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6)δ

[0118] 8.74(d,J=2.0Hz,1H),8.56(d,J=2.0Hz,1H),7.76(d,J=9.0Hz,1H),7.60- 7.50(m,2H),7.40(d,J=9.0Hz,1H),7.29-7.19(m,2H),6.87(s,1H),5.84(d ,J=8.5Hz,1H),4.65(d,J=3.5Hz,1H),3.51(td,J1=8.5Hz,J2=6.5Hz,1H),3 .53-3.48(m,1H),3.01-2.90(m,1H),2.04-1.96(m,1H),1.53-1.42(m,1H).

[0119] Preparation of (3aR,4R,11cR)-8-(4-cyanophenyl)-7,8,8a,9,10,11a-hexahydrofuran[2',3',4,5]pyrido[3,2-f]quinoxaline (compound IV-2)

[0120] Compound IV-2 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-cyanobenzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoxaline, yielding a yellow solid. Yield: 40%; ESI-MS: 329.3 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ8.75(d,J=2.0Hz,1H),8.58(d,J=2.0Hz,1H),7.91-7.85(m,2H),7.78(d,J=9.0Hz,1H),7.71(d,J=8.0Hz,2H),7.40(d,J=9 .0Hz,1H),6.98(s,1H),5.83(d,J=8.0Hz,1H),4.75(d,J=4.0Hz,1H),3.5 5-3.46(m,1H),3.05-2.93(m,1H),2.03-1.91(m,1H),1.54-1.40(m,1H).

[0121] Preparation of (3aR,4R,9cR)-4-(4-fluorophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound IV-3)

[0122] Compound IV-3 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde and 2-naphthylamine was replaced with aniline, yielding a white solid. Yield: 41%; ESI-MS: 270.1 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ7.55-7.47(m,2H),7.25-7.16(m,2H),7.16-7.10(m,1H),7.01-6.94(m,1H),6.73-6.67(m,1H),6.67-6.59(m,1H),5.89( s,1H),5.13(d,J=8.0Hz,1H),4.62(d,J=3.0Hz,1H),3.57(dd,J1=8.5Hz, J2=5.0Hz,2H),2.73-2.58(m,1H),1.99-1.87(m,1H),1.41-1.27(m,1H).

[0123] Preparation of (3aR,4R,9cR)-8-chloro-4-(4-fluorophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound IV-4)

[0124] Compound IV-4 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde and 2-naphthylamine was replaced with 4-chloroaniline, yielding a white solid. Yield: 45%; ESI-MS: 304.1 [M+H] + ; 1 H NMR (500MHz, DMSO-d6)δ

[0125] 7.54-7.47(m,2H),7.25-7.18(m,2H),7.10(d,J=2.5Hz,1H),7.02(dd,J1=8.5Hz,J2=2.5Hz,1H),6.71(d,J=8.5Hz,1H),5.11(d,J= 8.0Hz,1H),4.64(d,J=3.0Hz,1H),3.65-3.54(m,2H),2.72-2.60(m,1H),2.53-2.51(m,1H),1.96-1.84(m,1H),1.40-1.30(m,1H).

[0126] Preparation of (3aR,4R,9cR)-8-chloro-4-(4-cyanophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound IV-5)

[0127] Compound IV-5 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-cyanobenzaldehyde and 2-naphthylamine was replaced with 4-chloroaniline, yielding a white solid. Yield: 38%; ESI-MS: 311.7 [M+H] + .

[0128] Preparation of (3aR,4R,9cR)-8-fluoro-4-(4-fluorophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound IV-6)

[0129] Compound IV-6 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde and 2-naphthylamine was replaced with 4-fluoroaniline, yielding a white solid. Yield: 48%; ESI-MS: 288.3 [M+H] + ; 1 H NMR (500MHz, Chloroform-d)δ

[0130] 7.54-7.48(m,2H),7.25-7.18(m,2H),6.90(dd,J1=9.5Hz,J2=3.0Hz,1H),6.85(td,J1=8.5Hz,J2=3.0Hz,1H),6.70(dd,J1=8.5Hz,J2=5.0H z,1H),5.11(d,J=8.0Hz,1H),4.59(s,1H),3.66-3.53(m,2H),2.73-2.61(m,1H),2.53-2.51(m,1H),2.00-1.83(m,1H),1.42-1.26(m,1H).

[0131] Preparation of (3aR,4R,9cR)-8-cyano-4-(4-fluorophenyl)-2,3,3a,4,5,9b-hexahydrofuran[3,2-c]quinoline (compound IV-7)

[0132] Compound IV-7 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde and 2-naphthylamine was replaced with 4-cyanoaniline, yielding a white solid. Yield: 58%; ESI-MS: 295.1 [M+H] + ; 1 H NMR(500MHz,Chloroform-d)δ7.51(dd,J1=2.0Hz,J2=0.5Hz,1H),7.48(dd,J1=9.0Hz,J2=2.0Hz,1H),7.34-7.27(m,2H),7.05-6.98(m,2H),6.90(d,J=9.0Hz, 1H),4.94(d,J=7.5Hz,1H),4.77-4.75(m,1H),4.54(dd,J1=5.5Hz,J2=4.5Hz,1 H),3.91-3.80(m,2H),3.40-3.34(m,1H),2.42-2.38(m,1H),2.18-2.10(m,1H).

[0133] Example 5: Preparation of target compounds V-1 to V-17

[0134] Preparation of (3aR,4R,11cR)-8-(4-fluorophenyl)-7,8,8a,9,10,11a-hexahydrofuran[2,3':4,5]pyrido[3,2-f]quinoline (compound V-1)

[0135] Compound V-1 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-fluorobenzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoline, yielding a yellow solid. Yield: 48%; ESI-MS: 321.3 [M+H] + ; 1H NMR (500MHz, DMSO-d6) δ8.56 (dd, J1=4.0Hz, J2=1.5Hz, 1H), 8.36-8.30 (m, 1H), 7.70 (d,J=9.0Hz,1H),7.62-7.53(m,2H),7.38(dd,J1=8.5Hz,J2=4.0Hz,1H),7.30-7.21 (m,3H),6.52-6.47(m,1H),5.68(d,J=8.0Hz,1H),4.68(d,J=3.0Hz,1H),3.66-3.57 (m,1H),3.58-3.50(m,1H),2.94-2.85(m,1H),2.10-1.97(m,1H),1.50-1.40(m,1H).

[0136] Preparation of (3aR,4R,11cR)-8-(4-cyanophenyl)-7,8,8a,9,10,11a-hexahydrofuran[2,3':4,5]pyrido[3,2-f]quinoline (compound V-2)

[0137] Compound V-2 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-cyanobenzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoline, yielding a yellow solid. Yield: 38%; ESI-MS: 328.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.58 (dd, J1=4.0Hz, J2=1.5Hz, 1H), 8.39-8.28 (m, 1H), 7.9 6-7.84(m,2H),7.82-7.72(m,3H),7.39(dd,J1=8.5Hz,J2=4.0Hz,1H),7.28(d,J=9 .0Hz,1H),6.59(s,1H),5.69(d,J=8.0Hz,1H),4.78(d,J=3.0Hz,1H),3.66-3.57(m ,1H),3.55-3.47(m,1H),3.01-2.88(m,1H),2.06-1.91(m,1H),1.47-1.34(m,1H).

[0138] Preparation of (3aR,4R,11cR)-4-(4-ethoxyphenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-3)

[0139] Compound V-3 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-ethoxybenzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoline, yielding a yellow solid. Yield: 43%; ESI-MS: 347.4 [M+H]+ ; 1 H NMR(500MHz,Chloroform-d)δ8.68(dd,J1=4.0Hz,J2=1.5Hz,1H),8.51-8.44(m,1H),7.85(d,J=9 .0Hz,1H),7.44-7.39(m,2H),7.37(dd,J1=8.5Hz,J2=4.0Hz,1H),7.05(d,J=9.0Hz,1H),6.97-6.9 0(m,2H),5.78(d,J=8.0Hz,1H),4.66(d,J=3.0Hz,1H),4.11-4.07(m,1H),4.06(q,J=7.0Hz,2H),3 .84-3.72(m,2H),3.01-2.88(m,1H),2.36-2.21(m,1H),1.71-1.60(m,1H),1.44(t,J=7.0Hz,3H).

[0140] Preparation of (3aR,4R,11cR)-4-(4-propoxyphenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-4)

[0141] Compound V-4 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-propoxybenzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoline, yielding a yellow solid. Yield: 52%; ESI-MS: 361.4 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.55 (dd, J1=4.0Hz, J2=1.5Hz, 1H), 8.41-8.27 (m, 1H), 7.68 (d, J=9.0Hz, 1H), 7.4 7-7.40(m,2H),7.37(dd,J1=8.5Hz,J2=4.0Hz,1H),7.28(d,J=9.0Hz,1H),7.01-6.85(m,2H),6.41(d,J=1. 5Hz,1H),5.67(d,J=8.0Hz,1H),4.59(d,J=3.0Hz,1H),3.98-3.89(m,1H),3.65-3.58(m,1H),3.58-3.50( m,1H),2.93-2.79(m,1H),2.09-2.00(m,1H),1.78-1.67(m,2H),1.51-1.42(m,1H),0.99(t,J=7.5Hz,3H).

[0142] Preparation of (3aR,4R,11cR)-4-(4-isopropoxyphenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-5)

[0143] Compound V-5 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by 4-isopropoxybenzaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 43%; ESI-MS: 361.2 [M+H] + ; 1 H NMR (500MHz, Chloroform-d)δ

[0144] 8.68(dd,J1=4.0Hz,J2=1.5Hz,1H),8.52(d,J=8.5Hz,1H),7.91(d,J=9.0Hz,1H),7.3 9(dd,J1=8.5Hz,J2=4.5Hz,3H),7.07(d,J=9.0Hz,1H),6.94-6.88(m,2H),5.78(d,J=8 .0Hz,1H),4.67(d,J=3.0Hz,1H),4.62-4.50(m,1H),4.17-4.10(m,1H),3.84-3.69(m ,2H),3.02-2.90(m,1H),2.38-2.23(m,1H),1.75-1.62(m,1H),1.36(d,J=6.0Hz,6H).

[0145] Preparation of (3aR,4R,11cR)-4-(4-cyclopropoxyphenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-6)

[0146] Compound V-6 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by 4-cyclopropoxybenzaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 42%; ESI-MS: 359.1 [M+H] + ; 1H NMR(500MHz,Chloroform-d)δ8.69(dd,J1=4.0Hz,J2=1.5Hz,1H),8.50(d,J=8.5Hz,1H),7.88(d,J=9.0Hz, 1H),7.42(dd,J1=8.5Hz,J2=2.5Hz,2H),7.38(dd,J1=8.5Hz,J2=4.0Hz,1H),7.12-7.01(m,2H),7.01-6.92 (m,1H),6.16-6.02(m,1H),5.79(dd,J1=8.0Hz,J2=2.0Hz,1H),5.50-5.40(m,1H),5.34-5.30(m,1H),4.67 (d,J=3.0Hz,1H),4.60-4.54(m,1H),4.11(s,1H),3.85-3.71(m,3H),3.03-2.88(m,1H),2.40-2.23(m,1H).

[0147] Preparation of (3aR,4R,11cR)-4-(4-(trifluoromethoxy)phenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-7)

[0148] Compound V-7 was prepared following the synthesis of I-1, except that 4-chlorobenzaldehyde was replaced with 4-(trifluoromethoxy)benzaldehyde and 2-naphthylamine was replaced with 6-aminoquinoline, yielding a yellow solid. Yield 40%; ESI-MS: 387.1 [M+H] + ; 1 H NMR(500MHz,Chloroform-d)δ8.69(dd,J1=4.0Hz,J2=1.5Hz,1H),8.57-8.47(m,1H),7. 91(d,J=9.0Hz,1H),7.59-7.51(m,2H),7.40(dd,J1=8.5Hz,J2=4.0Hz,1H),7.29-7.27( m,1H),7.08(d,J=9.0Hz,1H),5.80(d,J=8.0Hz,1H),4.75(d,J=3.0Hz,1H),4.14(d,J=1 1.0Hz,1H),3.82-3.73(m,2H),3.05-2.92(m,1H),2.34-2.20(m,1H),1.68-1.58(m,1H).

[0149] Preparation of (3aR,4R,11cR)-4-(4-(difluoromethoxy)phenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-8)

[0150] Compound V-8 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by 4-(difluoromethoxy)benzaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 50%; ESI-MS: 369.1 [M+H] + ; 1 H NMR(500MHz,Chloroform-d)δ8.69(dd,J1=4.0Hz,J2=1.5Hz,1H),8.57-8.44(m,1H),7.93 (d,J=9.0Hz,1H),7.59-7.47(m,2H),7.40(dd,J1=8.5Hz,J2=4.0Hz,1H),7.23-7.13(m,2H) ,7.09(d,J=9.0Hz,1H),6.55(t,J=74Hz,1H),5.79(d,J=8.0Hz,1H),4.73(d,J=3.0Hz,1H) ,4.15(s,1H),3.84-3.72(m,2H),3.02-2.89(m,1H),2.35-2.18(m,1H),1.69-1.57(m,1H).

[0151] Preparation of (3aR,4R,11cR)-4-(4-(2,2,2-trifluoroethoxy)phenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-9)

[0152] Compound V-9 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by 4-(2,2,2-trifluoroethoxy)benzaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 45%; ESI-MS: 401.1 [M+H] + ; 1 H NMR(500MHz,Chloroform-d)δ8.69(dd,J1=4.0Hz,J2=1.5Hz,1H),8.56-8.43(m,1H),7. 88(d,J=9.0Hz,1H),7.53-7.43(m,2H),7.38(dd,J1=8.5Hz,J2=4.0Hz,1H),7.06(d,J=9 .0Hz,1H),7.04-6.93(m,2H),5.79(d,J=8.0Hz,1H),4.70(d,J=3.0Hz,1H),4.39(q,J=8 .0Hz,2H),3.85-3.69(m,2H),3.01-2.89(m,1H),2.37-2.20(m,1H),1.69-1.54(m,1H).

[0153] Preparation of (3aR,4R,11cR)-4-(4-(2,2-difluoroethoxy)phenyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-10)

[0154] Compound V-10 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by 4-(2,2-difluoroethoxy)benzaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 45%; ESI-MS: 383.2 [M+H] + ; 1 H NMR(500MHz,Chloroform-d)δ8.69(dd,J1=4.0Hz,J2=1.5Hz,1H),8.54-8.38(m,1H),7.87(d,J =9.0Hz,1H),7.49-7.40(m,2H),7.37(dd,J1=8.5Hz,J2=4.0Hz,1H),7.06(d,J=9.0Hz,1H),7.0 0-6.94(m,2H),6.28-5.98(m,1H),5.79(d,J=8.0Hz,1H),4.69(d,J=3.0Hz,1H),4.29-4.16(m, 2H),4.09(s,1H),3.85-3.67(m,2H),3.03-2.88(m,1H),2.35-2.20(m,1H),1.67-1.57(m,1H).

[0155] Preparation of (3aR,4R,11cR)-4-(5-chlorofuran-2-yl)-2,3a,4,5,11c-hexahydrofluoro[2,3-a][4,7]phenanthroline (compound V-11)

[0156] Compound V-11 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by 5-chloro-2-furfural and 2-naphthylamine replaced by 6-aminoquinoline, yielding a white solid. Yield 46%; ESI-MS: 327.1 [M+H] + ; 1H NMR(500MHz,Chloroform-d)δ8.69(dd,J1=4.5Hz,J2=1.5Hz,1H),8.52(d,J=8.5Hz,1H),7.93(d,J= 9.0Hz,1H),7.40(dd,J1=8.5Hz,J2=4.5Hz,1H),7.08(d,J=9.0Hz,1H),6.34(dd,J1=3.5Hz,J2=1.5Hz ,1H),6.16(d,J=3.5Hz,1H),5.70(d,J=8.0Hz,1H),4.69(d,J=3.5Hz,1H),4.25(s,1H),3.84(td,J1 =8.0Hz, J2=7.5Hz,1H),3.76-3.65(m,1H),3.20-3.08(m,1H),2.38-2.22(m,1H),2.06-1.92(m,1H).

[0157] Preparation of (3aR,4S,11cR)-4-cyclohexyl-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-12)

[0158] Compound V-12 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by cyclohexaneformaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 56%; ESI-MS: 309.4 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.51 (dd, J1=4.0Hz, J2=1.5Hz, 1H), 8.31-8.26 (m, 1H), 7.62 (d, J=9.0 Hz,1H),7.36-7.29(m,2H),5.73-5.69(m,1H),5.55(d,J=8.0Hz,1H),3.69(q,J=8.0Hz,1H),3 .55-3.49(m,1H),3.07(dd,J1=8.5Hz,J2=2.5Hz,1H),2.85-2.76(m,1H),2.25-2.15(m,1H),1 .92-1.83(m,4H),1.80-1.74(m,2H),1.70-1.62(m,2H),1.32-1.20(m,4H),1.08-0.94(m,2H).

[0159] Preparation of (3aR,4S,11cR)-4-cyclopentyl-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-13)

[0160] Compound V-13 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by cyclopentaneformaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 40%; ESI-MS: 295.3 [M+H] + ; 1 H NMR(500MHz,DMSO-d6)δ8.51(dd,J1=4.0Hz,J2=1.5Hz,1H),8.33-8.24(m,1H),7.62( d,J=9.0Hz,1H),7.36-7.29(m,2H),5.67(d,J=1.5Hz,1H),5.55(d,J=8.0Hz,1H),3.69 (q,J=8.0Hz,1H),3.58-3.53(m,1H),3.10(dd,J1=9.5Hz,J2=2.5Hz,1H),2.77(q,J=9. 5Hz,1H),2.17-2.05(m,1H),1.96-1.79(m,4H),1.71-1.52(m,4H),1.35-1.20(m,3H).

[0161] Preparation of (3aR,4S,11cR)-4-(tetrahydro-2H-pyran-4-yl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-14)

[0162] Compound V-14 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by tetrahydropyran-4-carboxaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 46%; ESI-MS: 311.3 [M+H] + ; 1H NMR(500MHz,DMSO-d6)δ8.52(dd,J1=4.0Hz,J2=1.5Hz,1H),8.30(dd,J1=8.5Hz,J2=1.5Hz,1H),7.63(d,J=9.0Hz,1 H),7.34(dd,J1=8.5Hz,J2=4.0Hz,1H),7.31(d,J=9.0Hz,1H),5.76(s,1H),5.56(d,J=8.0Hz,1H),3.98-3.89(m,2H) ,3.69(q,J=9.0Hz,1H),3.58-3.49(m,1H),3.39-3.35(m,1H),3.33-3.30(m,1H),3.10(dd,J1=9.0Hz,J2=2.5Hz,1H) ,2.85-2.75(m,1H),2.17-2.07(m,1H),1.93-1.84(m,2H),1.77-1.69(m,1H),1.70-1.60(m,1H),1.37-1.21(m,3H).

[0163] Preparation of (3aR,4S,11cR)-4-(piperidin-4-yl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-15)

[0164] Compound V-15 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by piperidine-4-carboxaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 43%; ESI-MS: 310.1 [M+H] + ; 1H NMR(500MHz,DMSO-d6)δ8.53(dd,J1=4.0Hz,J2=1.5Hz,1H),8.31(dd,J1=8.0Hz,J2=1.5Hz,1H),7.65(d,J=9.0Hz,1 H),7.36(dd,J1=8.5Hz,J2=4.0Hz,1H),7.32(d,J=9.0Hz,1H),5.88(s,1H),5.57(d,J=8.0Hz,1H),3.75-3.66(m,1H) ,3.59-3.51(m,1H),3.42-3.35(m,2H),3.13(dd,J1=9.0Hz,J2=2.5Hz,1H),3.10-2.98(m,4H),2.95-2.86(m,2H),2. 84-2.75(m,1H),2.36(d,J=13.5Hz,1H),2.03-1.94(m,1H),1.95-1.80(m,2H),1.79-1.67(m,1H),1.5-1.40(m,2H).

[0165] Preparation of (3aR,4S,11cR)-4-butyl-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-16)

[0166] Compound V-16 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by n-pentanal and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 46%; ESI-MS: 283.3 [M+H] + ; 1 H NMR (500MHz, DMSO-d6) δ8.51 (dd, J1=4.0Hz, J2=1.5Hz, 1H), 8.29-8.26 (m, 1H), 7.63 (d, J=9.0Hz,1H),7.33(dd,J1=8.5Hz,J2=4.0Hz,1H),7.19(d,J=9.0Hz,1H),5.94(s,1H),5 .51(d,J=8.0Hz,1H),3.69(q,J=8.0Hz,1H),3.62-3.54(m,1H),2.73(q,J1=9.0Hz,J2=8 .5Hz,1H),1.93-1.81(m,2H),1.67-1.54(m,2H),1.54-1.42(m,3H),1.41-1.31(m,4H).

[0167] Preparation of (3aR,4S,11cR)-4-pentyl-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-17)

[0168] Compound V-17 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by n-hexanal and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 42%; ESI-MS: 297.4 [M+H] + ; 1 H NMR(500MHz,Chloroform-d)δ8.64(dd,J1=4.0Hz,J2=1.5Hz,1H),8.47-8.44(m,1H),7.82(d,J =9.0Hz,1H),7.34(dd,J1=8.5Hz,J2=4.0Hz,1H),6.99(d,J=9.0Hz,1H),5.62(d,J=8.0Hz,1H),3 .87-3.79(m,1H),3.82-3.74(m,1H),3.49-3.42(m,1H),2.87-2.75(m,1H),2.16-2.03(m,1H), 2.00-1.89(m,1H),1.67-1.54(m,2H),1.51-1.41(m,2H),1.43-1.31(m,5H),0.95-0.90(m,3H).

[0169] Preparation of (3aR,4S,11cR)-4-hexyl-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-18)

[0170] Compound V-18 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by n-heptanal and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 42%; ESI-MS: 311.2 [M+H] + .

[0171] Preparation of (3aR,4S,11cR)-4-heptayl-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-19)

[0172] Compound V-19 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by n-octaldehyde and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 57%; ESI-MS: 325.4 [M+H] + ; 1H NMR(500MHz,Chloroform-d)δ8.63(dd,J1=4.5Hz,J2=1.5Hz,1H),8.60-8.54(m,1H),8 .03-7.92(m,1H),7.42(d,J=6.0Hz,1H),7.04(d,J=9.0Hz,1H),5.61(d,J=8.0Hz,1H), 3.88-3.81(m,1H),3.82-3.74(m,1H),3.52-3.44(m,1H),2.88-2.76(m,1H),2.13-2.0 3(m,1H),2.00-1.92(m,1H),1.42-1.18(m,9H),0.94-0.87(m,3H),0.86-0.77(m,3H).

[0173] Preparation of (3aR,4S,11cR)-4-(perfluorobutyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-20)

[0174] Compound V-20 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by perfluoropentanal and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield: 50%; ESI-MS: 445.3 [M+H] + ; 1 H NMR (500MHz, Chloroform-d)δ

[0175] 8.64(dd,J1=4.0Hz,J2=1.5Hz,1H),8.47-8.41(m,1H),7.81(d,J=9.0Hz,1H),7.34(dd,J1=8.5Hz,J1=4.0Hz,1H),6.98(dd,J1=9.0Hz, J2=2.5Hz,1H),5.62(d,J=8.0Hz,1H),3.95-3.85(m,2H),3.52-3.42(m,2H),2.87-2.76(m,1H),2.16-2.05(m,1H),1.94-1.83(m,1H).

[0176] Preparation of (3aR,4S,11cR)-4-(perfluoropentyl)-2,3,3a,4,5,11c-hexahydrofuran[2,3-a][4,7]phenanthroline (compound V-21)

[0177] Compound V-21 was prepared following the synthesis of I-1, with 4-chlorobenzaldehyde replaced by perfluorohexanal and 2-naphthylamine replaced by 6-aminoquinoline, yielding a yellow solid. Yield 42%; ESI-MS: 495.1 [M+H] +; 1 H NMR (500MHz, Chloroform-d)δ

[0178] 8.68-8.59(m,1H),8.54(d,J=8.5Hz,1H),7.94(d,J=9.0Hz,1H),7.40(dd,J1=8.5Hz,J2=4.5Hz,1H),7.02(dd,J1=9.0Hz,J2=2. 0Hz,1H),5.61(d,J=8.0Hz,1H),5.18-5.10(m,1H),4.37-4.19(m,1H),3.95-3.84(m,2H),2.90-2.74(m,1H),0.85-0.80(m,1H).

[0179] Biological Experiment Example 1: Evaluation Experiment of the Androgen Receptor Antagonistic Ability of the Compounds in the Examples

[0180] This section uses enzalutamide (Enz) as a positive control and employs the LNCaP-ARR2PB-eGFP stable expression model to evaluate the inhibitory activity of some of the compounds in the above examples on the eGFP protein expression of the androgen-dependent prostate cancer cell line LNCaP. Other compounds of the present invention have similar beneficial effects to the compounds listed below, but this should not be construed as the compounds of the present invention having only the following beneficial effects.

[0181] Detection Principle: Androgen receptors (ARRs), as transcription factors, require binding to androgen response elements to exert their transcriptional activity. Therefore, an enhanced green fluorescent protein (eGFP) reporter gene controlled by the ARR2PB promoter was introduced into the androgen-dependent prostate cancer cell line LNCaP to construct a stable LNCaP-ARR2PB-eGFP expression model. After treatment with different concentration gradients of test compounds, the fluorescence intensity value was measured to directly reflect the transcriptional activity of AR, and the IC50 of AR transcriptional inhibition was calculated. 50 The value indicates the strength of the compound's antagonistic ability against androgen receptors; enzalutamide serves as a positive control.

[0182] Detection Procedure: After digesting and centrifuging cells in the logarithmic growth phase, discard the supernatant. Resuspend the cells in 10 mL of phenol red-free RPMI 1640 medium. Mix 20 μL of the cell suspension with an equal volume of 0.2% trypan blue solution. Carefully add 20 μL of this mixture to the wells of a cell counting plate and count the cells. Adjust the cell suspension concentration and add it to the center 60 wells of a 96% empty plate, ensuring each well contains 3.5 x 10⁻⁶ cells. 4Cells were cultured in 90 μL suspensions in each well. The culture plates were placed in a 37°C incubator. The next day, after observing complete cell adhesion, drug administration was initiated. For single-point screening, columns 2-9 were administered eight concentration gradients of the same compound plus 5 nM DHT; column 10 was administered 5 nM DHT as a negative control; and column 11 was administered blank culture medium as a blank control. Three replicates were set up for each compound. After drug administration, the plates were incubated at 37°C for 72 hours. The fluorescence values ​​at 530 nm were detected using a Synergy H1 multi-functional microplate reader under 485 nm excitation light. The average value of the 5 nM DHT wells was taken as 0%, and the blank culture medium group was taken as 100% for normalization of the transcriptional repressive activity of the compound. IC50 was calculated. 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0183] Detection results: As shown in Table 2, most compounds exhibited strong AR transcriptional repression activity and have good application prospects.

[0184] Table 2. AR transcriptional repressive activity of relevant compounds

[0185]

[0186]

[0187] Biological Experiment Example 2: Toxicity of Representative Compounds to Mouse Embryonic Fibroblasts

[0188] Detection Principle: To verify that the aforementioned antagonistic activity does not originate from cytotoxicity but rather specifically targets androgen receptor-dependent prostate cancer cells, this invention employs the aforementioned MTT assay to detect whether the compound inhibits the proliferation of mouse embryonic fibroblasts (NIH-3T3) to assess toxicity. The basic principle of this experiment is that succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to water-insoluble blue-purple formazan crystals, which are then deposited in the cells. Dead cells lack this function. Buffer solution is added to dissolve the formazan formed in the cells, and its absorbance is measured at 490 nm using an enzyme-linked immunosorbent assay (ELISA) reader, which quantitatively reflects the number of living cells. IC50 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0189] Detection steps: Use complete culture medium at 5*10 3NIH-3T3 cells were seeded at a density of cells / well in 96-well plates. After the cells were stably adhered, they were incubated at 37°C for 24 h. Then, the cells were treated with different concentrations of compounds and incubated for another 72 h. Subsequently, 10 μL of 5 mg / mL MTT was added to each well, and the cells were incubated for another 4 h. Then, 100 μL of SDS-HCl-PBS triple buffer was added to each well, and the cells were incubated overnight at 37°C. The absorbance of each well at 570 nm was measured using a microplate reader and converted to the viability.

[0190] Detection results: The results are shown in Table 3. Representative compounds showed low toxicity to normal cells, DC 50 The values ​​are all greater than 50 μM.

[0191] Table 3. Cytotoxicity of representative compounds

[0192] Compound numbering <![CDATA[DC 50 (μM)]]> Compound numbering <![CDATA[DC 50 (μM)]]> Compound numbering <![CDATA[DC 50 (μM)]]> Ⅰ-1 >50 Ⅰ-2 >50 Ⅱ-1 >50 Ⅱ-9 >50 Ⅲ-1 >50 Ⅲ-2 >50 Ⅲ-3 >50 Ⅴ-1 >50 Ⅴ-3 >50 Ⅴ-5 >50 Ⅴ-6 >50 Ⅴ-7 >50 Ⅴ-8 >50 Ⅴ-9 >50 Ⅴ-10 >50 Ⅴ-17 >50 Enz >50

[0193] Biological Experiment Example 3: Inhibitory activity of compounds I-1, II-1, III-3, V-1, V-5, V-6, and V-17 against the proliferation of various prostate cancer cells.

[0194] Detection Principle: This invention also detected the antiproliferative activity of representative compounds in prostate cancer cells. The experiment used was the 3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide (MTT) assay to detect the inhibitory activity of representative compounds I-1, II-1, III-3, V-1, V-5, V-6, and V-17 on the proliferation of androgen-dependent LNCaP cells, as well as androgen receptor-independent PC3 and DU145 cells.

[0195] Detection steps: Use androgen-free complete culture medium at 5*10 3 1.5*10 3 1.5*10 3 and 1.5*10 3 LNCaP, PC3, C4-2, and DU145 cells were seeded at a density of cells / well in 96-well plates. After the cells were stably adhered, 1 nM DHT and different concentrations of test compounds were administered simultaneously. After incubation for 72 h, 10 μL of 5 mg / mL MTT was added to each well, and the cells were incubated for another 4 h. Then, 100 μL of SDS-HCl-PBS triple buffer was added to each well, and the cells were incubated overnight at 37°C. The absorbance at 570 nm was measured using a microplate reader and converted to the viability. IC50 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0196] Test results: The results are as follows Figure 1Compounds I-1, II-1, III-3, V-1, V-5, V-6, and V-17 showed concentration-dependent inhibitory effects on the androgen-dependent cell line LNCaP, comparable to those of enzalutamide; Figure 2 , Figure 3 As shown, similar to enzalutamide, compounds I-1, II-1, III-3, V-1, V-5, V-6 and V-17 did not significantly inhibit the proliferation of androgen-independent cell lines DU145 and PC3 at concentrations of 16.7 μM and below, but showed some killing effect at a concentration of 50 μM, demonstrating that they selectively act on AR-related pathways.

[0197] Biological Experiment Example 4: Dual-luciferase reporter gene assay for the effects of compounds I-1, II-1, III-3, and V-1 on ARLBD F877L / T878A Double mutation activity

[0198] Detection Principle: This is a reporter system that uses luciferin as a substrate to detect the activity of luciferase in fireflies. It quantifies luciferase activity by detecting fluorescence intensity, and further quantifies the interaction between transcription factors and target promoters. AR LBD is employed. F877L / T878A A double-mutant enzalutamide resistance model was used to evaluate the transcriptional repression activity of compounds I-1, II-1, III-3, and V-1, with dalolutamide (abbreviated as Daro) as a positive control.

[0199] Detection method: After digesting and centrifuging cells in the logarithmic growth phase, resuspend the cells in 10 mL of phenol red-free RPMI 1640 medium and count them. Adjust the cell suspension concentration and seed them into 96-well plates with a white background, at a volume of 75 μL per 10,000 cells. Incubate at 37°C. After 24 h, transfect according to the transfection reagent instructions. Add 0.1 μg of DNA and 0.25 μL of transfection reagent to each well, pre-mixed with 25 μL of Opti-MEM serum-depleted medium and incubated for 30 min before adding to the 96-well plate. The co-transfected DNA was divided into three categories: the first category was plasmids expressing nuclear receptors; the second category was plasmids containing the corresponding promoters expressing firefly luciferase; and the third category was Renilla plasmids expressing Renilla luciferase. The Renilla transfection ratio was 1 / 20 of the total DNA, and the ratio of nuclear receptor to its corresponding promoter plasmid was 3:1. The samples were placed in an incubator for static incubation (pGL4.18-ARR2PB-Luc uses the ARF876L / T877A promoter, and pGL4.18-mmTV-Luc uses the MR and PRDRAM promoters). After 24 hours, the culture medium containing the transfection reagent was aspirated, and 90 μL of fresh phenol red-free 1640 medium was added to each well. A concentration gradient of the drug and corresponding hormone was added to each well, with a hormone control group and a blank control group included. 24 hours after drug administration, fluorescence values ​​were measured according to the kit instructions. The ratio of firefly luciferase to Renilla luciferase in each well was calculated, and the activator / inhibitor ratio of the corresponding compound was calculated. IC50 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0200] Test results: The results are as follows Figure 4 As shown, compounds I-1, II-1, III-3, and V-1 are effective in the drug resistance model and can inhibit AR in a concentration-dependent manner. F877L / T878A The mutant's transcriptional activity was comparable to that of dalolutamide. Furthermore, enzalutamide acts as an AR... F877L / T878A Due to the partial agonist activity of the mutant, the IC50 of enzalutamide in antagonistic mode cannot be inferred. 50 value.

[0201] Biological Experiment Example 5: Represents the inhibitory activity of compounds II-1 and V-1 on the androgen receptor target gene PSA protein.

[0202] Detection principle: Prostate-specific antigen (PSA) is one of the key indicators for clinical detection of prostate cancer. Its expression is positively correlated with the transcription level of androgen receptor. In order to further verify the relevant mechanism of the antagonistic effect of compounds II-1 and V-1, the downstream signaling pathway of AR was evaluated and the level of PSA secreted by LNCaP cells was measured.

[0203] Detection method: The detection method for secretory PSA is the same as that for evaluating AR transcriptional repression activity (refer to Biological Experiment Example 1), except that the volume of each well is set to 180 μL during plate preparation. 72 h after drug administration, the supernatant from each well is collected and sent to the Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital) to measure PSA levels and IC50. 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0204] Test results: The results are as follows Figure 5 As shown, the results of the exo-PSA assay showed that compounds II-1 and V-1 could inhibit PSA exo-secretion from LNCaP cells in a concentration-dependent manner, with an inhibitory effect comparable to that of enzalutamide.

[0205] Biological Experiment Example 6: Competitive binding experiment of androgen receptors representing compounds II-1 and V-1 to verify the binding target.

[0206] Detection principle: Utilizing Polar Screen TM The Androgen Receptor Competitor Assay Kit tests compounds using a green fluorescent ligand that binds strongly to the AR LBP site. When the test compound, the ligand, and the protein are added to the same system, the compound and ligand competitively bind to the AR LBP site. If the compound enters the LBP pocket, the ligand tracer is released from the complex, resulting in a decrease in polarization. Therefore, the change in fluorescence polarization in this system determines the relative affinity of the test compound for the AR LBP.

[0207] Detection Procedure: Androgen receptor ligand domain protein and high-affinity fluorescent ligand were mixed in buffer, and different concentrations of test compounds II-1 and V-1 were added, with dihydrotestosterone (DHT) used as a positive control. Changes in fluorescence polarization values ​​were measured using a multi-mode microplate reader to quantitatively determine whether the antagonistic compounds targeted the androgen receptor. IC50 50 The values ​​were obtained by fitting using Graphpad Prism 8.0.2 software.

[0208] Test results: The results are as follows Figure 6 As shown, compounds II-1 and V-1 can correctly target the androgen receptor ligand domain at different concentrations and exhibit different binding strengths, with their binding affinity being comparable to that of the positive enzalutamide.

[0209] Biological Experiment Example 7: X-ray diffraction pattern of compound III-2 (single crystal)

[0210] Detection principle: When a crystal is irradiated with X-rays, the scattered X-rays from each atom in the crystal will superimpose. When the X-rays are monochromatic, the scattered X-rays from each atom interfere with each other, producing strong X-ray diffraction lines in a specific direction. The structure of compound III-2 crystal is determined by utilizing the X-ray diffraction effect of a single crystal.

[0211] Detection results: The X-ray diffraction pattern of compound Ⅲ-2 is as follows: Figure 7 As shown, its absolute configuration is (3aR,4aR,11cR)-4-(4-bromophenyl)-2,3,3a,4,5,11c-hexahydrophenylpropane[f]furan[3,2c]quinoline, which is consistent with the theory.

Claims

1. A substituted tetrahydroquinoline derivative, characterized in that, Selected from the following compounds: 。 2. The use of the substituted tetrahydroquinoline derivative of claim 1 in the preparation of androgen receptor antagonists, characterized in that, Application in the preparation of drugs for treating diseases related to abnormal expression or mutation of androgen receptors.

3. The application according to claim 2, characterized in that, The disease mentioned is prostate cancer.

4. The application according to claim 3, characterized in that, The prostate cancer referred to includes metastatic prostate cancer, castration-resistant prostate cancer, and metastatic castration-resistant prostate cancer.

5. The application according to claim 2, characterized in that, The disease mentioned is breast cancer.

6. The application according to claim 2, characterized in that, The disease mentioned is ovarian cancer.

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