Heterocyclic compound, pharmaceutical composition, and use of heterocyclic compound or pharmaceutical composition

Abstract

Disclosed in the present invention are a heterocyclic compound, a pharmaceutical composition, and a use of the heterocyclic compound or the pharmaceutical composition. Provided is a heterocyclic compound, or a pharmaceutically acceptable salt, stereoisomer, tautomer, polymorphic substance, solvate, isotope derivative, or prodrug thereof. The heterocyclic compound has a structure represented by formula (I). The heterocyclic compound provided in the present invention has inhibitory activity against a TEAD protein, can effectively inhibit the interaction between TEAD and YAP, and thus can treat diseases related to abnormal regulation of the TEAD protein or an abnormal Hippo pathway.

Description

A heterocyclic compound, a pharmaceutical composition and its application Technical Field

[0001] This invention relates to the field of small molecule compound technology, and in particular to a heterocyclic compound, a pharmaceutical composition and its application. Background Technology

[0002] The Hippo signaling pathway is highly conserved in higher vertebrates and participates in regulating many biological processes, such as cell proliferation, survival, differentiation, and organ size regulation. Studies have shown that abnormal activity of the Hippo signaling pathway is often highly correlated with the occurrence of malignant tumors, such as lung cancer, liver cancer, and pancreatic cancer. Therefore, regulating the Hippo signaling pathway has great potential for cancer treatment. YAP / TAZ is the most important downstream effector of the Hippo signaling pathway, capable of regulating the expression of numerous Hippo-related genes. However, due to its disordered and open structural characteristics and lack of active catalytic sites, YAP / TAZ is not an ideal drug target. YAP / TAZ itself cannot interact directly with DNA; it needs to bind to the transcription factor TEAD to initiate downstream gene transcription. Therefore, blocking TEAD-YAP / TAZ binding is widely considered a more promising way to regulate Hippo transcriptional activity. Thus, inhibiting the interactions between YAP, TAZ, TEAD, and YAP-TEAD or TAZ-TEAD proteins through drug intervention appears to be a strategy for the prevention and / or treatment of cancer and other hyperproliferative diseases. Currently, there are no approved drugs for the marketing of TEAD binding agents, YAP-TEAD, or TAZ-TEAD protein-protein interaction inhibitors.

[0003] Therefore, there is still a need to develop safe and effective inhibitors to inhibit the interaction between YAP-TEAD or TAZ-TEAD proteins. Summary of the Invention

[0004] The present invention aims to at least solve one of the technical problems existing in the prior art. To this end, a first aspect of the present invention provides a heterocyclic compound that can be used as an inhibitor to suppress the interaction between TEAD and YAP / TAZ.

[0005] A second aspect of the present invention also provides a pharmaceutical composition.

[0006] A third aspect of the present invention also provides the use of a heterocyclic compound or pharmaceutical composition.

[0007] According to a first aspect of the present invention, a heterocyclic compound, or a pharmaceutically acceptable salt thereof, its stereoisomers, its tautomers, its polymorphs, its solvates, its isotopic derivatives, or its prodrug, has a structure shown in formula (I):

[0008] Where X is N, -CH, or CBr; W is O or S;

[0009] L is Or it may not exist;

[0010] R1 is selected from phenyl, C 2～5 Ester-substituted phenyl, pyrimidinyl, cyclohexyl, naphthyl, and adamantyl groups;

[0011] R2 is selected from H, halogens, and C. 1～5 aldehyde group, carboxyl group, C 1～10 alkyl, C 1～10 Halogenated alkyl, C 1～10 Alkoxy,

[0012] Among them, R5 and R6 are independently selected from H, Alternatively, R5 and R6 can form a saturated six-membered heterocycle doped with nitrogen and sulfur; V is selected from C or N;

[0013] R7 and R8 are independently selected from H. Alternatively, R7 and R8 can form a saturated six-membered heterocycle doped with nitrogen and sulfur;

[0014] R3 is selected from H, halogens, and C. 1～10 alkyl, C 1～10 Halogenated alkyl, C 1～10 Alkoxy,

[0015] R4 is selected from C 1～10 Alkyl, phenyl or C 1～10 Halogenated alkyl, C 1～10 Halogenated alkoxy groups, fluorinated sulfo-substituted phenyl groups, and C 1～10 Halogenated alkyl, C 1～10 Benzyl groups substituted with halogenated alkoxy groups.

[0016] The heterocyclic compounds according to embodiments of the present invention have at least the following beneficial effects:

[0017] The heterocyclic compound provided by this invention exhibits strong binding activity to TEAD protein. After binding to TEAD protein, the compound significantly enhances the protein's thermal stability while inhibiting TEAD protein activity. It effectively disrupts the interaction between TEAD and YAP, and demonstrates strong anti-proliferative activity against the NCI-H226 cell line with NF2 deficiency and Hippo pathway abnormalities. This allows for the treatment of diseases related to abnormal TEAD protein regulation or Hippo pathway abnormalities. Compared with related TEAD-targeting anti-tumor compounds such as MYF-01-037 (Cancer Cell. 2020, 37(1), 104-122. J. Med. Chem. 2022 65(13): 9206-9229.), the compound of this invention shows significantly improved in vitro, cellular, and animal-level anti-tumor activity, making it an effective in vivo TEAD-targeting inhibitor. It holds promise for development into a novel, original anti-tumor drug.

[0018] According to some embodiments of the present invention, R2 is selected from H, aldehyde,

[0019] Among them, R5 and R6 are independently selected from H, Alternatively, R5 and R6 can form a saturated six-membered heterocycle doped with nitrogen and sulfur; V is selected from C or N;

[0020] R7 and R8 are independently selected from H. Alternatively, R7 and R8 can form a saturated six-membered heterocycle doped with nitrogen and sulfur.

[0021] According to some embodiments of the present invention, the nitrogen- and sulfur-doped saturated six-membered heterocycles include 1-thiomorpholine oxide and 1,1-thiomorpholine dioxide.

[0022] According to some embodiments of the present invention, R3 is selected from H, halogens,

[0023] According to some embodiments of the present invention, the heterocyclic compound is selected from one of the following structural formulas:

[0024] According to some embodiments of the present invention, the small molecule compounds of the present invention have an asymmetric center, a chiral axis, and a chiral plane, and can exist in the form of a racemic mixture, an R-isomer, or an S-isomer. Those skilled in the art can obtain the R-isomer and / or S-isomer from the racemic mixture using conventional techniques.

[0025] According to a second aspect of the present invention, a method for preparing a heterocyclic compound is provided, comprising the following steps:

[0026] S1. Compound B is mixed with an acidic reagent and deprotected to obtain an intermediate;

[0027] S2. The intermediate, compound A and (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) are mixed and reacted to obtain the product.

[0028] Alternatively, compound A and compound C react to obtain the product.

[0029] Alternatively, compound D and compound E can be reacted to obtain the product.

[0030] The structural formulas of compounds A, B, C, D, and E are as follows:

[0031] According to some embodiments of the present invention, the acidic reagent includes trifluoroacetic acid and / or hydrochloric acid.

[0032] According to some embodiments of the present invention, in step S1, the molar ratio of compound B to acidic reagent is 1:(0.5-2).

[0033] According to some embodiments of the present invention, in step S2, the molar ratio of compound A and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate is 1:(0.8-1.5).

[0034] According to some embodiments of the present invention, in step S2, the temperature of the reaction is 20–35°C.

[0035] According to some embodiments of the present invention, in step S2, the reaction time is 3 to 8 hours.

[0036] According to a third aspect of the present invention, a pharmaceutical composition is provided comprising the heterocyclic compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, its stereoisomer, its tautomer, its polymorph, its solvate, its isotopic derivative, or its prodrug; and a pharmaceutically acceptable excipient.

[0037] According to some embodiments of the present invention, the excipients include at least one of solvents, excipients, diluents, binders, disintegrants, dispersants, flavoring agents, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, preservatives, solid binders, flow aids, or lubricants.

[0038] The third aspect of the present invention provides a heterocyclic compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, its stereoisomer, its tautomer, its polymorph, its solvate, its isotopic derivative, or its prodrug; or the use of the pharmaceutical composition as described in the third aspect of the present invention in the preparation of a medicament for the prevention and / or treatment of TEAD-mediated diseases or conditions.

[0039] According to some embodiments of the present invention, the diseases or conditions mediated by TEAD are selected from colon cancer, diffuse large B-cell lymphoma, follicular lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate cancer, breast cancer, bile duct and gallbladder cancer, bladder cancer; brain tumors, neuroblastoma, schwannoma, glioma, glioblastoma and astrocytoma; cervical cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal carcinoma, ovarian cancer, pancreatic cancer, renal cell carcinoma, rectal cancer, thyroid cancer, parathyroid tumors, uterine tumors and soft tissue sarcomas, cardiovascular diseases, neurodegenerative diseases, malaria, AIDS, gout, diabetes, renal failure, and chronic lung diseases.

[0040] Definitions and general terms

[0041] "Halogen" includes any one or more of fluorine, chlorine, bromine, and iodine.

[0042] “C 1～10 "alkyl" refers to an alkyl group with a total number of carbon atoms of 1 to 10, including C14 and C24. 1-10 straight-chain alkyl, C 1-10 Branched alkyl groups and C 3-10 The cycloalkyl group can be, for example, a straight-chain alkyl group with a total number of carbon atoms of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; a branched-chain alkyl group with a total number of carbon atoms of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; or a cycloalkyl group with a total number of carbon atoms of 3, 4, 5, 6, 7, 8, 9, or 10, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, cyclopropyl, methylcyclopropyl, ethylcyclopropyl, cyclopentyl, methylcyclopentyl, cyclohexyl, etc. wait.

[0043] “C 1～10 "halogenated alkyl" and C 1～10 Alkyl groups have similar definitions, except that optionally at least one H in the group is substituted with a halogen, such as -CF3.

[0044] “C 1～10"Alkoxy" refers to alkoxy groups with a total number of carbon atoms of 1-10, including C1-10 straight-chain alkoxy groups, C1-10 branched-chain alkoxy groups, and C2-10 cycloalkoxy groups. For example, it can be a straight-chain alkoxy group with a total number of carbon atoms of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; a branched-chain alkoxy group with a total number of carbon atoms of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; or a cycloalkoxy group with a total number of carbon atoms of 2, 3, 4, 5, 6, 7, 8, 9, or 10. For example, it can be methoxy, ethoxy, n-propoxy, isopropoxy, etc.

[0045] “C 1～10 "haloalkoxy" and "C" 1～10 The alkoxy group has a similar definition, except that at least one H in the group is optionally replaced by a halogen. For example, -OCF3.

[0046] When L in this invention is selected At that time, the carbonyl group in L is attached to the phenyl group.

[0047] The pharmaceutically acceptable salts described in this invention can be salts formed by anion and a positively charged group on a compound of Formula I. Suitable anions include chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronate, lactate, glutarate, or maleate. Similarly, salts can be formed by cations and negatively charged groups on a compound of Formula I. Suitable cations include sodium, potassium, magnesium, calcium, and ammonium ions, such as tetramethylammonium ions.

[0048] "Pharmaceutically acceptable salts" refer to salts formed by compounds of Formula I with acids selected from the group consisting of: hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pyric acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, and hydroxyethanesulfonic acid; or sodium, potassium, calcium, aluminum, or ammonium salts formed by compounds of Formula I with inorganic bases; or methylamine, ethylamine, or ethanolamine salts formed by compounds of general Formula I with organic bases.

[0049] There are no particular limitations on the administration of the active ingredients or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), etc.

[0050] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.

[0051] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures thereof. Besides these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances.

[0052] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0053] The compounds of this invention can be administered alone or in combination with other therapeutic agents (such as antitumor drugs).

[0054] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 20–500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skills of a skilled physician.

[0055] Other features and advantages of the invention will be set forth in the description which follows, and will be apparent in part from the description, or may be learned by practicing the invention. Attached Figure Description

[0056] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0057] Figure 1a shows the IC50 values ​​of the compound prepared in Example 1 against the inhibitory activity of TEAD1 protein from the four TEAD families. 50 value;

[0058] Figure 1b shows the IC50 values ​​of the compound prepared in Example 1 against the inhibitory activity of TEAD2 protein from the four TEAD families. 50 value;

[0059] Figure 1c shows the IC50 values ​​of the compound prepared in Example 1 against the inhibitory activity of TEAD4 protein from the four TEAD families. 50 value. Detailed Implementation

[0060] The following are specific embodiments of the present invention, and the technical solutions of the present invention will be further described in conjunction with the embodiments, but the present invention is not limited to these embodiments.

[0061] Unless otherwise specified, the reagents, methods and equipment used in this invention are all conventional reagents, methods and equipment in this technical field.

[0062] The raw materials used in this invention are as follows:

[0063] The reaction equation and preparation method of compound B-1 are as follows:

[0064] Step 1: Aniline B-1a (9.3 g, 100.0 mmol, 1.0 equivalent) was dissolved in 100 mL of pyridine, and then phenyl chloroformate (14.0 mL, 100.0 mmol, 1.0 equivalent) was slowly added dropwise. The mixture was stirred overnight at room temperature. After thin-layer chromatography showed that the reaction was complete, 100 mL of 10% citric acid aqueous solution was added to quench the reaction. The mixture was then extracted three times with 180 mL of ethyl acetate. The combined organic phases were washed with saturated sodium bicarbonate aqueous solution, saturated sodium chloride aqueous solution, and water. After drying with anhydrous sodium sulfate, the mixture was concentrated under vacuum to obtain 17.1 g of crude product compound B-1b, which could be used directly in the next reaction without purification.

[0065] Step 2: Aniline B-1b (17.1 g, 80.0 mmol, 1.0 equivalent) was dissolved in 100 mL of dimethyl sulfoxide, followed by the addition of tert-butylpiperazine carboxylate (15.0 g, 80.0 mmol, 1.0 equivalent) and sodium hydroxide aqueous solution (8.0 mL, 80.0 mmol, 10 N, 1.0 equivalent). The mixture was stirred at room temperature for 2 hours. After the reaction was monitored by HPLC-MS until complete, the reaction was quenched with 100 mL of aqueous solution, followed by extraction three times with 180 mL of ethyl acetate. The combined organic phases were washed with saturated sodium bicarbonate aqueous solution, saturated sodium chloride aqueous solution, and water. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The purified compound was then separated and purified by a Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 30%) to obtain 19.5 g of white solid compound B-1, with a yield of 80.0%.

[0066] The NMR data of the product are as follows:

[0067] 1H NMR (500MHz, CDCl3) δ7.36 (d, J=7.4Hz, 2H), 7.32–7.28 (m, 2H), 7.06 (t, J=7.4Hz, 1H), 6.51 (s, 1H), 3.49 (s, 8H), 1.50 (s, 9H). LC-MS (ESI): m / z 306.3[M+H] + .

[0068] Example 1

[0069] This example provides a heterocyclic compound I-1, whose reaction equation and preparation method are as follows:

[0070] Step 1: Dissolve indole-4-carboxylic acid methyl ester (1.75 g, 10.0 mmol, 1.0 equivalent), cuprous iodide (1.9 g, 10.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (882.0 mg, 10.0 mmol, 1.0 equivalent), potassium carbonate (2.76 g, 20.0 mmol, 2.0 equivalent), and 4-iodotrifluorotoluene (2.72 g, 10.0 mmol, 1.0 equivalent) in 25 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was complete as monitored by HPLC-MS, 50 mL of aqueous solution was added to quench the reaction, and then the mixture was extracted three times with 120 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 50%) to give 2.2 g of white solid compound 1b, with a yield of 68.5%. 1 HNMR (500MHz, CDCl3) δ8.01(d,J=8.3Hz,1H),7.84(d,J=8.3Hz,2H),7.78(d,J=8.2Hz,1H),7.65(d,J=8.2 Hz,2H),7.50(d,J=3.3Hz,1H),7.42(d,J=3.3Hz,1H),7.33(t,J=7.9Hz,1H),4.04(s,3H).LC-MS(ESI):m / z 320.3[M+H] + .

[0071] Step 2: Dissolve 1a (2.2 g, 6.9 mmol, 1.0 equivalent) in 30 mL of ethanol, then add potassium hydroxide aqueous solution (0.69 mL, 6.9 mmol, 10 N, 1.0 equivalent). Stir overnight at room temperature. After the reaction is complete as monitored by HPLC-MS, the solvent is removed by vacuum concentration. The pH of the solution is adjusted to 1 with 6 N hydrochloric acid, and a precipitate forms. The solvent is removed by filtration, and the residue is dried to obtain 2.0 g of pink solid compound 1c, with a yield of 95.6%. No purification is required; it can be used directly in the next reaction. LC-MS (ESI): m / z 306.2 [M+H] + .

[0072] Step 3: Dissolve compound B-1 (183.8 mg, 0.6 mmol, 1.2 equivalents) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following the addition of N,N-dimethylformamide, 1c (153.1 mg, 0.5 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (190.0 mg, 0.5 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 30%) to obtain 80.0 mg of white solid compound I-1, with a yield of 32.5%.

[0073] The NMR data of the product are as follows:

[0074] 1 H NMR (500MHz, CDCl3) δ7.86–7.81(m,2H),7.68–7.62(m,3H),7.44(d,J=3.3Hz,1H),7.39–7.35(m,2H),7.35–7.29(m,3 H),7.27–7.23(m,1H),7.07(t,J=7.3Hz,1H),6.75(d,J=3.3Hz,1H),6.59(s,1H),4.05–3.42(m,8H).LC-MS(ESI):m / z 493.2[M+H] + .

[0075] Example 2

[0076] This example provides a heterocyclic compound I-2, the reaction equation and preparation method of which are as follows:

[0077] Step 1: Dissolve indole-4-carboxylic acid methyl ester (175.0 mg, 1.0 mmol, 1.0 equivalent), cuprous iodide (190.0 mg, 1.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (88.2 mg, 1.0 mmol, 1.0 equivalent), potassium carbonate (276.0 mg, 2.0 mmol, 2.0 equivalent), and 1-iodo-4-(trifluoromethoxy)benzene (288.0 mg, 1.0 mmol, 1.0 equivalent) in 3 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was complete as monitored by HPLC-MS, 6 mL of aqueous solution was added to quench the reaction, followed by extraction with 18 mL of ethyl acetate three times. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound 2b was obtained by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 50%), yielding 211.0 mg of white solid compound 2b, with a yield of 68.5%. LC-MS (ESI): m / z 336.2 [M+H] + .

[0078] Step 2: Compound 2b (211.0 mg, 0.63 mmol, 1.0 equivalent) was dissolved in 5 mL of ethanol, and then potassium hydroxide aqueous solution (0.063 mL, 0.63 mmol, 10 N, 1.0 equivalent) was added. The mixture was stirred overnight at room temperature. After the reaction was monitored by HPLC-MS to be complete, the solvent was removed by vacuum concentration. The pH of the solution was adjusted to 1 with 6 N hydrochloric acid, and a precipitate was formed. The solvent was removed by filtration, and the filter residue was dried to obtain 188.0 mg of white solid compound 2c, with a yield of 93.2%. No purification was required, and it could be used directly for the next step of the reaction. 1 H NMR (500MHz, CDCl3) δ8.09(d,J=7.5Hz,1H),7.76(d,J=8.2Hz,1H),7.56(d,J=8.4H z,2H),7.49(s,1H),7.43(d,J=9.0Hz,3H),7.34(t,J=7.8Hz,1H).LC-MS(ESI):m / z 322.2[M+H] + .

[0079] Step 3: Dissolve compound B-1 (183.8 mg, 0.6 mmol, 1.01 equivalents) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following the addition of N,N-dimethylformamide, 2c (188.0 mg, 0.59 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (228.0 mg, 0.60 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 30%) to obtain 76.0 mg of white solid compound I-2, with a yield of 24.9%.

[0080] The NMR data of the product are as follows:

[0081] 1 H NMR (500MHz, CDCl3) δ7.61–7.50(m,3H),7.47–7.34(m,5H),7.33–7.28(m,3H),7.22(d,J=7.2Hz,1H ),7.07(t,J=6.9Hz,1H),6.71(s,1H),6.62(s,1H),3.96–3.41(m,8H).LC-MS(ESI):m / z:509.2[M+H] + .

[0082] Example 3

[0083] This example provides a heterocyclic compound I-3, whose reaction equation and preparation method are as follows:

[0084] Step 1: Dissolve methyl indole-4-carboxylate-3-carboxaldehyde (203.2 mg, 1.0 mmol, 1.0 equivalent), cuprous iodide (190.0 mg, 1.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (88.2 mg, 1.0 mmol, 1.0 equivalent), potassium carbonate (276.0 mg, 2.0 mmol, 2.0 equivalent), and 4-iodotrifluorotoluene (272.0 mg, 1.0 mmol, 1.0 equivalent) in 3 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was complete as monitored by HPLC-MS, 6 mL of aqueous solution was added to quench the reaction, and then the mixture was extracted three times with 18 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 50%) to give 184.3 mg of pale yellow solid compound 3b, with a yield of 53.0%. 1 H NMR(500MHz, CDCl3)δ10.52(d,J=6.8Hz,1H),8.15(d,J=4.6Hz,1H),7.93–7.81(m, 3H),7.69–7.60(m,3H),7.39–7.32(m,1H),4.00(d,J=4.6Hz,3H).LC-MS(ESI):m / z 348.1[M+H] + .

[0085] Step 2: Dissolve 3b (184.3 mg, 0.53 mmol, 1.0 equivalent) in 5 mL of ethanol, then add potassium hydroxide aqueous solution (0.053 mL, 0.53 mmol, 10 N, 1.0 equivalent). Stir overnight at room temperature. After the reaction is complete as monitored by HPLC-MS, the solvent is removed by vacuum concentration. The pH of the solution is adjusted to 1 with 6 N hydrochloric acid, and a precipitate forms. The solvent is removed by filtration, and the residue is dried to obtain 173.7 mg of brown solid compound 3c, with a yield of 98.4%. No purification is required; it can be used directly in the next reaction. LC-MS (ESI): m / z 334.1 [M+H] + .

[0086] Step 3: Dissolve compound B-1 (183.8 mg, 0.6 mmol, 1.15 equivalents) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following the addition of N,N-dimethylformamide, 1c (173.7 mg, 0.52 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (228.0 mg, 0.6 mmol, 1.15 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 30%) to obtain 54.0 mg of white solid compound I-3, yield 20.0%.

[0087] The NMR data of the product are as follows:

[0088] 1 H NMR (500MHz, CDCl3) δ10.02(s,1H),8.03(s,1H),7.90(d,J=8.3Hz,2H),7.68(d,J=8.3Hz,2H),7.54(d,J=8.5Hz,1H),7.46–7.41(m,1H ),7.35(d,J=7.2Hz,2H),7.30(d,J=7.2Hz,1H),7.28–7.23(m,2H),7.06–6.97(m,1H),6.84(s,1H),4.18–3.33(m,8H).LC-MS(ESI):m / z 521.2[M+H] + .

[0089] Example 4

[0090] This example provides a heterocyclic compound I-4, whose reaction equation and preparation method are as follows:

[0091] Step 1: Dissolve indole-4-carboxylic acid methyl ester (175.0 mg, 1.0 mmol, 1.0 equivalent), cuprous iodide (190.0 mg, 1.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (88.2 mg, 1.0 mmol, 1.0 equivalent), potassium carbonate (276.0 mg, 2.0 mmol, 2.0 equivalent), and 4-iodobenzenepentafluoride (330.0 mg, 1.0 mmol, 1.0 equivalent) in 3 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was monitored by HPLC-MS to be complete, 6 mL of aqueous solution was added to quench the reaction, and then the mixture was extracted three times with 18 mL of ethyl acetate. The organic phases were combined, washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 50%) to give 276.7 mg of white solid compound 4b, with a yield of 73.2%. 1 H NMR (500MHz, CDCl3) δ8.02(d,J=7.5Hz,1H),7.94(d,J=9.0Hz,2H),7.77(d,J=8.3Hz,1H),7. 58(d,J=8.7Hz,2H),7.47–7.42(m,2H),7.32(t,J=7.9Hz,1H),4.04(s,3H).LC-MS(ESI):m / z 378.1[M+H] + .

[0092] Step 2: Dissolve 4b (276.7 mg, 0.73 mmol, 1.0 equivalent) in 5 mL of ethanol, then add potassium hydroxide aqueous solution (0.073 mL, 0.73 mmol, 10 N, 1.0 equivalent). Stir overnight at room temperature. After the reaction is complete as monitored by HPLC-MS, the solvent is removed by vacuum concentration. The pH of the solution is adjusted to 1 with 6 N hydrochloric acid, and a precipitate forms. The solvent is removed by filtration, and the residue is dried to obtain 239.9 mg of white solid compound 4c, with a yield of 90.5%. No purification is required; it can be used directly in the next reaction. LC-MS (ESI): m / z 364.2 [M+H] + .

[0093] Step 3: Dissolve compound B-1 (198.2 mg, 0.65 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following the addition of N,N-dimethylformamide, 4c (239.9 mg, 0.65 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (247.0 mg, 0.65 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 30%) to give 75.6 mg of white solid compound I-4, with a yield of 21.1%.

[0094] The NMR data of the product are as follows:

[0095] 1 H NMR(500MHz, CDCl3)δ7.96(d,J=8.5Hz,2H),7.70–7.60(m,3H),7.44–7.30(m,6H),7 .08(t,J=7.3Hz,1H),6.76(s,1H),6.60(s,1H),4.00–3.44(m,8H).LC-MS(ESI):m / z 551.0[M+H] + .

[0096] Example 5

[0097] This example provides a heterocyclic compound I-5, whose reaction equation and preparation method are as follows:

[0098] Step 1: Dissolve methyl 6-bromo-4-indolecarboxylate (254.0 mg, 1.0 mmol, 1.0 equivalent), cuprous iodide (190.0 mg, 1.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (88.2 mg, 1.0 mmol, 1.0 equivalent), potassium carbonate (276.0 mg, 2.0 mmol, 2.0 equivalent), and 4-iodotrifluorotoluene (272.0 mg, 1.0 mmol, 1.0 equivalent) in 3 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was complete as monitored by HPLC-MS, 6 mL of aqueous solution was added to quench the reaction, followed by extraction with 18 mL of ethyl acetate three times. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound was then separated and purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 50%) to give 161.2 mg of yellow solid compound 5b, with a yield of 40.5%. LC-MS (ESI): m / z 399.1 [M+H] + .

[0099] Step 2: Dissolve 5b (161.2 mg, 0.45 mmol, 1.0 equivalent) in 5 mL of ethanol, then add potassium hydroxide aqueous solution (0.045 mL, 0.45 mmol, 10 N, 1.0 equivalent). Stir overnight at room temperature. After the reaction is complete as monitored by HPLC-MS, the solvent is removed by vacuum concentration. The pH of the solution is adjusted to 1 with 6 N hydrochloric acid, and a precipitate is formed. The solvent is removed by filtration, and the filter residue is dried to obtain 164.7 mg of white solid compound 5c, with a yield of 95.3%. It can be used directly in the next step of the reaction without purification. 1 H NMR (500MHz, DMSO-d6) δ7.90 (d, J = 8.4Hz, 2H), 7.80 (d, J = 8.4Hz, 2H), 7.50 (d, J = 3.4Hz, 1H), 6.97–6.82 (m, 3H), 6.32 (d, J = 7.5Hz, 1H). LC-MS (ESI): m / z 385.1[M+H] + .

[0100] Step 3: Dissolve compound B-1 (153.3 mg, 0.5 mmol, 1.2 equivalents) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. N,N-Dimethylformamide was followed by the addition of 5c (153.1 mg, 0.43 mmol, 1.0 equivalent), and the pH of the solution was adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (190.0 mg, 0.50 mmol, 1.2 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to be complete, 5 mL of aqueous solution was added to quench the reaction, and the mixture was extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 30%) to give 45.2 mg of white solid compound I-5, with a yield of 18.4%.

[0101] The NMR data of the product are as follows:

[0102] 1 H NMR(500MHz, CDCl3)δ7.85(d,J=8.3Hz,2H),7.61(d,J=8.3Hz,2H),7.52(s,1H),7.37–7.33(m,3 H),7.31–7.28(m,2H),7.06(t,J=7.3Hz,1H),6.81(s,1H),6.71–6.65(m,1H),3.93–3.42(m,8H). LC-MS(ESI):m / z:572.4[M+H] + .

[0103] Example 6

[0104] This example provides a heterocyclic compound I-6, whose reaction equation and preparation method are as follows:

[0105] Step 1: Dissolve intermediate 1c (76.3 mg, 0.25 mmol, 1.0 equivalent) and 5-aminopyrimidine (23.8 mg, 0.25 mmol, 1.0 equivalent) in 2 mL Following N,N-dimethylformamide, N,N-diisopropylethylamine (65.0 mg, 0.50 mmol, 2.0 equivalents) was added and stirred at room temperature for 15 minutes. Then, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (95.0 mg, 0.25 mmol, 102 equivalents) was added and stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS until complete, the reaction was quenched with 3 mL of aqueous solution and extracted three times with 15 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound was then separated and purified by a Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 60%) to give 30.0 mg of white solid compound I-6, with a yield of 31.3%.

[0106] The NMR data of the product are as follows:

[0107] 1 H NMR (500MHz, CDCl3) δ9.33–9.08(m,2H),8.01(s,1H),7.86(d,J=8.3Hz,2H),7.79(d,J=8. 3Hz, 1H), 7.72–7.64 (m, 3H), 7.59–7.55 (m, 1H), 7.39 (t, J = 7.8Hz, 1H), 7.28–7.25 (m, 1H). LC-MS(ESI):m / z:383.3[M+H] + .

[0108] Example 7

[0109] This example provides a heterocyclic compound I-7, whose reaction equation and preparation method are as follows:

[0110] Step 1: Dissolve methyl 6-aminoindole-4-carboxylate (190.2 mg, 1.0 mmol, 1.0 equivalent), cuprous iodide (190.0 mg, 1.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (88.2 mg, 1.0 mmol, 1.0 equivalent), potassium carbonate (276.0 mg, 2.0 mmol, 2.0 equivalent), and 4-iodotrifluorotoluene (272.0 mg, 1.0 mmol, 1.0 equivalent) in 3 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was complete as monitored by HPLC-MS, 6 mL of aqueous solution was added to quench the reaction, followed by extraction with 18 mL of ethyl acetate three times. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound 7b was obtained by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 50%), yielding 168.3 mg of white solid compound 7b, with a yield of 50.2%. LC-MS (ESI): m / z 335.2 [M+H] + .

[0111] Step 2: Dissolve 7b (168.3 mg, 0.5 mmol, 1.0 equivalent) in 3 mL of dichloromethane, then add triethylamine (150.0 mg, 1.5 mmol, 3.0 equivalent). Slowly add acryloyl chloride (45.7 mg, 0.5 mmol) under ice bath conditions. After stirring at room temperature for 6 hours, monitor the reaction for completeness by HPLC-MS, then quench the reaction with water. Extract three times with DCM, dry with anhydrous sodium sulfate, concentrate under vacuum, and separate and purify by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 30%) to obtain 155.7 mg of white solid compound 7c, yield 95%. 1 H NMR (500MHz, DMSO-d6) δ7.94(d,J=8.3Hz,2H),7.79(d,J=8.3Hz,2H),7.54(d,J=3.3Hz,1H),7.29(d,J =1.9Hz,1H),7.11(d,J=1.1Hz,1H),7.02(d,J=4.1Hz,1H),5.26(s,2H),3.90(s,3H).LC-MS(ESI):m / z 389.2[M+H] + .

[0112] Step 3: Dissolve 7c (155.7 mg, 0.40 mmol, 1.0 equivalent) in 5 mL of ethanol, then add potassium hydroxide aqueous solution (0.04 mL, 0.40 mmol, 10 N, 1.0 equivalent). Stir overnight at room temperature. After the reaction is complete as monitored by HPLC-MS, the solvent is removed by vacuum concentration. The pH of the solution is adjusted to 1 with 6 N hydrochloric acid, and a precipitate forms. The solvent is removed by filtration, and the residue is dried to obtain 142.6 mg of white solid compound 7d, with a yield of 95.0%. No purification is required; it can be used directly in the next reaction. LC-MS (ESI): m / z 375.2 [M+H] + .

[0113] Step 4: Dissolve compound B-1 (153.3 mg, 0.5 mmol, 1.3 equivalents) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. N,N-dimethylformamide was added, followed by 7d (142.6 mg, 0.38 mmol, 1.0 equivalent). The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (144.4 mg, 0.38 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to be complete, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 30%) to give 20.0 mg of white solid compound I-7, with a yield of 9.4%.

[0114] The NMR data of the product are as follows:

[0115] 1H NMR (500MHz, CDCl3) δ8.52(s,1H),8.33(s,1H),7.78(d,J=8.2Hz,2H),7.60(d,J=8.2 Hz,2H),7.38(dd,J=5.5,2.0Hz,3H),7.30(d,J=7.4Hz,2H),7.15(d,J=1.8Hz,1H),7. 06(t,J=7.2Hz,1H),6.84(s,1H),6.64(d,J=3.3Hz,1H),6.42(d,J=18.3Hz,1H),6.29 (dd,J=16.9,10.1Hz,1H),5.72(d,J=11.5Hz,1H),3.93–3.39(m,8H).LC-MS(ESI):m / z 562.1[M+H] + .

[0116] Example 8

[0117] This example provides a heterocyclic compound I-8, whose reaction equation and preparation method are as follows:

[0118] Step 1: Indole-4-carboxylic acid methyl ester (175.0 mg, 1.0 mmol, 1.0 equivalent), potassium hydroxide (56.1 mg, 1.0 mmol, 1.0 equivalent), and 4-trifluoromethylbenzyl bromide (239.0 mg, 1.0 mmol, 1.0 equivalent) were dissolved in 3 mL of N,N-dimethylformamide. The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by HPLC-MS, the reaction was quenched with 6 mL of aqueous solution and extracted three times with 18 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound was then separated and purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 60%) to give 161.6 mg of white solid compound 8b, yield 50.5%. LC-MS (ESI): m / z: 320.2 [M+H] + .

[0119] Step 2: Dissolve compound B-1 (153.3 mg, 0.5 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following N,N-dimethylformamide, 8b (161.6 mg, 0.50 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (190.0 mg, 0.50 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 30%) to give 47.5 mg of white solid compound I-8, with a yield of 18.8%.

[0120] The NMR data of the product are as follows:

[0121] 1 H NMR(500MHz, CDCl3)δ7.58(d,J=8.0Hz,2H),7.39–7.29(m,4H),7.24–7.16(m,5H),7.06(t,J=7.3Hz,1H), 6.72(s,1H),6.63–6.54(m,1H),5.42(s,2H),3.91(s,2H),3.68–3.35(m,6H).LC-MS(ESI):m / z507.5[M+H] + .

[0122] Example 9

[0123] This example provides a heterocyclic compound I-9, whose reaction equation and preparation method are as follows:

[0124] Step 1: Indole-4-carboxylic acid methyl ester (175.0 mg, 1.0 mmol, 1.0 equivalent), potassium hydroxide (56.1 mg, 1.0 mmol, 1.0 equivalent), and 4-trifluoromethoxybenzyl bromide (255.0 mg, 1.0 mmol, 1.0 equivalent) were dissolved in 3 mL of N,N-dimethylformamide. The mixture was stirred overnight at room temperature. After the reaction was monitored by HPLC-MS to ensure complete reaction, 6 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 18 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound 9b was then separated and purified by a Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 60%) to obtain 208.8 mg of white solid compound 9b, with a yield of 62.1%. 1 HNMR(500MHz, CDCl3)δ8.07(d,J=7.4Hz,1H),7.53(d,J=8.1Hz,1H),7.34(s,2H),7.31– 7.28(m,1H),7.18(d,J=8.5Hz,2H),7.13(d,J=8.5Hz,2H),5.42(s,2H).LC-MS(ESI):m / z 336.2[M+H] + .

[0125] Step 2: Dissolve compound B-1 (214.2 mg, 0.7 mmol, 1.1 equivalents) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following N,N-dimethylformamide, 9b (208.8 mg, 0.62 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (235.6 mg, 0.62 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 30%) to obtain 83.5 mg of white solid compound I-9, with a yield of 25.7%.

[0126] The NMR data of the product are as follows:

[0127] 1H NMR (500MHz, CDCl3) δ7.42–7.29(m,4H),7.25–7.20(m,2H),7.19–7.10(m,5H),7.05(t,J=7.4 Hz,1H),6.72(s,1H),6.57(d,J=3.3Hz,1H),5.37(s,2H),3.98–3.39(m,8H).LC-MS(ESI):m / z 523.2[M+H] + .

[0128] Example 10

[0129] This example provides a heterocyclic compound I-10, the reaction equation and preparation method of which are as follows:

[0130] Step 1: Indole-4-carboxylic acid methyl ester (175.0 mg, 1.0 mmol, 1.0 equivalent), potassium hydroxide (56.1 mg, 1.0 mmol, 1.0 equivalent), and 2-cyclohexylbromoethane (191.1 mg, 1.0 mmol, 1.0 equivalent) were dissolved in 3 mL of N,N-dimethylformamide. The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by HPLC-MS, the reaction was quenched with 6 mL of aqueous solution and extracted three times with 18 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound was then separated and purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 60%) to give 145.7 mg of white solid compound 10b, yield 53.5%. LC-MS (ESI): m / z 272.3 [M+H] + .

[0131] Step 2: Dissolve compound B-1 (153.0 mg, 0.5 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following N,N-dimethylformamide, 9b (145.7 mg, 0.54 mmol, 1.1 equivalents) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (190.0 mg, 0.5 mmol, 1.0 equivalents) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 30%) to obtain 92.5 mg of white solid compound I-10, with a yield of 40.3%.

[0132] The NMR data of the product are as follows:

[0133] 1 H NMR (500MHz, CDCl3) δ7.42(d,J=8.3Hz,1H),7.38–7.35(m,2H),7.34–7.30(m,2H),7.27–7.22(m,1H),7.18–7.15(m,1H),7.08(t,J=7.3Hz,1H),6.4 9(d,J=3.1Hz,1H),6.38(s,1H),4.18(t,J=7.6Hz,2H),4.02–3.44(m,8H), 1.77–1.75(m,2H),1.31–1.14(m,9H),1.05–0.97(m,2H).LC-MS(ESI):m / z 459.3[M+H] + .

[0134] Example 11

[0135] This example provides a heterocyclic compound I-11, whose reaction equation and preparation method are as follows:

[0136] Step 1: Dissolve intermediate 1c (642.0 mg, 2.0 mmol, 1.0 equivalent) and 1-tert-butyloxycarbonylpiperazine (372.5 mg, 2.0 mmol, 1.0 equivalent) in 10 mL Following N,N-dimethylformamide, N,N-diisopropylethylamine (260.0 mg, 2.00 mmol, 2.0 equivalent) was added and stirred at room temperature for 15 minutes. Then, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (760.0 mg, 2.00 mmol, 1.0 equivalent) was added and the mixture was stirred at room temperature overnight. After the reaction was monitored by HPLC-MS to ensure complete reaction, 20 mL of aqueous solution was added to quench the reaction, followed by three fractions extraction with 60 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 30%) to give 566.1 mg of white solid compound 11b, yield 59.8%. 1 H NMR (500MHz, CDCl3) δ7.57–7.51(m,2H),7.43–7.34(m,3H),7.23–7.18(m,1H),7.03–6.97(m,1 H),6.95–6.92(m,1H),6.46(d,J=3.4Hz,1H),3.63–3.09(m,8H),2.57(s,9H).LC-MS(ESI):m / z 474.2[M+H] + .

[0137] Step 2: Dissolve intermediate 11b (95.0 mg, 0.20 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 1.0 mL of trifluoroacetic acid. Stir at room temperature for 3 hours. After HPLC-MS monitoring to ensure complete removal of the tert-butyloxycarbonyl group from the reactant 11b, concentrate under vacuum to remove the solvent. Dissolve the reaction residue in 3 mL of N,N-dimethylformamide, then adjust the pH of the solution to alkaline with N,N-diisopropylethylamine. Add cyclohexyl isocyanate (27.6 mg, 0.22 mmol, 1.1 equivalent), stir at room temperature overnight. After HPLC-MS monitoring to ensure complete reaction, quench the reaction with 5 mL of aqueous solution, then extract three times with 18 mL of ethyl acetate. Combine the organic phases, wash with saturated sodium chloride aqueous solution and water, dry with anhydrous sodium sulfate, and concentrate under vacuum. Separate and purify by preparative liquid chromatography to obtain 86.4 mg of white solid compound I-11, yield 86.7%.

[0138] The NMR data of the product are as follows:

[0139] 1H NMR(500MHz, CDCl3)δ7.81(d,J=8.3Hz,2H),7.65–7.60(m,3H),7.41(d,J=3.3Hz, 1H),7.29(d,J=7.6Hz,1H),7.21(d,J=7.2Hz,1H),6.73(d,J=3.3Hz,1H),4.29(d,J =7.6Hz,1H),4.00–3.78(m,2H),3.68–3.63(m,1H),3.51–3.33(m,5H),1.98–1.90 (m,2H),1.75–1.67(m,2H),1.45–1.30(m,3H),1.18–1.06(m,3H).LC-MS(ESI):m / z 499.2 [M+H] + .

[0140] Example 12

[0141] This example provides a heterocyclic compound I-12, the reaction equation and preparation method of which are as follows:

[0142] Step 1: Dissolve intermediate 11b (95.0 mg, 0.20 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 1.0 mL of trifluoroacetic acid. Stir at room temperature for 3 hours. After HPLC-MS monitoring to ensure complete removal of the tert-butyloxycarbonyl group from the reactant 11b, concentrate under vacuum to remove the solvent. Dissolve the reaction residue in 3 mL of N,N-dimethylformamide, then adjust the pH of the solution to alkaline with N,N-diisopropylethylamine. Add 3-carboxymethoxyphenyl isocyanate (39.0 mg, 0.22 mmol, 1.1 equivalent), stir at room temperature overnight. After HPLC-MS monitoring to ensure complete reaction, quench the reaction with 5 mL of aqueous solution, then extract three times with 18 mL of ethyl acetate. Combine the organic phases, wash with saturated sodium chloride aqueous solution and water, dry with anhydrous sodium sulfate, and concentrate under vacuum. Separate and purify by preparative liquid chromatography to obtain 94.0 mg of pale yellow solid I-12, yield 85.4%.

[0143] The NMR data of the product are as follows:

[0144] 1H NMR (500MHz, CDCl3) δ8.54 (d, J=9.8Hz, 1H), 8.01 (dd, J=8.1, 1.7Hz, 1H), 7.82 (d, J=8.3Hz,2H),7.64(dd,J=8.3,5.1Hz,3H),7.55–7.49(m,1H),7.42(d,J=3.3Hz,1H ),7.33–7.27(m,1H),7.25(d,J=6.7Hz,1H),7.00(t,J=7.1Hz,1H),6.75(d,J=3.3 Hz,1H),4.09–3.91(m,2H),3.85–3.50(m,6H),1.29–1.24(m,3H).LC-MS(ESI):m / z 551.2[M+H] + .

[0145] Example 13

[0146] This example provides a heterocyclic compound I-13, the reaction equation and preparation method of which are as follows:

[0147] Step 1: Dissolve intermediate 11b (95.0 mg, 0.20 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 1.0 mL of trifluoroacetic acid. Stir at room temperature for 3 hours. After complete removal of the tert-butyloxycarbonyl group of the reactant 11b by HPLC-MS, concentrate under vacuum to remove the solvent. Dissolve the reaction residue in 3 mL of N,N-dimethylformamide, then adjust the pH of the solution to alkaline with N,N-diisopropylethylamine. Add 2-naphthyl isocyanate (37.2 mg, 0.22 mmol, 1.1 equivalent), stir at room temperature overnight. After the reaction is complete by HPLC-MS, quench the reaction with 5 mL of aqueous solution, then extract three times with 18 mL of ethyl acetate. Combine the organic phases, wash with saturated sodium chloride aqueous solution and water, dry with anhydrous sodium sulfate, and concentrate under vacuum. Separate and purify by preparative liquid chromatography to obtain 60.6 mg of white solid compound I-13, yield 55.8%.

[0148] The NMR data of the product are as follows:

[0149] 1H NMR (500MHz, CDCl3) δ7.93(d,J=2.3Hz,1H),7.82(d,J=8.4Hz,2H),7.81–7.72(m,4H),7.64(d,J=8.3Hz,3H),7.45–7.42(m,2H),7.41–7.39( m,1H),7.33–7.29(m,1H),7.26–7.23(m,1H),6.76(dd,J=3.4,0.8Hz,1H),4.06–3.95(m,2H),3.73–3.52(m,6H).LC-MS(ESI):m / z:543.2(M+H) + LC-MS(ESI): m / z 543.2 [M+H] + .

[0150] Example 14

[0151] This example provides a heterocyclic compound I-14, whose reaction equation and preparation method are as follows:

[0152] Step 1: Dissolve intermediate 11b (95.0 mg, 0.20 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 1.0 mL of trifluoroacetic acid. Stir at room temperature for 3 hours. After complete removal of the tert-butoxycarbonyl group from the reactant 11b by HPLC-MS, concentrate under vacuum to remove the solvent. Dissolve the reaction residue in 3 mL of N,N-dimethylformamide, then adjust the pH of the solution to alkaline with N,N-diisopropylethylamine. Add 1-isocyanate quaternane (39.0 mg, 0.22 mmol, 1.1 equivalent), stir at room temperature overnight. After the reaction is complete by HPLC-MS, quench the reaction with 5 mL of aqueous solution, then extract three times with 18 mL of ethyl acetate. Combine the organic phases, wash with saturated sodium chloride aqueous solution and water, dry with anhydrous sodium sulfate, and concentrate under vacuum. Separate and purify by preparative liquid chromatography to obtain 72.1 mg of pale yellow solid compound I-14, yield 65.5%.

[0153] The NMR data of the product are as follows:

[0154] 1H NMR (500MHz, CDCl3) δ7.93(d,J=2.3Hz,1H),7.82(d,J=8.4Hz,2H),7.81–7.72(m,4H),7.64(d,J=8.3Hz,3H),7.45–7.42(m,2H),7.41–7.39(m ,1H),7.33–7.29(m,1H),7.26–7.23(m,1H),6.76(dd,J=3.4,0.8Hz,1H),4.06–3.95(m,2H),3.73–3.52(m,6H).LC-MS(ESI):m / z:551.2[M+H] + .

[0155] Example 15

[0156] This example provides a heterocyclic compound I-15, the reaction equation and preparation method of which are as follows:

[0157] Step 1: Dissolve intermediate 11b (95.0 mg, 0.20 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 1.0 mL of trifluoroacetic acid. Stir at room temperature for 3 hours. After complete removal of the tert-butyloxycarbonyl group from the reactant 11b by HPLC-MS, concentrate under vacuum to remove the solvent. Dissolve the reaction residue in 3 mL of N,N-dimethylformamide, then adjust the pH of the solution to alkaline with N,N-diisopropylethylamine. Add phenyl thioisocyanate (29.7 mg, 0.22 mmol, 1.1 equivalent), stir at room temperature overnight. After the reaction is complete by HPLC-MS, quench the reaction with 5 mL of aqueous solution, then extract three times with 18 mL of ethyl acetate. Combine the organic phases, wash with saturated sodium chloride aqueous solution and water, dry with anhydrous sodium sulfate, and concentrate under vacuum. Separate and purify by preparative liquid chromatography to obtain 52.6 mg of pale yellow solid compound I-15, yield 51.7%.

[0158] The NMR data of the product are as follows:

[0159] 1 H NMR (500MHz, CDCl3) δ7.81(d,J=8.3Hz,2H),7.62(d,J=8.4Hz,3H),7.42(d,J=3.4Hz,1H),7.37–7.32(m,3H),7.30–7.27(m,1H ),7.17(t,J=7.4Hz,1H),7.13(d,J=8.6Hz,2H),6.72(d,J=3.3Hz,1H),4.00–3.76(m,6H),3.64–3.39(m,2H).LC-MS(ESI):m / z 509.1[M+H] + .

[0160] Example 16

[0161] This example provides a heterocyclic compound I-16, whose reaction equation and preparation method are as follows:

[0162] Step 1: Dissolve indazole-4-carboxylic acid (162.2 mg, 1.0 mmol, 1.0 equivalent), cuprous iodide (190.0 mg, 1.0 mmol, 1.0 equivalent), N,N-dimethylethylenediamine (88.2 mg, 1.0 mmol, 1.0 equivalent), potassium carbonate (276.0 mg, 2.0 mmol, 2.0 equivalent), and 1-iodo-4-(trifluoromethoxy)benzene (288.0 mg, 1.0 mmol, 1.0 equivalent) in 3 mL. In N,N-dimethylformamide, the reaction was carried out under argon protection at 140°C with stirring for 6 hours. After the reaction was complete as monitored by HPLC-MS, 6 mL of aqueous solution was added to quench the reaction, followed by extraction with 18 mL of ethyl acetate three times. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The purified compound was then separated and purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 50%) to give 61.2 mg of yellow solid compound 16b, with a yield of 20.0%. LC-MS (ESI): m / z 306.2 [M+H] + .

[0163] Step 2: Dissolve compound B-1 (61.2 mg, 0.20 mmol, 1.0 equivalent) in 2 mL of dichloromethane, then add 0.5 mL of trifluoroacetic acid. Stir at room temperature for 2 hours. Monitor the complete conversion of the starting material compound B-1 to N-phenylpiperazine-1-carboxamide by HPLC-MS. After vacuum concentration to remove the solvent, dissolve the reaction residue in 3 mL of dichloromethane. Following N,N-dimethylformamide, 9b (61.2 mg, 0.20 mmol, 1.0 equivalent) was added. The pH of the solution was then adjusted to alkaline with N,N-diisopropylethylamine. Finally, HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (76.0 mg, 0.2 mmol, 1.0 equivalent) was added. The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to ensure complete reaction, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 50%) to give 21.0 mg of white solid compound I-16, with a yield of 21.2%.

[0164] The NMR data of the product are as follows:

[0165] 1 H NMR(500MHz, CDCl3)δ8.11–8.06(m,1H),7.90(t,J=9.8Hz,2H),7.86–7.79(m,2H),7.41–7 .26(m,6H),7.10–7.06(m,1H),6.54(d,J=4.6Hz,1H),4.00–3.49(m,8H).LC-MS(ESI):m / z 494.2[M+H] + .

[0166] Example 17

[0167] This example provides a heterocyclic compound I-17, whose reaction equation and preparation method are as follows:

[0168] Step 1: Intermediate I-3 (104.0 mg, 0.20 mmol, 1.0 equivalent) and starting material (S)-1-phenylethylamine (24.2 mg, 0.20 mmol, 1.0 equivalent) were dissolved in 2.0 mL of methanol. The solution was heated to 50 °C, and then 2.0 mL of tetrahydrofuran was added. After stirring the mixture for 1 hour, sodium cyanoborohydride (18.8 mg, 0.30 mmol, 1.5 equivalent) was added. After cooling to room temperature, the mixture was stirred for 4 hours. After the reaction was monitored by HPLC-MS to be complete, 2.0 mL of 1N hydrochloric acid solution was added to the reaction solution and stirred at room temperature for 1 hour. Then, the pH was adjusted to 8 with 1N sodium hydroxide solution, and the reaction solution was diluted with 10.0 mL of water. The mixture was extracted once with 10.0 mL of dichloromethane. The organic phase was washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The mixture was separated and purified by preparative liquid chromatography to obtain 42.0 mg of white solid compound I-17, with a yield of 33.6%.

[0169] The NMR and mass spectrometry data of the product are as follows:

[0170] 1 H NMR (500MHz, CDCl3) δ7.82–7.75(m,2H),7.63–7.55(m,3H),7.49(s,3H),7.41–7.33(m,4H),7.30–7.24(m,3H),7.16–7.12(m,2H),7.12–7.09( m,1H),6.96–6.91(m,1H),4.69–4.46(m,1H),4.34–4.26(m,1H),4.22–3.92(m,3H),3.88–3.41(m,7H),1.70(d,J=6.7Hz,3H).LC-MS(ESI):m / z 626.1(M+H + ).

[0171] Example 18

[0172] This example provides a heterocyclic compound I-18, whose reaction equation and preparation method are as follows:

[0173] Following a similar process to Example 17, 25.9 mg of the title compound was prepared from intermediate I-3 and starting material (S)-1-(2-pyridine)ethylamine, as a white solid, in a yield of 20.7%.

[0174] The NMR and mass spectrometry data of the product are as follows:

[0175] 1 H NMR (500MHz, CDCl3) δ8.49 (s, 1H), 7.81 (d, J = 8.2Hz, 2H), 7.74–7.67 (m, 1H), 7. 63–7.55(m,3H),7.51(s,1H),7.42–7.35(m,3H),7.33–7.29(m,1H),7.26–7.17 (m,3H),7.16–7.09(m,2H),6.97(t,J=7.4Hz,1H),4.69–4.46(m,1H),4.38–4.1 1(m,3H),3.86–3.17(m,8H),1.66(d,J=6.8Hz,3H).LC-MS(ESI):m / z627.0[M+H] + .

[0176] Example 19

[0177] This example provides a heterocyclic compound I-19, whose reaction equation and preparation method are as follows:

[0178] Following a similar process to Example 17, 33.0 mg of the title compound was prepared from intermediate I-3 and starting material 1-thiomorpholine oxide as a white solid, with a yield of 26.4%.

[0179] The NMR and mass spectrometry data of the product are as follows:

[0180] 1H NMR (500MHz, CDCl3) δ7.88–7.77(m,2H),7.70–7.59(m,3H),7.41–7.35(m,2H),7.32(t,J=7.8Hz,3H),7. 26–7.11(m,3H),7.00(s,1H),4.70–4.20(m,2H),4.00–3.46(m,8H),3.40–2.50(m,8H).LC-MS(ESI):m / z 624.1[M+H] + .

[0181] Example 20

[0182] This example provides a heterocyclic compound I-20, whose reaction equation and preparation method are as follows:

[0183] Following a similar process to Example 17, 33.0 mg of the title compound was prepared from intermediate I-3 and the starting material thiomorpholine-1,1-dioxide, as a white solid, in a yield of 26.4%.

[0184] The NMR and mass spectrometry data of the product are as follows:

[0185] 1 H NMR (500MHz, CDCl3) δ8.02(s,1H),7.89(d,J=8.3Hz,2H),7.67(d,J=8.3Hz,2H),7.53(d,J=7.4Hz,1H),7.45–7.40(m,1H),7.3 4(d,J=7.4Hz,2H),7.30(s,1H),7.26–7.21(m,2H),7.03–6.99(m,2H),4.55–2.94(m,12H),2.31–0.65(m,4H).LC-MS(ESI):m / z 640.2[M+H] +

[0186] Example 21

[0187] This example provides a heterocyclic compound I-21, whose reaction equation and preparation method are as follows:

[0188] Step 1: Intermediate I-3 (520.0 mg, 1.00 mmol, 1.0 equivalent) and starting material hydroxylamine hydrochloride (210.0 mg, 3.0 mmol, 3.0 equivalent) were dissolved in 10.0 mL of pyridine. After stirring the mixture for 8 hours, 25.0 mL of 2N hydrochloric acid solution was added, and the mixture was stirred at room temperature for 1 hour. Subsequently, the mixture was extracted three times with 45.0 mL of ethyl acetate solution. The combined organic phases were washed with 2N hydrochloric acid solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain 482.4 mg of crude product 21b (LC-MS (ESI): m / z 536.1 [M+H)). + (Yield 90.0%), it can be used in the next reaction without purification.

[0189] Step 1: The crude intermediate 21b (482.4 mg, 0.90 mmol, 1.0 equivalent) and the starting zinc powder (588.6 mg, 9.0 mmol, 10.0 equivalent) were dissolved in 20 mL of acetic acid. The mixture was stirred at 50 °C for 10 hours. The remaining zinc powder was removed with diatomaceous earth. The reaction solution was slowly added dropwise to 60 mL of 2 M sodium hydroxide solution. The mixture was then extracted three times with 60 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The solution was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 75%) to obtain 353.4 mg of white solid compound I-21, with a yield of 75.2%.

[0190] The NMR and mass spectrometry data of the product are as follows:

[0191] 1 H NMR(500MHz, CDCl3)δ8.40(s,1H),7.64–7.59(m,2H),7.54(s,1H),7.49–7.42(m,3H),7.37–7.31(m,2H),7.13(t,J=7.8Hz ,1H),7.04(t,J=7.8Hz,2H),6.92–6.88(m,1H),6.84–6.78(m,1H),4.26–4.01(m,2H),3.67–3.15(m,8H).LC-MS(ESI):m / z 522.1[M+H] + .

[0192] Example 22

[0193] This example provides a heterocyclic compound I-22, whose reaction equation and preparation method are as follows:

[0194] Step 1: Dissolve intermediate I-21 (52.2 mg, 0.10 mmol, 1.0 equivalent) in 3 mL Following N,N-dimethylformamide, 2-fluoroacrylic acid (13.5 mg, 0.15 mmol, 1.5 equivalents) was added, followed by N,N-diisopropylethylamine (19.5 mg, 0.15 mmol, 1.5 equivalents) and HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (38.0 mg, 0.1 mmol, 1.0 equivalents). The mixture was stirred at room temperature for 6 hours. After the reaction was monitored by HPLC-MS to be complete, the reaction was quenched with 5 mL of aqueous solution and extracted three times with 30 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% to ethyl acetate / petroleum ether = 80%) to give 22.0 mg of white solid compound I-21, yield 37.0%.

[0195] The NMR and mass spectrometry data of the product are as follows:

[0196] 1 H NMR(500MHz, CDCl3)δ7.84–7.80(m,3H),7.64–7.59(m,3H),7.50(s,1H),7.39–7.35(m,2H),7.32–7.28(m,3H),7.12–7.09(m,1H),7.08 –7.04(m,1H),6.72(s,1H),5.65(dd,J=47.1,3.2Hz,1H),5.11(dd,J=15.0,3.1Hz,1H),3.98(s,2H),3.75–3.30(m,8H).LC-MS(ESI):m / z 594.1[M+H] + .

[0197] Example 23

[0198] This example provides a heterocyclic compound I-23, whose reaction equation and preparation method are as follows:

[0199] Following a similar process to Example 22, 29.5 mg of the title compound was prepared from intermediate I-3 and starting material acrylic acid, as a white solid, in a yield of 51.3%.

[0200] The NMR and mass spectrometry data of the product are as follows:

[0201] 1H NMR (500MHz, CDCl3) δ7.85–7.79(m,2H),7.65–7.59(m,3H),7.53(s,1H),7.41–7.30(m,6H),7.13–7.07(m,2H),6.50(s,1H),6.29(dd,J =17.0,1.5Hz,1H),6.14(dd,J=17.0,10.3Hz,1H),5.63(dd,J=10.3,1.5Hz,1H),4.12–4.01(m,2H),3.79–3.45(m,8H).LC-MS(ESI):m / z 576.1[M+H] + .

[0202] Example 24

[0203] This example provides a heterocyclic compound I-24, whose reaction equation and preparation method are as follows:

[0204] Step 1: Intermediate I-21 (52.2 mg, 0.10 mmol, 1.0 equivalent) was dissolved in 5 mL of ultra-dry dichloromethane, followed by the addition of triethylamine (60.6 mg, 0.6 mmol, 6 equivalent). Then, 2-chloroethanesulfonyl chloride (16.3 mg, 0.10 mmol, 1.0 equivalent) was slowly added dropwise under ice bath conditions. After the addition was complete, the mixture was stirred overnight at room temperature. After the reaction was monitored by HPLC-MS to be complete, the reaction was quenched with 5 mL of aqueous solution and extracted three times with 15 mL of dichloromethane. The organic phases were combined, washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by Flash column chromatography (gradient: ethyl acetate / petroleum ether = 5% ~ ethyl acetate / petroleum ether = 85%) to obtain 15.6 mg of white solid compound I-24, with a yield of 25.5%.

[0205] The NMR and mass spectrometry data of the product are as follows:

[0206] 1 H NMR(500MHz, CDCl3)δ7.86–7.80(m,2H),7.69–7.50(m,4H),7.43(s,1H),7.39–7.36(m,1H),7.32–7.18(m,4H),7.12–7.0 0(m,2H),6.50–6.41(m,1H),6.34–6.07(m,2H),5.89–5.79(m,1H),4.50–4.09(m,2H),3.75–3.20(m,8H).LC-MS(ESI):m / z 612.2[M+H] + .

[0207] Example 25

[0208] This example provides a heterocyclic compound I-25, whose reaction equation and preparation method are as follows:

[0209] Step 1: Intermediate I-21 (260.0 mg, 0.50 mmol, 1.0 equivalent) was dissolved in 3 mL of ultra-dry dichloromethane, followed by the addition of triethylamine (151.5 mg, 1.5 mmol, 3.0 equivalent) and triphosgene (147.9 mg, 0.5 mmol, 1.0 equivalent). After stirring at room temperature for 15 minutes, (S)-1-(2-pyridine)ethylamine (61.1 mg, 0.50 mmol, 1.0 equivalent) was added, and triethylamine (151.5 mg, 1.5 mmol, 3.0 equivalent) was added as well. The reaction solution was reacted at room temperature for 16 hours, then diluted with 20 mL of dichloromethane, washed with saturated ammonium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The solution was then separated and purified by preparative liquid chromatography to obtain 38.0 mg of white solid compound I-25, with a yield of 11.3%.

[0210] The NMR and mass spectrometry data of the product are as follows:

[0211] 1 H NMR(500MHz,DMSO-d6)δ8.75–8.43(m,2H),7.96(d,J=7.9Hz,2H),7.85–7.61( m,5H),7.44(m,3H),7.26(m,4H),7.12(d,J=7.1Hz,1H),6.93(t,J=5.0Hz,1H), 6.71(s,1H),6.41(d,J=6.4Hz,1H),4.85(t,J=5.0Hz,1H),4.46–4.13(m,2H),3 .80–3.57(m,5H),3.26(m,3H),1.37–1.27(m,3H).LC-MS(ESI):m / z670.1[M+H] + .

[0212] Example 26

[0213] This example provides a heterocyclic compound I-26, whose reaction equation and preparation method are as follows:

[0214] Following a similar process to Example 25, 22.0 mg of the title compound was prepared from intermediate I-21 and starting material (S)-1-phenylethylamine as a white solid, with a yield of 10.5%.

[0215] The NMR and mass spectrometry data of the product are as follows:

[0216] 1H NMR(500MHz,DMSO-d6)δ8.59(s,1H),7.96(d,J=8.0Hz,2H),7.79(s,2H),7.70(m,1 H),7.57(d,J=7.4Hz,1H),7.45(d,J=7.4Hz,2H),7.27(m,7H),7.15(m,2H),6.94(t, J=5.0Hz,1H),6.41(m,1H),5.99(d,J=5.9Hz,1H),4.74(t,J=5.0Hz,1H),4.40(m,1H ),4.13(m,1H),3.69(m,5H),3.50–3.40(m,3H),1.40–1.26(m,4H).LC-MS(ESI):m / z 669.3[M+H] + .

[0217] Example 27

[0218] This example provides a heterocyclic compound I-27, the reaction equation and preparation method of which are as follows:

[0219] Following a similar process to Example 25, 20.0 mg of the title compound was prepared from intermediate I-21 and starting material phenethylamine, as a white solid, in a yield of 6.1%.

[0220] The NMR and mass spectrometry data of the product are as follows:

[0221] 1 H NMR(500MHz,DMSO-d6)δ8.61(s,1H),7.98–7.95(m,2H),7.81(m,2H),7.74–7. 70(m,1H),7.62(m,1H),7.46(d,J=7.4Hz,2H),7.31(m,1H),7.26–7.22(m,6H), 7.20–7.17(m,1H),7.14–7.10(m,1H),6.94(t,J=5.0Hz,1H),6.48–6.44(m,1H) ,6.13(t,J=5.0Hz,1H),4.28–4.16(m,4H),3.79–3.44(m,8H).LC-MS(ESI):m / z 655.1[M+H] + .

[0222] Example 28

[0223] This example provides a heterocyclic compound I-28, whose reaction equation and preparation method are as follows:

[0224] Following a similar process to Example 25, 21.0 mg of the title compound was prepared from intermediate I-21 and starting material 1-thiomorpholine oxide as a white solid, with a yield of 6.3%.

[0225] The NMR and mass spectrometry data of the product are as follows:

[0226] 1 H NMR (500MHz, DMSO-d6) δ8.61(s,1H),7.96(d,J=7.9Hz,2H),7.83(d,J=7.8Hz,2H),7.71(d,J =7.7Hz,1H),7.66(s,1H),7.44(d,J=7.4Hz,2H),7.31(t,J=10.0Hz,1H),7.23(t,J=10.0Hz, 2H),7.11(m,1H),6.94(t,J=10.0Hz,1H),6.88(t,J=5.0Hz,1H),4.38(m,2H),3.80(m,4H),3 .73–3.55(m,5H),3.41(m,2H),3.24(s,1H),2.91–2.80(m,2H),2.62(m,2H).LC-MS(ESI):m / z 667.6[M+H] + .

[0227] Example 29

[0228] This example provides a heterocyclic compound I-29, whose reaction equation and preparation method are as follows:

[0229] Following a similar process to Example 25, 50.0 mg of the title compound was prepared from intermediate I-21 and starting material thiomorpholine-1,1-dioxide, as a white solid, in a yield of 14.6%.

[0230] The NMR and mass spectrometry data of the product are as follows:

[0231] 1H NMR(500MHz,DMSO-d6)δ8.86(s,1H),7.95(d,J=8.3Hz,2H),7.84(d,J=8.3Hz, 2H),7.73–7.67(m,2H),7.53–7.47(m,2H),7.33–7.29(m,1H),7.24–7.19(m,2 H),7.17–7.13(m,1H),7.10(d,J=7.2Hz,1H),6.92(t,J=7.2Hz,1H),4.38(s,2 H),3.88–3.62(m,8H),3.24–3.10(m,4H),1.35–1.25(m,4H).LC-MS(ESI):m / z 683.1[M+H] + .

[0232] Example 30

[0233] This example provides a heterocyclic compound I-30, the reaction equation and preparation method of which are as follows:

[0234] Step 1: Intermediate I-3 (522.0 mg, 1.00 mmol, 1.0 equivalent) was dissolved in a mixed solution of tert-butanol (15 mL) and 2-methyl-2-butene (3 mL), followed by the addition of sodium chlorite (1.06 g, 11.7 mmol, 11.7 equivalent) and sodium dihydrogen phosphate (1.06 g, 8.8 mmol, 8.8 equivalent) in water (6 mL). The reaction mixture was stirred at room temperature for 48 hours. The mixture was diluted with water and extracted three times with 30 mL of ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate and concentrated under vacuum. The purified compound was obtained by preparative liquid chromatography (HPLC) to yield 402.7 mg of a white solid, compound I-30, in 75.5% yield.

[0235] The NMR and mass spectrometry data of the product are as follows:

[0236] 1 H NMR(500MHz, CDCl3)δ10.06(s,1H),8.20–8.13(m,1H),8.05(s,1H),7.95–7.88(m,3H), 7.72–7.66(m,4H),7.42–7.35(m,4H),6.47(s,1H),3.55–3.33(m,8H).LC-MS(ESI):m / z 537.1[M+H] + .

[0237] Example 31

[0238] This example provides a heterocyclic compound I-31, the reaction equation and preparation method of which are as follows:

[0239] Step 1: Dissolve intermediate I-30 (53.7 mg, 0.10 mmol, 1.0 equivalent) and starting material (S)-1-(2-pyridine)ethylamine (12.0 mg, 0.10 mmol, 1.0 equivalent) in 2 mL of N,N-dimethylformamide solution, then add N,N-diisopropylethylamine (26.0 mg, 0.20 mmol, 2.0 equivalent) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl) Urea hexafluorophosphate (57.0 mg, 0.15 mmol, 1.5 equivalents) was reacted with the solution at room temperature for 2 hours. After the reaction was complete as monitored by HPLC-MS, 5 mL of aqueous solution was added to quench the reaction, and then the mixture was extracted three times with 15 mL of ethyl acetate. The combined organic phases were washed with saturated sodium chloride aqueous solution and water, dried with anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then separated and purified by preparative liquid chromatography to obtain 18.8 mg of pale yellow solid compound I-31, with a yield of 29.3%.

[0240] The NMR and mass spectrometry data of the product are as follows:

[0241] 1 H NMR (500MHz, DMSO-d6) δ8.60–8.34(m,4H),8.03(d,J=8.3Hz,2H),7.94(d,J=8.3Hz,2H),7.79–7.64(m,2H),7.49–7.42(m,2H),7.39–7.17(m ,4H),7.15–7.02(m,2H),6.99–6.87(m,1H),5.19–5.06(m,1H),3.70–3.42(m,5H),3.29–2.90(m,4H),1.50(d,J=7.0Hz,3H).LC-MS(ESI):m / z 641.2[M+H] + .

[0242] Example 32

[0243] This example provides a heterocyclic compound I-32, the reaction equation and preparation method of which are as follows:

[0244] Following a similar process to Example 31, 32.0 mg of the title compound was prepared from intermediate I-30 and starting material (S)-1-phenylethylamine as a white solid, with a yield of 50.4%.

[0245] The NMR and mass spectrometry data of the product are as follows:

[0246] 1H NMR(500MHz,DMSO-d6)δ8.58–8.44(m,2H),8.35(s,1H),8.03(d,J=8.3Hz,2H ),7.92(d,J=8.3Hz,2H),7.69(d,J=8.4Hz,1H),7.53–7.39(m,4H),7.37–7.21 (m,5H),7.18–7.04(m,2H),6.96–6.90(m,1H),5.21–5.01(m,1H),3.72–3.41( m,5H),3.30–3.03(m,3H),1.47(d,J=7.0Hz,3H).LC-MS(ESI):m / z640.2[M+H] + .

[0247] Example 33

[0248] This example provides a heterocyclic compound I-33, the reaction equation and preparation method of which are as follows:

[0249] Following a similar process to Example 31, 9.7 mg of the title compound was prepared from intermediate I-30 and starting material 1-thiomorpholine oxide as a white solid, with a yield of 15.1%.

[0250] The NMR and mass spectrometry data of the product are as follows:

[0251] 1 H NMR (500MHz, CDCl3) δ7.87–7.80(m,2H),7.67–7.56(m,3H),7.45(s,1H),7.37–7.31(m,3H),7.30–7.26(m,2H),7.16(d,J=7.2Hz ,1H),7.04(t,J=7.2Hz,1H),6.49(s,1H),4.33–3.90(m,4H),3.81–3.27(m,8H),2.99–2.56(m,2H),1.68(s,2H).LC-MS(ESI):m / z 638.2[M+H] + .

[0252] Example 34

[0253] This example provides a heterocyclic compound I-34, the reaction equation and preparation method of which are as follows:

[0254] Following a similar process to Example 31, 7.5 mg of the title compound was prepared from intermediate I-30 and starting material 1-thiomorpholine oxide as a white solid, with a yield of 11.4%.

[0255] The NMR and mass spectrometry data of the product are as follows:

[0256] 1 H NMR(500MHz, CDCl3)δ7.85(d,J=8.3Hz,2H),7.66–7.58(m,3H),7.46(s,1H),7.38–7.32(m,3H),7.31–7.26(m,2H),7.20– 7.16(m,1H),7.07–7.01(m,1H),6.50(s,1H),4.75–3.62(m,10H),3.45–2.59(m,4H),1.81–1.53(m,2H).LC-MS(ESI):m / z 654.2[M+H] + .

[0257] Example 35

[0258] This example provides a heterocyclic compound I-35, the reaction equation and preparation method of which are as follows:

[0259] Step 1: Intermediate I-1 (49.3 mg, 0.10 mmol, 1.0 equivalent) and starting material N-bromosuccinimide (17.8 mg, 0.10 mmol, 1.0 equivalent) were dissolved in 2 mL of N,N-dimethylformamide solution. After stirring the reaction solution at room temperature for 6 hours, the reaction was monitored by HPLC-MS until it was complete. The reaction was quenched by adding 5 mL of aqueous solution and then extracted three times with 15 mL of ethyl acetate. The organic phases were combined and washed with saturated sodium chloride aqueous solution and water. After drying with anhydrous sodium sulfate, the solution was concentrated under vacuum and separated and purified by preparative liquid chromatography to obtain 34.4 mg of yellow solid compound I-35, with a yield of 65.5%.

[0260] The NMR and mass spectrometry data of the product are as follows:

[0261] 1 H NMR (500MHz, CDCl3) δ7.87–7.83(m,2H),7.64–7.58(m,3H),7.47(s,1H),7.38–7.33(m,3H),7.32–7.29(m,2H),7.18(d,J=7.3Hz,1H),7.09 –7.05(m,1H),6.54(s,1H),4.10–4.02(m,1H),3.99–3.91(m,1H),3.84–3.77(m,1H),3.66–3.58(m,2H),3.45–3.30(m,3H).LC-MS(ESI):m / z 572.1[M+H] + .

[0262] Example 36

[0263] This example provides a heterocyclic compound I-36, whose reaction equation and preparation method are as follows:

[0264] Step 1: Intermediate I-1 (49.3 mg, 0.10 mmol, 1.0 equivalent) and starting material N-bromosuccinimide (89.0 mg, 0.50 mmol, 5.0 equivalent) were dissolved in 2 mL of N,N-dimethylformamide solution. The reaction solution was stirred overnight at room temperature. After the reaction was monitored by HPLC-MS until complete, 5 mL of aqueous solution was added to quench the reaction. The mixture was then extracted three times with 15 mL of ethyl acetate. The organic phases were combined and washed with saturated sodium chloride aqueous solution and water. After drying with anhydrous sodium sulfate, the mixture was concentrated under vacuum. The mixture was then separated and purified by preparative liquid chromatography to obtain 12.7 mg of yellow solid compound I-36, with a yield of 19.5%.

[0265] The NMR and mass spectrometry data of the product are as follows:

[0266] 1 H NMR(500MHz, CDCl3)δ7.83(d,J=8.2Hz,2H),7.61(d,J=8.2Hz,2H),7.59–7.52(m,2H),7.44(s,1H),7.34–7.24(m,3H),7.23– 7.17(m,2H),7.15–7.07(m,1H),4.01–3.89(m,1H),3.86–3.73(m,2H),3.68–3.49(m,2H),3.38–3.20(m,3H).LC-MS(ESI):m / z 650.0[M+H] + .

[0267] Performance testing

[0268] (1) Protein thermal migration experiment

[0269] Differential scanning fluorescence analysis was used to detect the thermal stability of the protein under different conditions, and the effect of the heterocyclic compound prepared in the examples on the thermal stability of the three isoforms of TEAD1 / 2 / 4 was investigated. A 20 μL mixture of 2.5 μM TEAD protein, 5×SYPRO Orange dye (Invitrogen), and 2.5 μM of the heterocyclic compound was added to a 96-well plate (DN Biotech). The wells were sealed with a heat-sealing film (Thermo Scientific) with good light transmittance. Subsequently, the thermal stability of the protein was assessed using QuantStudio. TMOn a 6Flex real-time PCR system (Applied Biosystems), following standard procedures, the reaction mixture was linearly heated from 25°C to 95°C within 25 minutes, with real-time monitoring of fluorescence signal intensity. Then, using Protein Themal Shift... TM Software Version 1.2 Life (Technologies) was used to fit the temperature-fluorescence intensity curve, i.e., the protein melting curve, and to determine the protein's melting temperature (Tm). In this experiment, DMSO was used as a negative control, and the change (ΔTm) of the compound on the Tm value of TEAD protein was calculated. The results are shown in Table 1. The compounds in the embodiments of this invention can significantly improve the thermal stability of TEAD1 / 2 / 4 proteins.

[0270] Table 1. Effects of heterocyclic compounds prepared in Examples 1-16 of this invention on the thermal stability of TEAD protein.

[0271] (2) Cancer cell growth inhibition experiment

[0272] NCI-H226 and NCI-H2452 cells were cultured in RPMI 1640 complete medium. Normally growing cells were digested with trypsin-based cell digestion solution, centrifuged, counted, and seeded into 96-well plates at a density of 1000 cells per well. After 24 hours of cell seeding, the cells were treated with 10 μL of an inhibitor at different concentration gradients per well, with three replicates for each concentration. The starting concentration was 20 μM, and the cells were diluted 4-fold each time. A corresponding 0.1% DMSO negative control group was also included. After 72 hours of treatment, the cell culture plates were removed from the incubator, the culture medium in the 96-well plates was aspirated, and 190 μL of RPMI 1640 complete medium was added again. The cells were then cultured again for 72 hours. The 96-well plates were then removed and allowed to equilibrate at room temperature for 10 minutes. The culture medium in the 96-well plates was aspirated, and 200 μL of CellTiter-Lumi was added. TM Chemiluminescence cell viability assay solution (mixed 1:1 with culture medium) was shaken for two minutes and then reacted at room temperature for 10 minutes. 150 μL of the solution was transferred from a clear 96-well plate to a 96-well white plate for chemiluminescence readings. Cell viability was calculated by subtracting the background value from the reading in each well. Viability (%) = (Sample / Vehicle-1)*100. Sample represents the chemiluminescence of the drug-treated group, and Vehicle represents the absorbance of the DMSO control group. Using GraphPad Prism 7.0 software, an S-shaped dose-viability curve was plotted using a nonlinear regression model, and the IC50 was calculated. 50 or GI 50 Values. The test results are shown in Table 2.

[0273] Table 2. Cell viability results of NCI-H266 and NCI-H2452 cells after compound treatment.

[0274] (3) Time-resolved fluorescence resonance energy transfer (TR-FERT) activity assay

[0275] TR-FRET experiments were performed in experimental buffer (50 mM HEPES pH 7.5, 200 mM NaCl, and 0.1% Pluronic F-68 solution). The synthesis method of TR-FRET tracer WZJ10 is described in the reference (J Med Chem. 2023, 66(7), 4617-4632). In the experiment, 100 nM His-TEAD-YBD recombinant protein was added to a 384-well plate, pretreated with compound I-1 prepared in Example 1 of the TEAD inhibitor for 5 h, then 800 nM WZJ-10 was added, and finally MAb Anti-6HIS Tb cryptate Gold htf (PerkinElmer, #61HI2TLA) was added to a final concentration of 50 ng / mL. TR-FRET signals (490 / 520 nm) were collected using a PHERAstar FSX plate reader (BMG Labtech). 50 The inhibition rate at a single concentration was obtained by processing with Graphpad Prism 8.0 software. The test results are shown in Figures 1a-1c and Table 3. The heterocyclic compound prepared in Example 1 exhibits strong inhibitory activity against TEAD1 / 2 / 4 of the TEAD family, with an IC50 value of [missing value]. 50 The effective values ​​were 64 nM, 19 nM, and 23 nM, respectively. The inhibitory activity data of other compounds against TEAD1 protein are shown in Table 3.

[0276] Table 3. Inhibitory activity data of compounds against TEAD1 protein.

[0277] (4) Rat pharmacokinetic test of the compounds of the present invention

[0278] As shown in Tables 4 and 5, six rats were divided into two groups, receiving intravenous injection and gavage administration of compound I-1 prepared in Example 1, respectively. In the intravenous injection group, approximately 0.25 mL of blood was collected from the jugular vein at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration. In the gavage group, approximately 0.25 mL of blood was collected at the same times. The concentration of compound I-1 in rat plasma samples was determined by LC-MS / MS, and pharmacokinetic parameters were calculated using WinNolin software. The intravenous injection and gavage administration doses of compound I-1 prepared in Example 1 were 1 mg / kg and 10 mg / kg, respectively. The pharmacokinetic parameters after administration are shown in Tables 4 and 5. Following a single intravenous injection of 1 mg / kg of compound I-1 and gavage administration of 10 mg / kg of compound I-1, the main pharmacokinetic parameters after intravenous administration were: C max The concentration was 667 ng / mL, T max It is 0.0833h, T 1 / 2 The AUC is 6.35h. 0-T 2287 hr*ng / mL, AUC 0-∞ The concentrations were 2536 hr*ng / mL, Vz was 3052 mL / kg, Cl was 467 mL / hr / kg, and MRT was... 0-t For 4.87 hours, MRT 0-∞ The duration of action was 6.99 h. The main pharmacokinetic parameters for gavage administration were: C max The concentration was 1313 ng / mL, T max For 2.00h, T 1 / 2 The AUC is 5.54h. 0-t The concentration was 12676 hr*ng / mL, with an AUC of 0-∞ The concentration was 13616 hr*ng / mL, MRT 0-t For 6.00h, MRT 0-∞ The time to oral bioavailability was 7.46 h, and the oral bioavailability was 53.7%, indicating good pharmacokinetic properties.

[0279] Table 4. Pharmacokinetic parameters of compound I-1 in rats after intravenous injection (n=3)

[0280] Table 5. Pharmacokinetic parameters of compound I-1 administered by gavage in rats (n=3)

[0281] The present invention has been described in detail above with reference to the embodiments of the present invention. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A heterocyclic compound, or a pharmaceutically acceptable salt thereof, its stereoisomers, its tautomers, its polymorphs, its solvates, its isotopic derivatives, or its prodrug, characterized in that, It has the structure shown in equation (Ⅰ): Where X is N, CH or -CBr; W is O or S; L is Or it may not exist; R1 is selected from phenyl, C 2～5 Ester-substituted phenyl, pyrimidinyl, cyclohexyl, naphthyl, and adamantyl groups; R2 is selected from H, halogen, aldehyde, carboxyl, and C. 1～10 alkyl, C 1～10 Halogenated alkyl, C 1～10 alkoxy groups, Among them, R5 and R6 are independently selected from H, Alternatively, R5 and R6 can form a saturated six-membered heterocycle doped with nitrogen and sulfur; V is selected from C or N; R7 and R8 are independently selected from H. Alternatively, R7 and R8 can form a saturated six-membered heterocycle doped with nitrogen and sulfur; R3 is selected from H, halogens, and C. 1～10 alkyl, C 1～10 Halogenated alkyl, C 1～10 alkoxy groups, R4 is selected from C 1～10 Alkyl, phenyl or C 1～10 Halogenated alkyl, C 1～10 Halogenated alkoxy groups, fluorinated sulfo-substituted phenyl groups, and C 1～10 Halogenated alkyl, C 1～10 Benzyl groups substituted with halogenated alkoxy groups.

2. The heterocyclic compound according to claim 1, or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, isotopic derivative, or prodrug, characterized in that, R2 is selected from H, aldehyde group, Among them, R5 and R6 are independently selected from H, Alternatively, R5 and R6 can form a saturated six-membered heterocycle doped with nitrogen and sulfur; V is selected from C or N; R7 and R8 are independently selected from H. Alternatively, R7 and R8 can form a saturated six-membered heterocycle doped with nitrogen and sulfur.

3. The heterocyclic compound according to claim 1, or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, isotopic derivative, or prodrug, characterized in that, R3 is selected from H, halogens, 4. The heterocyclic compound according to claim 1, or its pharmaceutically acceptable salt, stereoisomer, tautomer, polymorph, solvate, isotopic derivative, or prodrug, characterized in that, Selected from the following structural formulas:

5. A method for preparing the heterocyclic compound according to any one of claims 1 to 4, characterized in that, Includes the following steps: S1. Compound B is mixed with an acidic reagent and deprotected to obtain an intermediate; S2. The intermediate, compound A and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate are mixed and reacted to obtain the product. Alternatively, compound A and compound C react to obtain the product. Alternatively, compound D and compound E can be reacted to obtain the product. The structural formulas of compounds A, B, C, D, and E are as follows:

6. The preparation method according to claim 5, characterized in that, The acidic reagents include trifluoroacetic acid and / or hydrochloric acid.

7. A pharmaceutical composition, characterized in that, Includes the heterocyclic compound as described in any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, its stereoisomer, its tautomer, its polymorph, its solvate, its isotopic derivative, or its prodrug; And pharmaceutically acceptable excipients.

8. The pharmaceutical composition according to claim 7, characterized in that, The excipients include at least one of solvents, excipients, diluents, binders, disintegrants, dispersants, flavoring agents, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, preservatives, solid binders, flow aids, or lubricants.

9. The use of the heterocyclic compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, its stereoisomer, its tautomer, its polymorph, its solvate, its isotopic derivative, or its prodrug; or the use of the pharmaceutical composition of any one of claims 7 to 8 in the preparation of a medicament for the prevention and / or treatment of TEAD-mediated diseases or conditions.

10. The application according to claim 9, characterized in that, The diseases or conditions mediated by TEAD are selected from: colon cancer, diffuse large B-cell lymphoma, follicular lymphoma, leukemia, multiple myeloma, mesothelioma, gastric cancer, malignant rhabdoid tumor, hepatocellular carcinoma, prostate cancer, breast cancer, bile duct and gallbladder cancer, bladder cancer; brain tumors, neuroblastoma, schwannoma, glioma, glioblastoma and astrocytoma; cervical cancer, melanoma, endometrial cancer, esophageal cancer, head and neck cancer, lung cancer, nasopharyngeal carcinoma, ovarian cancer, pancreatic cancer, renal cell carcinoma, rectal cancer, thyroid cancer, parathyroid tumors, uterine tumors and soft tissue sarcomas, cardiovascular diseases, neurodegenerative diseases, malaria, AIDS, gout, diabetes, renal failure, and chronic lung diseases.

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