TEAD inhibitor

Abstract

The invention discloses a TEAD inhibitor, which comprises a compound as shown in a general formula (I) or a general formula (II), and a stereoisomer or a salt thereof: the two compounds break through the limitation that the existing inhibitor only depends on a single non-covalent interaction through a hydrophobic effect, a hydrophilic effect and a multi-dimensional mechanism of covalent binding; the invention provides two brand new skeletons of carboxylic acid and acrylamide, enriches the structural types of the TEAD inhibitor, realizes more efficient and more specific inhibition of TEAD transcriptional activity through accurate design of different regions of a palmitoylated pocket, provides a candidate compound library with various structures for developing high-activity and high-selectivity antitumor drugs, and has a wide application prospect. The problems of insufficient activity, low selectivity and single structure of inhibitors in the prior art are effectively solved.

Description

Technical Field

[0001] This invention belongs to the field of anticancer drug technology and relates to a TEAD inhibitor. Background Technology

[0002] The Hippo signaling pathway is one of the core mechanisms regulating organ development, tissue homeostasis, and regeneration and repair in organisms. As a conserved inhibitory pathway, it maintains organ size and functional stability by precisely regulating the balance between cell proliferation, differentiation, and apoptosis. The core effector molecules of this pathway are Yes-associated protein (YAP) and transcriptional enhanced associated domain protein (TEAD), which achieve dynamic subcellular localization regulation through phosphorylation modification. When the Hippo pathway is activated, YAP is phosphorylated and retained in the cytoplasm, either by binding to 14-3-3 protein or by proteasome degradation. When the Hippo signaling pathway is inhibited, YAP phosphorylation levels decrease, meaning that unphosphorylated YAP enters the nucleus and binds to TEAD to form a complex, activating the transcription and expression of downstream target genes that promote cell proliferation and inhibit apoptosis. This regulatory network plays a crucial role in both developmental biology and cancer development.

[0003] Recent studies have found that excessive activation of YAP is closely related to the occurrence and development of various malignant tumors. In solid tumors such as breast cancer, fallopian tube cancer, germ cell cancer, and kidney cancer, YAP drives abnormal tumor cell proliferation, evasion of apoptosis, and promotes metastasis by continuously activating TEAD-dependent transcriptional programs. Clinical data show that high YAP expression is significantly associated with poor prognosis in patients with gastric cancer, colon cancer, and other cancers. Therefore, targeted intervention of YAP-TEAD interaction is considered a highly promising anti-cancer strategy. As the core transcriptional partner of YAP, the activity of TEAD directly determines the expression intensity of downstream oncogenes, making it a key target for drug development.

[0004] Although initial progress has been made in the development of TEAD inhibitors, several challenges remain: First, the variety of inhibitors is limited, with only a few compounds currently in preclinical research, and most are structurally modified based on flufenamic acid backbones, resulting in insufficient chemical diversity and difficulty in overcoming efficacy bottlenecks. Second, the activity and selectivity of inhibitors need improvement; existing inhibitors exhibit only micromolar inhibitory activity against TEAD and may have off-target effects, limiting their in vivo antitumor efficacy. Summary of the Invention

[0005] In view of the problems existing in the prior art, the present invention provides a TEAD inhibitor, thereby solving the technical problems that the existing TEAD-targeting inhibitors have a single structural type, poor inhibitory activity against TEAD, and low selectivity.

[0006] This invention is achieved through the following technical solution:

[0007] A TEAD inhibitor comprising a compound of general formula (I), its stereoisomer or a salt thereof, or a compound of general formula (II), its stereoisomer or a salt thereof:

[0008]

[0009] Wherein, the R1 group is a substituted alkyl or substituted acyl group; the R2 group is one of H, methyl, ethyl and benzyl; X1 is any one of H, Cl and Br, and X2 is H or Cl;

[0010]

[0011] Wherein, the R3 group is a substituted alkyl or substituted acyl group; the X3 is H, Cl, Br, Any one of them; X4 is H or Cl.

[0012] Preferably, the R1 group is a cycloalkyl-substituted alkyl group, an aromatic ring-substituted alkyl group, an alkyl-substituted acyl group, or an aromatic ring-substituted acyl group.

[0013] Preferably, the R1 base is: as well as Any one of them.

[0014] Preferably, the R1 base is as well as Any one of them.

[0015] Preferably, the R3 group is a cycloalkyl-substituted alkyl group, an aromatic ring-substituted alkyl group, a cycloalkyl-substituted acyl group, or an aromatic ring-substituted acyl group.

[0016] Preferably, the R3 base is as well as Any one of them.

[0017] A pharmaceutical composition comprising one of the above-described TEAD inhibitors and one or more pharmaceutically acceptable carriers, diluents, or excipients; the pharmaceutical composition is formulated into tablets, capsules, soft capsules, or injections by adding excipients to one of the above-described TEAD inhibitors; the excipients include one or more of additives, stabilizers, solubilizers, lubricants, and disintegrants.

[0018] The above-mentioned TEAD inhibitor is used in the preparation of TEAD inhibitor drugs or drugs that inhibit TEAD-YAP protein-protein interactions.

[0019] The use of the above-mentioned TEAD inhibitor or the above-mentioned pharmaceutical composition in the preparation of cancer drugs; characterized in that the cancer includes any one of breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, gastric cancer, colorectal cancer, kidney cancer, ovarian cancer, uterine cancer, prostate cancer, glioma, myelodysplastic syndrome, and acute myeloid leukemia.

[0020] Compared with the prior art, the present invention has the following beneficial technical effects:

[0021] Based on the three-dimensional structural features of the TEAD palmitoylation pocket, this scheme designs novel derivatives comprising compounds of general formula (I) or general formula (II), their stereoisomers, or salts thereof: Among them, general formula (I) is a 2-phenoxyacetic acid derivative, and general formula (II) is an N-phenylacrylamide derivative. These two types of compounds achieve highly efficient and specific inhibition of TEAD through differentiated action modes. In the 2-phenoxyacetic acid derivative, the substituted phenyl moiety penetrates deep into the hydrophobic region inside the palmitoylation pocket. By introducing different substituents and key amino acid residues in the hydrophobic region (such as leucine and isoleucine), multiple hydrophobic interactions are formed, significantly enhancing the binding affinity. Simultaneously, its carboxylic acid group is precisely positioned in the hydrophilic region at the pocket entrance, forming hydrogen bonds or ionic interactions with conserved cysteine ​​and lysine residues, further stabilizing the complex structure. For the N-phenylacrylamide derivative, its acrylamide group covalently binds to the cysteine ​​residues at the palmitoylation site of TEAD through a Michael addition reaction, forming an irreversible inhibitory effect. This covalent binding mode not only significantly improves the inhibitory activity but also reduces the off-target risk by specifically targeting cysteine. These two types of compounds, through a multi-dimensional mechanism involving hydrophobic, hydrophilic, and covalent interactions, overcome the limitations of existing inhibitors that rely solely on single non-covalent interactions. They provide two novel skeletons—carboxylic acids and acrylamides—enriching the structural types of TEAD inhibitors. Furthermore, through precise design targeting different regions of the palmitoylation pocket (the hydrophobic core region and the hydrophilic entrance region), they achieve more efficient and specific inhibition of TEAD transcriptional activity. This provides a diverse library of candidate compounds for developing highly active and selective antitumor drugs, effectively solving the problems of insufficient inhibitor activity, low selectivity, and structural homogeneity in existing technologies. Detailed Implementation

[0022] To enable those skilled in the art to understand the features and effects of the present invention, the terms and expressions used in the specification and claims are explained and defined in general below. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in case of conflict, the definitions in this specification shall prevail.

[0023] The theories or mechanisms described and disclosed herein, whether right or wrong, should not in any way limit the scope of the invention, that is, the contents of the invention can be implemented without being limited by any particular theory or mechanism.

[0024] In this document, all features defined by numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are for the sake of brevity and convenience only. Accordingly, descriptions of numerical ranges or percentage ranges should be considered as covering and specifically disclosing all possible sub-ranges and individual numerical values ​​(including integers and fractions) within those ranges.

[0025] In this article, unless otherwise specified, “contains,” “includes,” “containing,” “has,” or similar terms cover the meanings of “composed of” and “mainly composed of,” for example, “A contains a” covers the meanings of “A contains a and others” and “A contains only a.”

[0026] For the sake of brevity, not all possible combinations of the technical features in each implementation scheme or embodiment are described herein. Therefore, as long as there is no contradiction in the combination of these technical features, the technical features in each implementation scheme or embodiment can be combined arbitrarily, and all possible combinations should be considered within the scope of this specification.

[0027] This invention provides a TEAD inhibitor, comprising a compound of general formula (I), its stereoisomer or a salt thereof, or a compound of general formula (II), its stereoisomer or a salt thereof:

[0028] General formula (I) is a 2-phenoxyacetic acid derivative, namely:

[0029]

[0030] Wherein, R1 is a substituted alkyl or substituted acyl group, R2 is any one of hydrogen, methyl, ethyl and benzyl, X1 is any one of hydrogen, chlorine and bromine, and X2 is hydrogen or chlorine.

[0031] Preferably, the R1 group is a cycloalkyl-substituted alkyl group, an aromatic ring-substituted alkyl group, an alkyl-substituted acyl group, or an aromatic ring-substituted acyl group.

[0032] Furthermore, the R1 base is Any one of them.

[0033] The R2 base is:

[0034] H, Any one of them.

[0035] General formula (II) is an N-phenylacrylamide derivative, namely:

[0036]

[0037] Wherein, the R3 group is a substituted alkyl group or a substituted acyl group;

[0038] X3 is H, Cl, Br. Any one of them.

[0039] X4 is H or Cl.

[0040] Preferably, the R3 group is a substituted alkyl group or a substituted acyl group;

[0041] More preferably, the R3 group is a cycloalkyl-substituted alkyl group, an aromatic ring-substituted alkyl group, a cycloalkyl-substituted acyl group, or an aromatic ring-substituted acyl group.

[0042] Furthermore, the R3 base is Any one of them.

[0043] In addition, the present invention discloses a pharmaceutical composition comprising a compound of general formula (I) or general formula (II) above, a stereoisomer of general formula (I) or general formula (II) or a salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients; the pharmaceutical composition is formulated into tablets, capsules, soft capsules or injections by adding excipients to the compound of general formula (I) or general formula (II) above, its stereoisomer or a salt thereof; the excipients include one or more of additives, stabilizers, solubilizers, lubricants and disintegrants. Each tablet, capsule or vial contains 10 to 500 mg of the compound of general formula (I) or (II) of the present invention, its stereoisomer or a salt thereof;

[0044] The present invention also discloses the use of the compounds represented by the above general formula (I) or (II), their stereoisomers or salts thereof, or the above pharmaceutical compositions in the preparation of TEAD inhibitor drugs and drugs that inhibit TEAD-YAP protein-protein interactions.

[0045] In addition, the present invention also discloses the use of the compounds represented by general formula (I) or (II), their stereoisomers or salts thereof, or the above-described pharmaceutical compositions in the preparation of cancer drugs; the cancers include any one of breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, gastric cancer, colorectal cancer, kidney cancer, ovarian cancer, uterine cancer, prostate cancer, glioma, myelodysplastic syndrome, and acute myeloid leukemia.

[0046] Furthermore, this invention also discloses a method for preparing the compound represented by the above general formula (I), its stereoisomers, or salts thereof, wherein when the R1 group is a substituted acyl group, the method includes the following steps:

[0047] S1: 2,3-Dichlorophenol is reacted with dimethyl sulfate to prepare 2,3-dichloro-anisole;

[0048] S2: React anisole, 2-chloroanisole, 3-chloroanisole, 2-bromoanisole, or 2,3-dichloroanisole with any one of different acyl chlorides to prepare anisole with hydrogen, chlorine, or bromine substitution at the 2-position, hydrogen or chlorine substitution at the 3-position, or different acyl groups at the 4-position.

[0049] S3: React the various anisoles obtained in step S2 with aluminum trichloride to prepare phenols with hydrogen, chlorine or bromine substitution at the 2-position, hydrogen or chlorine substitution at the 3-position, and different substituted acyl groups at the 4-position.

[0050] S4: React any of the phenols obtained in step S3 with ethyl bromoacetate, methyl (R / S)-2-hydroxypropionate, methyl (R / S)-2-hydroxybutyrate, or methyl (R / S)-2-hydroxy-3-phenylpropionate. The resulting product is then hydrolyzed to obtain a compound of general formula (I), a stereoisomer of general formula (I), or a salt thereof, wherein R1 is any one of different substituted acyl groups, R2 is any one of hydrogen, methyl, ethyl, or benzyl, X1 is any one of hydrogen, chlorine, or bromine, and X2 is any one of hydrogen or chlorine.

[0051] When the R1 group is any one of different substituted alkyl groups, the R2 group is hydrogen, X1 is chlorine, and X2 is chlorine, the 2,3-dichloroanisole with different substituted alkyl groups at the 4-position obtained in step S2 is reacted with triethylsilane to obtain the 2,3-dichloroanisole with different substituted alkyl groups at the 4-position. Then, the obtained product is reacted with boron tribromide to obtain the 2,3-dichlorophenol with different substituted alkyl groups at the 4-position. The obtained product is then reacted with ethyl bromoacetate. Finally, the obtained product is hydrolyzed to obtain a compound of general formula (I), a stereoisomer of general formula (I) or a salt thereof, wherein the R1 group is any one of different substituted alkyl groups, the R2 group is hydrogen, X1 is chlorine, and X2 is chlorine.

[0052] The preparation method of the compound represented by the above general formula (II), its stereoisomer or salt thereof includes the following steps:

[0053] When the R3 group is a substituted acyl group, X3 is any one of hydrogen, chlorine, and bromine, and the X4 group is any one of hydrogen and chlorine, the following steps are included:

[0054] S5: React the various phenols obtained in step S3 of claim 8 with 2-bromoisobutyramide to prepare anilines with hydrogen, chlorine or bromine substitution at the 2-position, hydrogen or chlorine substitution at the 3-position, and different substituted acyl groups at the 4-position;

[0055] S6: React the various anilines obtained in step S5 with acryloyl chloride to obtain compounds of general formula (II), stereoisomers of general formula (II) or their salts, wherein R3 group is any one of different substituted acyl groups, X3 is any one of hydrogen, chlorine or bromine, and X4 is any one of hydrogen or chlorine.

[0056] When the R3 group is either cyclohexylmethyl or benzyl, X3 is chlorine, and X4 is hydrogen, the 2-chloroaniline with 4-cyclohexylformyl or benzoyl group obtained in step S5 is reacted with triethylsilane to obtain 2-chloroaniline with 4-cyclohexylformyl or benzoyl group. The resulting product is then reacted with acryloyl chloride to obtain a compound of general formula (II), a stereoisomer of general formula (II), or a salt thereof, wherein the R3 group is either cyclohexylmethyl or benzyl, X3 is chlorine, and X4 is hydrogen.

[0057] When R3 is cyclohexylformyl, X3 is any one of phenyl, pyrazolyl, or pyridyl, and X2 is hydrogen, the 2-bromoaniline with the 4-cyclohexylformyl group obtained in step S5 is reacted with any one of pinacol phenylboronic acid, 1H-pyrazol-3-boronic acid, 1-methylpyrazol-3-boronic acid, 1-methyl-1H-pyrazol-5-boronic acid, or 3-pyridylboronic acid. The resulting product is then reacted with acryloyl chloride to prepare a compound of general formula (II), a stereoisomer of general formula (II), or a salt thereof, wherein R3 is cyclohexylformyl, X3 is any one of phenyl, pyrazolyl, or pyridyl, and X4 is hydrogen.

[0058] The structures of the compounds in this invention were determined by nuclear magnetic resonance (NMR) and / or gas chromatography-mass spectrometry (GC-MS) or high-resolution mass spectrometry. NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer or an AVANCEIII-600 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6) and deuterated chloroform (CDCl3) as the solvent and tetramethylsilane (TMS) as the internal standard. GC-MS measurements were performed using a Shimadzu GCMS-QP2010 GC-MS system. High-resolution mass spectrometry measurements were performed using a Thermo Fisher QExactive Plus high-resolution mass spectrometer. Qingdao GF254 silica gel plates were used for thin-layer chromatography (TLC), with a diameter of 0.15–0.20 mm. The diameter of the purified products was 0.4–0.5 mm. Column chromatography generally used 200–300 mesh Yantai Huanghai silica gel as the support. The starting materials used in the embodiments of this invention are known and commercially available, or can be synthesized using methods known in the art. Unless otherwise specified, all reactions of this invention are carried out under a dry nitrogen atmosphere with continuous magnetic stirring, using a dry solvent, and the reaction temperature is expressed in degrees Celsius.

[0059] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0060] The following examples use instruments and equipment conventional in the art. Experimental methods in the following examples, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. All raw materials used in the following examples are conventional commercially available products with specifications conventional in the art. In this specification and the following examples, unless otherwise specified, "%" refers to weight percentage, "parts" refers to parts by weight, and "ratio" refers to weight proportion.

[0061] A 2-phenoxyacetic acid derivative comprising a compound of general formula (I), its stereoisomers, or salts thereof:

[0062]

[0063] In general formula (I), R1 group is any one of substituted alkyl and substituted acyl groups, R2 group is any one of hydrogen, methyl, ethyl and benzyl, X1 is any one of hydrogen, chlorine and bromine, and X2 is any one of hydrogen and chlorine.

[0064] An N-phenylacrylamide derivative comprising the compound shown in (II), its stereoisomers, or salts thereof:

[0065]

[0066] In general formula (II), R3 is any one of substituted alkyl and substituted acyl groups, X3 is any one of hydrogen, chlorine, bromine, phenyl, pyrazolyl, and pyridinyl, and X4 is any one of hydrogen and chlorine.

[0067] Table 1 shows the specific structures of the 2-phenoxyacetic acid derivatives and N-phenylacrylamide derivatives obtained in Examples 1-40, as well as the characterization results of the products. The corresponding Chinese names of the 2-phenoxyacetic acid derivatives and N-phenylacrylamide derivatives obtained in Examples 1-40 are shown in Table 2.

[0068] Table 1. Characterization test data of the products in Examples 1-45

[0069]

[0070]

[0071]

[0072]

[0073]

[0074] The preparation methods of the products in each embodiment in Table 1 are given below:

[0075] Example 1

[0076]

[0077] Step 1: Preparation of 1,2-dichloro-3-methoxybenzene (Intermediate 1-b)

[0078] Weigh 1.63 g (10.00 mmol) of 2,3-dichlorophenol and 1.50 g (10.87 mmol) of potassium carbonate into a 250 mL round-bottom flask, add 10 mL of acetone to dissolve, stir magnetically for 30 min, and then add dimethyl sulfate (1.40 g, 11.11 mmol) dropwise through a constant pressure dropping funnel. After heating to 50–60 °C and reacting for 3 h, filter to remove potassium carbonate, wash the filter cake with acetone, evaporate the acetone under reduced pressure, dissolve in ethyl acetate, wash twice with saturated sodium bicarbonate and saturated sodium chloride solutions, dry with anhydrous sodium sulfate, filter, and evaporate the solvent under reduced pressure to obtain a white solid (1.67 g, 94%), i.e., intermediate 1-b.

[0079] HRMS:[MH] - 174.1274. 1 H NMR (400MHz, CDCl3) δ7.16(t,J=8.2Hz,1H),7.07(dd,J=8.2,1.4Hz,1H),6.84(dd,J=8.3,1.4Hz,1H),3.91(s,3H).

[0080] Step 2: Preparation of 1-(2,3-dichloro-4-hydroxyphenyl)butyl-1-one (intermediate 1-c)

[0081] Weigh intermediate 1-b (0.88 g, 4.97 mmol) into a 250 mL round-bottom flask, dissolve it in dichloromethane (15 mL), cool to 0 °C in an ice bath, add aluminum trichloride (0.70 g, 5.22 mmol), stir for 30 min, then slowly add n-butyryl chloride (0.56 g, 5.22 mmol), react in an ice bath for 1 h, then react at room temperature for 2.5 h, remove the solvent by vacuum distillation, add dichloromethane, and repeat the process twice. Add dichloromethane (30 mL) and aluminum trichloride (0.70 g, 5.22 mmol). React at 40-50 °C for 3 h. Cool to room temperature, place the reaction flask in an ice bath, and slowly add ice water to quench the reaction. Separate the organic phase. Extract the aqueous phase three times with dichloromethane. Combine the organic phases, wash twice with saturated sodium chloride, dry with anhydrous sodium carbonate, filter, remove the solvent under reduced pressure, and separate by silica gel column chromatography to obtain a white solid (0.83 g, 72%), i.e., intermediate 1-c.

[0082] HRMS:[MH] - 233.0129. 1 H NMR (400MHz, DMSO-d6) δ11.28 (s, 1H), 7.52 (d, J = 8.6Hz, 1H), 7.01

[0083] (d,J=8.6Hz,1H),2.86(t,J=7.1Hz,2H),1.58(h,J=7.3Hz,2H),0.89(t,J=7.4Hz,3H).

[0084] Step 3: Preparation of 2-(4-Butyryl-2,3-dichlorophenoxy)acetic acid (Example 1)

[0085] Weigh intermediate 1-c (0.50 g, 2.15 mmol) into a reaction flask, dissolve it in acetone (10 mL), add potassium carbonate (0.33 g, 2.36 mmol) and potassium iodide (0.07 g, 0.43 mmol) sequentially, stir at room temperature for 1 h, add ethyl bromoacetate (0.39 g, 2.36 mmol) dropwise, heat to 50–60 °C and react for 5 h, remove acetone by evaporation, add ethyl acetate, wash the organic phase twice with water, remove the solvent by evaporation under reduced pressure, dissolve in ethanol (10 mL), add water (10 mL) and potassium hydroxide (0.24 g, 4.29 mmol), react at room temperature for 2 h, remove ethanol by evaporation under reduced pressure, slowly add concentrated hydrochloric acid until pH = 1, a large amount of precipitate precipitates, filter, dry to obtain the product as a white solid (0.40 g, 65%), which is the product of Example 1.

[0086] HRMS:[MH] - 289.0043. 1H NMR (400MHz, DMSO-d6) δ13.26 (s, 1H), 7.62 (d, J = 8.7Hz, 1H), 7.14

[0087] (d,J=8.8Hz,1H),4.95(s,2H),2.89(t,J=7.1Hz,2H),1.59(h,J=7.3Hz,2H),0.90(t,J=7.4Hz,3H).

[0088] Example 2

[0089]

[0090] Weigh the product of Example 1 (0.30 g, 1.03 mmol), potassium carbonate (0.28 g, 2.06 mmol), and paraformaldehyde (0.06 g, 2.06 mmol) and dissolve them in 50% ethanol aqueous solution (10 mL). React at 90 °C for 10 h. Remove the ethanol under reduced pressure. Slowly add concentrated hydrochloric acid until pH = 1. A large amount of precipitate is formed. Filter, dry, and separate by silica gel column chromatography to obtain a white solid (0.30 g, 96%), which is the product of Example 2.

[0091] HRMS:[MH] - 301.0036. 1 H NMR(400MHz,DMSO-d6)δ7.33(d,J=8.6Hz,1H),7.13(d,J=8.7Hz,1H),6.07(s ,1H),5.57(s,1H),4.92(s,2H),2.37(q,J=7.4Hz,2H),1.08(t,J=7.4Hz,3H).

[0092] Example 3

[0093]

[0094] Weigh the product of Example 2 (0.605 g, 0.82 mmol) into a reaction flask, dissolve it in methanol (3 mL), add palladium on carbon (0.02 g, 0.20 mmol), react in a hydrogen atmosphere for 24 h, filter to remove palladium on carbon, separate by silica gel column chromatography, and obtain the product as a white solid (0.13 g, 53%), which is the product of Example 3.

[0095] HRMS:[MH] - 303.0191. 1H NMR (600MHz, DMSO-d6) δ7.57(d,J=8.7Hz,1H),7.12(d,J=8.7Hz,1H),4.92(s,2H),3.21(h,J=6.7Hz ,1H),1.71-1.61(m,1H),1.36(dp,J=14.4,7.3Hz,1H),1.06(d,J=6.9Hz,3H),0.85(t,J=7.4Hz,3H).

[0096] Example 4

[0097]

[0098] Step 1: Preparation of (2,3-dichloro-4-hydroxyphenyl)(phenyl)methyl ketone (intermediate 4-c)

[0099] The operation steps are the same as the second step in the preparation of Example 1, except that n-butyryl chloride is replaced with The reaction yielded a white solid (0.93 g, 70%), namely intermediate 4-c.

[0100] HRMS:[MH] - 266.9973. 1 H NMR (600MHz, CDCl3) δ7.80(d,J=6.9Hz,2H),7.61(t,J=7.4Hz,1H),7.47(t,J=7.8Hz,2H),7.26(d,J=8.4Hz,1H),7.05(d,J=8.5Hz,1H),6.07(s,1H).

[0101] Step 2: Preparation of 2-(4-benzoyl-2,3-dichlorophenoxy)acetic acid (Example 4)

[0102] The operation steps are the same as the third step in the preparation of Example 1, and the product obtained is a white solid (0.26 g, 44%), which is the product of Example 4.

[0103] HRMS:[MH] - 322.9885. 1 H NMR (400MHz, DMSO-d6) δ13.24(s,1H),7.77-7.66(m,3H),7.56(t,J=7.9Hz,2H),7.45(d,J=8.5Hz,1H),7.20(d,J=8.7Hz,1H),4.97(s,2H).

[0104] Example 5

[0105]

[0106] Step 1: Preparation of cyclohexyl(2,3-dichloro-4-hydroxyphenyl) methyl ketone (intermediate 5-c)

[0107] The operation steps are the same as the second step in the preparation of Example 1, except that n-butyryl chloride is replaced with The reaction yielded a white solid (1.15 g, 85%), namely intermediate 5-c.

[0108] HRMS:[MH] - 273.0441. 1 H NMR (400MHz, DMSO-d6) δ11.22 (s, 1H), 7.41 (d, J = 8.4Hz, 1H), 7.00

[0109] (d,J=8.5Hz,1H),3.03(q,J=6.2,4.1Hz,1H),1.83-1.54(m,5H),1.32-1.10(m,5H).

[0110] Step 2: Preparation of 2-(2,3-dichloro-4-(cyclohexylcarbonyl)phenoxy)acetic acid (Example 5)

[0111] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.09 g, 14%).

[0112] HRMS:[MH] - 329.0350. 1 H NMR(400MHz,DMSO-d6)δ7.53(d,J=8.7Hz,1H),7.12(d,J=8.7Hz,1H),4.94(s, 2H),3.05(ddq,J=10.6,7.0,3.5Hz,1H),1.82-1.57(m,5H),1.35-1.14(m,5H).

[0113] Example 6

[0114]

[0115] Step 1: Preparation of 1-(2,3-dichloro-4-hydroxyphenyl)-2-phenylethyl-1-one (intermediate 6-c)

[0116] The operation steps are the same as the second step in the preparation of Example 1, except that the corresponding reactants are replaced, and the product is a white solid (0.82 g, 59%).

[0117] HRMS:[MH] - 273.0441. 1H NMR (400MHz, DMSO-d6) δ11.40–11.29(m,1H),7.66(d,J=8.6Hz,1H),7.30(t,J=7.5Hz,2H),7.25-7.18(m,3H),7.01(d,J=8.6Hz,1H),4.25(s,2H).

[0118] Step 2: Preparation of 2-(2,3-dichloro-4-(2-phenylacetyl)phenoxy)acetic acid (Example 6)

[0119] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.19 g, 31%).

[0120] HRMS:[MH] - 337.0038. 1 H NMR (400MHz, DMSO-d6) δ7.77(d,J=8.8Hz,1H),7.31(ddd,J=7.6,6.2,1.6Hz,2H ),7.23(tt,J=8.0,1.5Hz,3H),7.15(d,J=8.8Hz,1H),4.95(s,2H),4.29(s,2H).

[0121] Example 7

[0122]

[0123] Step 1: Preparation of (E)-1-(2,3-dichloro-4-hydroxyphenyl)-3-phenylpropyl-2-en-1-one (Intermediate 7-c)

[0124] The operation steps are the same as the second step in the preparation of Example 1, except that the corresponding reactants are replaced, and the product is a white solid (0.87 g, 60%).

[0125] HRMS:[MH] - 293.0125. 1 H NMR (400MHz, DMSO-d6) δ11.30 (s, 1H), 7.80-7.74 (m, 2H), 7.50-7.41 (m, 5H), 7.30 (d, J = 16.1Hz, 1H), 7.06 (d, J = 8.5Hz, 1H).

[0126] Step 2: Preparation of 2-(2,3-dichloro-4-cinnamoylphenoxy)acetic acid (Example 7)

[0127] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.14 g, 24%).

[0128] HRMS:[MH] - 349.0036. 1 H NMR (400MHz, DMSO-d6) δ7.78 (dd, J=7.6, 2.2Hz, 2H), 7.57 (d, J=8.6

[0129] Hz,1H),7.52-7.39(m,4H),7.31(d,J=16.0Hz,1H),7.19(d,J=8.8Hz,1H),4.97(s,2H).

[0130] Example 8

[0131]

[0132] Step 1: Preparation of 1-(2,3-dichloro-4-hydroxyphenyl)-3-phenylpropyl-1-one (Intermediate 8-d)

[0133] The preparation procedure was the same as in Example 3, and the product was a white solid (0.36 g, 48%).

[0134] HRMS:[MH] - 295.0274. 1 H NMR(400MHz, DMSO-d6)δ11.32(s,1H),7.56(dd,J=8.6,2.3Hz,1H),7.31-7.20(m,4H),7.17(td ,J=6.9,2.0Hz,1H),7.00(dd,J=8.6,2.0Hz,1H),3.23(t,J=7.6Hz,2H),2.90(t,J=7.5Hz,2H).

[0135] Step 2: 2-(2,3-dichloro-4-(3-phenylpropionyl)phenoxy)acetic acid (Example 8)

[0136] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.40 g, 66%).

[0137] HRMS:[MH] - 351.0195. 1 H NMR (400MHz, CDCl3) δ7.35-7.26 (m, 3H), 7.21 (d, J = 7.3Hz, 3H), 6.77

[0138] (dd,J=8.6,1.1Hz,1H),4.80(s,2H),3.25(t,J=7.6Hz,2H),3.04(t,J=7.6Hz,2H).

[0139] Example 9

[0140]

[0141] Step 1: Preparation of (2,3-dichloro-4-methoxyphenyl)(phenyl)methyl ketone (intermediate 9-c)

[0142] Weigh intermediate 1-b (1.00 g, 5.65 mmol) into a reaction flask, dissolve it in dichloromethane (20 mL), cool to 0 °C in an ice bath, add aluminum trichloride (0.79 g, 5.92 mmol), stir for 30 min, then slowly add benzoyl chloride (0.83 g, 5.93 mmol), react in an ice bath for 1 h, then raise to room temperature and react for 2.5 h, cool to room temperature, place the reaction flask in an ice bath, slowly add ice water to the reaction flask, separate the organic phase, extract the aqueous phase three times with dichloromethane, combine the organic phases, wash twice with saturated sodium chloride, dry with anhydrous magnesium sulfate, filter, remove solvent by vacuum distillation, separate by silica gel column chromatography, and obtain the product as a white solid (1.31 g, 83%).

[0143] HRMS:[MH] - 281.0125. 1 H NMR (400MHz, CDCl3) δ7.84-7.74(m,2H),7.60(t,J=7.4Hz,1H),7.49-7.43(m,2H),7.30(d,J=8.5Hz,1H),6.94(d,J=8.5Hz,1H),3.99(s,3H).

[0144] Step 2: Preparation of 1-benzyl-2,3-dichloro-4-methoxybenzene (Intermediate 9-d)

[0145] Weigh intermediate 9-c (1.00 g, 3.56 mmol) into a reaction flask, dissolve it in a mixed solution of dichloromethane (10 mL) and trifluoroacetic acid (10 mL), add triethylsilane (1.03 g, 8.90 mmol), and stir the mixture at 50 °C for 12 h. After cooling the reaction mixture to room temperature, remove the solvent by evaporation and separate by silica gel column chromatography to obtain a white solid (0.76 g, 80%).

[0146] HRMS:[MH] - 267.2017. 1 H NMR (400MHz, DMSO-d6) δ7.33-7.25(m,3H),7.23–7.08(m,4H),4.05(s,2H),3.86(s,3H).

[0147] Step 3: Preparation of 4-benzyl-2,3-dichlorophenol (intermediate 9-e)

[0148] Intermediate 9-d (0.50 g, 1.87 mmol) was weighed into a reaction flask, dissolved in dichloromethane (10 mL), and cooled to 0 °C in an ice bath. BBr3 (0.70 g, 2.81 mmol) was added, and the reaction was continued in an ice bath for 30 min. The mixture was then quenched with cold water, extracted three times with ethyl acetate, and the combined organic layers were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. After evaporation to remove the solvent, the mixture was purified by silica gel column chromatography to give a white solid (0.42 g, 88%).

[0149] HRMS:[MH] - 259.0578. 1 H NMR (400MHz, DMSO-d6) δ10.50 (s, 1H), 7.34-7.23 (m, 2H), 7.22-7.09 (m, 4H), 6.92 (d, J = 8.5Hz, 1H), 4.00 (s, 2H).

[0150] Step 4: Preparation of 2-(4-benzyl-2,3-dichlorophenoxy)acetic acid (Example 9)

[0151] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.33g, 54%).

[0152] HRMS:[MH] - 309.0085. 1 H NMR (400MHz, DMSO-d6) δ7.30-7.24(m,2H),7.21-7.13(m,4H),6.81(d,J=8.6Hz,1H),4.29(s,2H),4.02(s,2H).

[0153] Example 10

[0154]

[0155] Step 1: Preparation of cyclohexyl(2,3-dichloro-4-methoxyphenyl) ketone (intermediate 10-c)

[0156] The procedure was performed as described in the first step of the preparation in Example 9, and the product was a white solid (1.22 g, 77%).

[0157] HRMS:[MH] - 287.0597. 1H NMR (400MHz, CDCl3) δ7.26 (d, J = 8.6 Hz, 1H), 6.87 (d, J = 8.6 Hz, 1H), 3.94 (s, 3H), 3.09-3.02 (m, 1H), 1.92-1.65 (m, 5H), 1.46-1.21 (m, 5H).

[0158] Step 2: Preparation of 2,3-dichloro-1-(cyclohexylmethyl)-4-methoxybenzene (intermediate 10-d)

[0159] The operation steps are the same as the second step in the preparation of Example 9, and the product is a white solid (0.73 g, 77%).

[0160] HRMS:[MH] - 273.1675. 1 H NMR (400MHz, DMSO-d6) δ7.22(d,J=8.5Hz,1H),7.08(d,J=8.6Hz,1H),3.85(s,3H),2.57(d,J=6.9Hz,2H),1.72-1.45(m,6H),1.17-0.90(m,5H).

[0161] Step 3: Preparation of 2,3-dichloro-4-(cyclohexylmethyl)phenol (intermediate 10-e)

[0162] The operation steps are the same as the third step in the preparation of Example 9, and the product is a white solid (0.37 g, 79%).

[0163] HRMS:[MH] - 253.0528. 1 H NMR (400MHz, DMSO-d6) δ10.37 (s, 1H), 7.03 (d, J = 8.4Hz, 1H), 6.87

[0164] (d,J=8.4Hz,1H),2.51(d,J=6.9Hz,2H),1.74-1.54(m,5H),1.50(ddd,J=11.0,7.3,3.6Hz,1H),1.23-1.06(m,3H),0.94(tt,J=14.9,6.9Hz,2H).

[0165] Step 4: Preparation of 2-(2,3-dichloro-4-(cyclohexylmethyl)phenoxy)acetic acid (Example 10)

[0166] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.42 g, 70%).

[0167] HRMS:[MH] - 315.0559. 1 H NMR (400MHz, DMSO-d6) δ7.18(d,J=8.6Hz,1H),6.96(d,J=8.7Hz,1H),4.82(s,2H),2. 56(d,J=6.8Hz,2H),1.68-1.47(m,6H),1.12(q,J=8.8,8.1Hz,3H),1.02-0.88(m,2H).

[0168] Example 11

[0169]

[0170] Step 1: Preparation of 1-(2,3-dichloro-4-methoxyphenyl)-2-phenethyl-1-one (intermediate 11-c)

[0171] The procedure was performed as described in the first step of the preparation in Example 9, and the product was a white solid (0.96 g, 58%).

[0172] HRMS:[MH] - 293.0286. 1 H NMR (400MHz, CDCl3) δ7.35 (d, J=8.7Hz, 1H), 7.30 (dd, J=7.3, 5.3Hz, 2H), 7.27-7. 25(m,1H),7.22(d,J=7.1Hz,2H),6.84(d,J=8.6Hz,1H),4.25(s,2H),3.94(s,3H).

[0173] Step 2: Preparation of 2,3-dichloro-1-methoxy-4-phenylethylbenzene (intermediate 11-d)

[0174] The operation steps are the same as the second step in the preparation of Example 9, and the product is a white solid (0.79 g, 83%).

[0175] HRMS:[MH] - 281.0511. 1 H NMR (400MHz, CDCl3) δ7.35-7.27(m,2H),7.24-7.15(m,3H),6.99(d,J=8.5Hz, 1H), 6.76 (d, J = 8.5Hz, 1H), 3.89 (s, 3H), 3.06-2.96 (m, 2H), 2.93-2.83 (m, 2H).

[0176] Step 3: Preparation of 2,3-dichloro-4-phenylethylphenol (intermediate 11-e)

[0177] The operation steps are the same as the third step in the preparation of Example 9, and the product is a white solid (0.38 g, 80%).

[0178] HRMS:[MH] - 267.0852. 1 H NMR(400MHz,DMSO-d6)δ10.43(s,1H),7.33-7.24(m,2H),7.24-7.14(m,3H),7.0 8(d,J=8.5Hz,1H),6.86(d,J=8.4Hz,1H),2.95-2.86(m,2H),2.85-2.75(m,2H).

[0179] Step 4: Preparation of 2-(2,3-dichloro-4-phenethylphenoxy)acetic acid (Example 11)

[0180] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.40 g, 66%).

[0181] HRMS:[MH] - 323.0249. 1 H NMR (400MHz, DMSO-d6) δ7.38-7.07(m,6H),6.74(d,J=8.6Hz,1H),4.20(s,2H),2.92(dd,J=10.2,5.6Hz,2H),2.81(dd,J=10.1,5.8Hz,2H).

[0182] Example 12

[0183]

[0184] Step 1: Preparation of 2,3-dichloro-4-(2-phenylcyclopropyl)phenol (Intermediate 12-d)

[0185] Intermediate 7-C (0.50 g, 1.71 mmol), potassium hydroxide (0.29 g, 5.12 mmol), and hydrazine hydrate (0.26 g, 0.25 mmol) were weighed and dissolved in diethylene glycol (10 mL). The mixture was refluxed for 1 h. The condenser was removed, and the reaction temperature was raised to 200 °C. The condenser was then reinserted, and the reaction was maintained at 200 °C for another 3 h. The reaction was monitored by TLC until complete. The mixture was cooled to room temperature, diluted with water (50 mL), and concentrated hydrochloric acid was added dropwise until the pH reached 1. The mixture was extracted three times with dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate. After evaporation of the solvent, the mixture was separated by silica gel column chromatography to obtain a yellow solid (0.16 g, 35%).

[0186] HRMS:[MH]- 279.0932. 1 H NMR (400MHz, DMSO-d6) δ7.34-7.11(m,6H),6.99(d,J=8.7Hz,1H),4.84(s,2H),2.32(dt,J=8 .8,5.8Hz,1H),2.08(dt,J=8.9,5.5Hz,1H),1.52(dt,J=8.8,5.6Hz,1H),1.47-1.38(m,1H).

[0187] Step 2: Preparation of 2-(2,3-dichloro-4-(2-phenylcyclopropyl)phenoxy)acetic acid (Example 12)

[0188] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.45 g, 74%).

[0189] HRMS:[MH] - 335.0250. 1 H NMR (400MHz, DMSO-d6) δ7.29(t,J=7.5Hz,2H),7.24-7.14(m,4H),6.99(d,J=8.7Hz,1H),4.84(s,2H),2.32( dt,J=8.8,5.8Hz,1H),2.08(dt,J=8.9,5.4Hz,1H),1.52(dt,J=8.8,5.6Hz,1H),1.44(dt,J=8.7,5.5Hz,1H).

[0190] Example 13

[0191]

[0192] Step 1: Preparation of 2,3-dichloro-4-(3-phenylpropyl)phenol (intermediate 13e)

[0193] The operation steps are the same as the third step in the preparation of Example 9, and the product obtained is a brown solid (0.13g, 27%).

[0194] HRMS:[MH] - 280.0420. 1 H NMR (400MHz, DMSO-d6) δ11.33 (s, 1H), 7.56 (d, J = 8.6Hz, 1H), 7.31-7.20 (m, 4H), 7.17 (t,J=7.0Hz,1H),6.99(d,J=8.6Hz,1H),3.23(t,J=7.5Hz,2H),2.90(t,J=7.5Hz,2H).

[0195] Step 2: Preparation of 2-(2,3-dichloro-4-(3-phenylpropyl)phenoxy)acetic acid (Example 13)

[0196] The operation steps are the same as the third step in the preparation of Example 1, and the product is a white solid (0.26 g, 44%).

[0197] HRMS:[MH] - 377.0379. 1 H NMR (400MHz, DMSO-d6) δ7.28(t,J=7.6Hz,2H),7.25(d,J=8.6Hz,1H),7.21(d,J=6.7Hz,2H),7.18(t,J=7.3H z,1H),6.98(d,J=8.7Hz,1H),4.82(s,2H),2.77-2.67(m,2H),2.63(t,J=7.7Hz,2H),1.84(p,J=7.8Hz,2H).

[0198] Example 14

[0199]

[0200] Weigh intermediate 5-C (0.20 g, 0.73 mmol), PPh3 (0.29 g, 1.11 mmol), and (D)-methyl lactate (0.11 g, 1.11 mmol) and dissolve them in THF (10 mL). Cool the mixture to 0 °C in an ice bath and slowly add DIAD (0.22 g, 1.11 mmol). Then, raise the mixture to room temperature and react for 1 h. Remove the solvent under reduced pressure and dissolve the mixture in ethanol (5 mL). Add water (5 mL) and potassium hydroxide (0.21 g, 3.74 mmol) and react at room temperature for 2 h. Remove the ethanol under reduced pressure and slowly add concentrated hydrochloric acid until pH = 1. A large amount of precipitate precipitates out. Filter and dry to obtain the product as a white solid (0.08 g, 33%).

[0201] HRMS:[MH] - 343.0507. 1 H NMR (400MHz, DMSO-d6) δ13.32 (s, 1H), 7.53 (d, J = 8.7Hz, 1H), 7.05

[0202] (d,J=8.7Hz,1H),5.12(q,J=6.8Hz,1H),3.05(td,J=10.0,8.5,4.9Hz,1H),1.83-1.62(m,5H),1.57(d,J=6.8Hz,3H),1.35-1.11(m,5H).

[0203] Example 15

[0204]

[0205] The preparation procedure was the same as in Example 14, and the product was a white solid (0.09 g, 36%).

[0206] HRMS:[MH] - 343.0510. 1 H NMR (400MHz, DMSO-d6) δ13.29 (s, 1H), 7.53 (d, J = 8.6Hz, 1H), 7.05

[0207] (d,J=8.8Hz,1H),5.12(q,J=6.8Hz,1H),3.05(ddd,J=11.0,7.3,3.9Hz,1H),1.82-1.62(m,5H),1.57(d,J=6.7Hz,3H),1.38-1.08(m,5H).

[0208] Example 16

[0209]

[0210] Step 1: Preparation of cyclopentyl(2,3-dichloro-4-hydroxyphenyl) ketone (intermediate 20-c)

[0211] The operation steps are the same as the second step in the preparation of Example 1, and the product obtained is a colorless oily liquid (1.31 g, 90%).

[0212] HRMS:[MH] - 258.0212. 1 H NMR(400MHz, DMSO-d6)δ7.59(d,J=8.8Hz,1H),6.90(d,J=8.8Hz,1H),1.98-1.79(m,6H),1.76-1.59(m,5H)

[0213] Step 2: Preparation of (R)-2-(2,3-dichloro-4-(cyclopentylcarbonyl)phenoxy)propionic acid (Example 16)

[0214] The preparation procedure was the same as in Example 14, and the product was a white solid (0.14 g, 22%).

[0215] HRMS:[MH] - 329.2078. 1 H NMR (600MHz, DMSO-d6) δ13.30 (s, 1H), 7.59 (d, J = 8.7Hz, 1H), 7.05

[0216] (d,J=8.8Hz,1H),5.12(q,J=7.2,6.6Hz,1H),3.59(dt,J=14.3,6.8Hz,1H),1.82-1.68(m,4H),1.58(d,J=8.7Hz,7H).

[0217] Example 17

[0218]

[0219] The preparation procedure was the same as in Example 14, and the product was a white solid (0.21 g, 32%).

[0220] HRMS:[MH] - 343.1451. 1 H NMR (600MHz, DMSO-d6) δ13.32 (s, 1H), 7.59 (d, J = 8.7Hz, 1H), 7.03

[0221] (d,J=8.7Hz,1H),4.98(t,J=5.8Hz,1H),3.59(p,J=7.8Hz,1H),1.97(ddq,J=28.7,14.2, 7.8,7.0Hz,2H),1.83-1.67(m,4H),1.58(dd,J=11.9,4.7Hz,4H),1.03(t,J=7.4Hz,3H).

[0222] Example 18

[0223]

[0224] Step 1: Preparation of (4-hydroxyphenyl)(phenyl)methyl ketone (intermediate 22-b)

[0225] The operation steps are the same as the second step in the preparation of Example 1, and the product is a slightly yellow transparent oily liquid (1.75 g, 93%).

[0226] HRMS:[MH] - 205.1216. 1 H NMR (400MHz, DMSO-d6) δ10.27 (s, 1H), 7.80 (d, J = 8.6Hz, 2H), 6.80

[0227] (d,J=8.7Hz,2H),3.24(dd,J=11.0,2.8Hz,1H),1.75-1.61(m,5H),1.42-1.15(m,5H).

[0228] Step 2: Preparation of (R)-2-(4-(cyclohexylcarbonyl)phenoxy)propionic acid (Example 18)

[0229] The preparation procedure was the same as in Example 14, and the product was a white solid (0.20 g, 30%).

[0230] HRMS:[MH] - 275.1285. 1 H NMR (400MHz, DMSO-d6) δ12.36 (s, 1H), 7.92 (d, J = 9.0Hz, 2H), 6.96

[0231] (d,J=8.8Hz,2H),4.98(q,J=6.8Hz,1H),3.33-3.28(m,1H),1.80-1.63(m,5H),1.53(d,J=6.8Hz,3H),1.46-1.12(m,5H).

[0232] Example 19

[0233]

[0234] Step 1: Preparation of (3-chloro-4-hydroxyphenyl)(cyclohexyl) methyl ketone (intermediate 23-b)

[0235] The operation steps are the same as the second step in the preparation of Example 1, and the product obtained is a colorless oily liquid (1.46 g, 88%).

[0236] HRMS:[MH] - 239.0832. 1 H NMR (400MHz, DMSO-d6) δ11.22 (s, 1H), 7.41 (d, J = 8.4Hz, 1H), 7.00

[0237] (d,J=8.5Hz,1H),3.03(q,J=6.2,4.1Hz,1H),1.83-1.54(m,5H),1.32-1.10(m,5H).

[0238] Step 2: Preparation of (R)-2-(2-chloro-4-(cyclohexylcarbonyl)phenoxy)propionic acid (Example 19)

[0239] The preparation procedure was the same as in Example 14, and the product was a white solid (0.18 g, 29%).

[0240] HRMS:[MH] - 309.0925. 1H NMR (400MHz, DMSO-d6) δ12.39 (s, 1H), 7.98 (d, J = 2.2Hz, 1H), 7.91

[0241] (dd,J=8.8,2.2Hz,1H),7.06(d,J=8.8Hz,1H),5.11(q,J=6.8Hz,1H),3.39 -3.35(m,1H),1.79-1.62(m,5H),1.57(d,J=6.7Hz,3H),1.49-1.11(m,5H).

[0242] Example 20

[0243]

[0244] Step 1: Preparation of (2-chloro-4-hydroxyphenyl)(cyclohexyl) methyl ketone (intermediate 24-b)

[0245] The operation steps are the same as the second step in the preparation of Example 1, and the product obtained is a colorless oily liquid (1.50 g, 90%).

[0246] HRMS:[MH] - 239.0831. 1 H NMR (600MHz, CDCl3) δ7.34(d,J=8.4Hz,1H),6.90(d,J=2.4Hz,1H),6.76(dd,J=8.5,2.4Hz,1H),3.14(tt,J=11.3,3.4Hz, 1H), 1.90 (dd, J=13.5, 3.2Hz, 2H), 1.80 (dt, J=12.8, 3.5Hz, 2H), 1.71-1.64 (m, 1H), 1.47-1.38 (m, 2H), 1.35-1.21 (m, 3H).

[0247] Step 2: Preparation of (R)-2-(3-chloro-4-(cyclohexylcarbonyl)phenoxy)propionic acid (Example 20)

[0248] The preparation procedure was the same as in Example 14, and the product was a white solid (0.26 g, 40%).

[0249] HRMS:[MH] - 309.0898. 1H NMR (600MHz, CDCl3) δ7.37(d,J=8.6Hz,1H),6.92(d,J=2.5Hz,1H),6.79(dd,J=8.6,2.5Hz,1H),4.81(q,J=6.8Hz,1H),3.10(tt ,J=11.3,3.4Hz,1H),1.92-1.85(m,2H),1.78(dt,J=12.2,3.5Hz,2H),1.70-1.61(m,4H),1.47-1.37(m,2H),1.34-1.20(m,3H).

[0250] Example 21

[0251]

[0252] Step 1: Preparation of (3-bromo-4-hydroxyphenyl)(cyclohexyl) methyl ketone (intermediate 25-b)

[0253] The operation steps are the same as the second step in the preparation of Example 1, and the product is a white solid (2.60 g, 87%).

[0254] HRMS:[MH] - 283.0328. 1 H NMR (400MHz, DMSO-d6) δ11.21 (s, 1H), 8.04 (d, J = 2.1Hz, 1H), 7.83

[0255] (dd,J=8.6,2.2Hz,1H),7.02(d,J=8.5Hz,1H),3.28(dt,J=11.1,3.0Hz,1H),1.82-1.63(m,5H),1.43-1.06(m,5H).

[0256] Step 2: Preparation of (R)-2-(2-bromo-4-(cyclohexylcarbonyl)phenoxy)propionic acid (Example 21)

[0257] The preparation procedure was the same as in Example 14, and the product was a white solid (0.20 g, 32%).

[0258] HRMS:[MH] - 353.0379. 1 H NMR (600MHz, DMSO-d6) δ13.28 (s, 1H), 8.12 (d, J = 2.2Hz, 1H), 7.95

[0259] (d,J=9.7Hz,1H),7.02(d,J=8.7Hz,1H),5.10(q,J=6.7Hz,1H),3.36(dt,J=11.2,2.8Hz,1H),1.80-1.69(m,4H ),1.66(d,J=13.5Hz,1H),1.58(d,J=6.7Hz,3H),1.41(q,J=12.2Hz,2H),1.35-1.27(m,2H),1.24-1.14(m,1H).

[0260] Example 22

[0261]

[0262] The preparation procedure was the same as in Example 14, and the product was a white solid (0.30 g, 42%).

[0263] HRMS:[MH] - 289.1438. 1 H NMR (600MHz, CDCl3) δ7.92(d,J=8.8Hz,2H),6.92(d,J=8.9Hz,2H),4.69(dd,J=7.0,5.1Hz,1H),3.20(tt,J=11.6,3.2Hz,1H),2.10-2.02(m,2H),1. 84(tt,J=13.7,2.6Hz,4H),1.73(dt,J=13.0,1.6Hz,1H),1.53-1.44(m,2H ),1.42-1.33(m,2H),1.27(tt,J=12.6,3.6Hz,1H),1.11(t,J=7.4Hz,3H).

[0264] Example 23

[0265]

[0266] The preparation procedure was the same as in Example 14, and the product was a white solid (0.19 g, 29%).

[0267] HRMS:[MH] - 323.1053. 1H NMR (600MHz, CDCl3) δ8.13(s,1H),7.37(d,J=8.6Hz,1H),6.93(d,J=2.5Hz,1H),6 .79(dd,J=8.6,2.5Hz,1H),4.63(dd,J=7.1,4.9Hz,1H),3.10(tt,J=11.3,3.4Hz, 1H),2.12-1.97(m,2H),1.93-1.84(m,2H),1.78(dt,J=12.7,3.4Hz,2H),1.71-1. 63(m,1H),1.41(qt,J=12.3,3.6Hz,2H),1.33-1.20(m,3H),1.10(t,J=7.4Hz,3H).

[0268] Example 24

[0269]

[0270] The preparation procedure was the same as in Example 14, and the product was a white solid (0.13 g, 20%).

[0271] HRMS:[MH] - 367.0545. 1 H NMR (600MHz, DMSO-d6) δ13.31(s,1H),8.12(d,J=2.2Hz,1H),7.99-7.93(m,1H),7.00(d,J=8.6Hz,1H),4.95(dd,J=6.9,4.6Hz,1H),3.39-3.33 (m,1H),2.04-1.91(m,2H),1.80-1.65(m,5H),1.41(q,J=12.6Hz,2H),1.34-1.27(m,2H),1.19(p,J=13.1,12.6Hz,1H),1.04(t,J=7.4Hz,3H).

[0272] Example 25

[0273]

[0274] Step 1: Preparation of cyclohexyl(4-amino-2,3-dichlorophenyl) ketone (intermediate 30-d)

[0275] Intermediate 5-C (1.00 g, 4.22 mmol), 2-bromoisobutyramide (2.09 g, 12.65 mmol), and sodium hydroxide (0.51 g, 12.65 mmol) were weighed and placed sequentially into a reaction flask. 21 mL of DMA was added, and the mixture was stirred at room temperature for 12 h. After the reaction was complete, sodium hydroxide (1.52 g, 37.95 mmol) was added, and the mixture was heated to 70 °C under N2 protection and reacted for 13 h. 8 mL of water was added, and the mixture was heated under reflux for 10 h. After cooling to room temperature, the mixture was extracted with ethyl acetate, and the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to obtain a white solid (0.21 g, 21%).

[0276] EI-MS: [M] + 271.05. 1 H NMR (400MHz, DMSO-d6) δ7.36(d,J=8.5Hz,1H),6.76(d,J=8.6Hz,1H),6.23(s,2H),3.08(qd,J=6.9,3.1Hz,1H),1.81-1.51(m,5H),1.41-1.07(m,5H).

[0277] Step 2: Preparation of N-(2,3-dichloro-4-(cyclohexylcarbonyl)phenyl)acrylamide (Example 25)

[0278] Weigh 30-d (0.35 g, 1.29 mmol) into a reaction flask, dissolve in dichloromethane (6 mL), cool to 0 °C in an ice bath, and slowly add triethylamine (0.14 g, 1.40 mmol) and acryloyl chloride (0.13 g, 1.40 mmol). React the mixture at room temperature for 1 h, extract with saturated sodium bicarbonate solution and dichloromethane, dry to anhydrous sodium sulfate, filter, and separate by silica gel column chromatography to obtain the product as a white solid (0.15 g, 36%).

[0279] EI-MS: [M] + 325.10. 1 H NMR (400MHz, DMSO-d6) δ7.61(d,J=8.3Hz,1H),7.50(d,J=8.4Hz,1H),6.60(dd,J=17.3,1.2Hz,1H),6.50-6. 44(m,1H),6.24(dd,J=10.2,1.2Hz,1H),3.01(tt,J=10.7,3.5Hz,1H),1.84-1.53(m,5H),1.35-1.09(m,5H).

[0280] Example 26

[0281]

[0282] Step 1: Preparation of cyclohexyl(4-aminophenyl) methyl ketone (intermediate 31-c)

[0283] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (1.59 g, 50%).

[0284] EI-MS: [M] + 203.15. 1 H NMR (400MHz, DMSO-d6) δ7.67(d,J=8.8Hz,2H),6.56(d,J=8.7Hz,2H),6.00(s,2H),3.22(d,J=14.5Hz,1H),1.75-1.65(m,5H),1.40-1.15(m,5H).

[0285] Step 2: Preparation of N-(4-cyclohexylcarbonyl)phenyl)acrylamide (Example 26)

[0286] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.42 g, 56%).

[0287] EI-MS: [M] + 257.15. 1 H NMR (400MHz, DMSO-d6) δ7.94(d,J=8.8Hz,2H),7.78(d,J=8.8Hz,2H),6.44(dd,J=17.0,10.1Hz,1H),6.28(d d,J=17.0,2.0Hz,1H),5.80(dd,J=10.0,2.0Hz,1H),3.29(s,1H),1.75(d,J=10.9Hz,5H),1.51-1.11(m,5H).

[0288] Example 27

[0289]

[0290] Step 1: Preparation of 4-benzyl-2-chloroaniline (intermediate 32-c)

[0291] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (0.41 g, 41%).

[0292] EI-MS: [M] + 237.10. 1H NMR (400MHz, DMSO-d6) δ7.77(d,J=2.0Hz,1H),7.67(dd,J=8.5,2.0Hz,1H),6.79(d,J= 8.5Hz,1H),6.26(s,2H),3.31-3.17(m,1H),1.68(t,J=13.4Hz,5H),1.43-1.09(m,5H).

[0293] Step 2: Preparation of N-(2-chloro-4-(cyclohexylcarbonyl)phenyl)acrylamide (Example 27)

[0294] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.11 g, 64%).

[0295] EI-MS: [M] + 291.10. 1 H NMR (400MHz, DMSO-d6) δ9.86(s,1H),8.11(d,J=8.5Hz,1H),8.02(d,J=2.0Hz,1H),7.94(dd,J=8.6,2.0Hz,1H),6.72(dd,J=17.0,10.2H z,1H),6.32(dd,J=17.0,1.9Hz,1H),5.84(dd,J=10.2,1.9Hz,1H),3.39(tt,J=11.1,3.2Hz,1H),1.91-1.64(m,5H),1.53-1.00(m,5H).

[0296] Example 28

[0297]

[0298] Step 1: Preparation of cyclopentyl(3-chloro-4-hydroxyphenyl) ketone (intermediate 33-b)

[0299] The operation steps are the same as the second step in the preparation of Example 1, and the product is a white solid (1.90 g, 80%).

[0300] EI-MS: [M] + 224.10. 1 H NMR (400MHz, DMSO-d6) δ11.15(s,1H),7.93(d,J=2.1Hz,1H),7.82(dd,J=8.5,2.2Hz,1H),7. 05(d,J=8.5Hz,1H),3.78-3.58(m,1H),1.85(tq,J=12.4,5.8,4.3Hz,2H),1.78-1.53(m,6H).

[0301] Step 2: Preparation of cyclopentanehexane (4-amino-3-chlorophenyl) ketone (intermediate 33-c)

[0302] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (0.80 g, 42%).

[0303] EI-MS: [M] + 223.10. 1 H NMR(400MHz,DMSO-d6)δ7.80(d,J=2.1Hz,1H),7.69(dd,J=8.5,2.1Hz,1H),6.80(d,J=8.6Hz ,1H),6.25(s,2H),3.66(ddd,J=15.7,8.6,6.8Hz,1H),1.88-1.79(m,2H),1.69-1.54(m,6H).

[0304] Step 3: Preparation of N-(2-chloro-4-(cyclopentylcarbonyl)phenyl)acrylamide (Example 28)

[0305] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.53 g, 54%).

[0306] EI-MS: [M] + 277.05. 1 H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 8.08 (d, J = 8.7Hz, 1H), 8.01 (s, 1H), 7.93 (d, J = 10.5Hz, 1H), 6.75-6.63 (m, 1H), 6.29 (dd, J = 17. 0,1.9Hz,1H),5.81(dd,J=10.3,1.8Hz,1H),3.85-3.73(m,1H),1.85(dq,J=12.7,6.4Hz,2H),1.75-1.62(m,2H),1.62-1.51(m,4H).

[0307] Example 29

[0308]

[0309] Step 1: Preparation of cyclopentyl(3-chloro-4-hydroxyphenyl) ketone (intermediate 34-b)

[0310] The operation steps are the same as the second step in the preparation of Example 1, and the product is a white solid (1.35 g, 83%).

[0311] EI-MS: [M]+ 232.05. 1 H NMR (400MHz, DMSO-d6) δ11.29 (s, 1H), 7.86-7.65 (m, 4H), 7.65-7.54 (m, 3H), 7.10 (d, J = 8.4Hz, 1H).

[0312] Step 2: Preparation of (4-amino-3-bromophenyl)(phenyl)methyl ketone (intermediate 34-c)

[0313] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (0.60 g, 44%).

[0314] EI-MS: [M] + 231.05. 1 H NMR (400MHz, DMSO-d6) δ7.72-7.57(m,4H),7.57-7.38(m,3H),6.84(d,J=8.5Hz,1H),6.41(s,2H).

[0315] Step 3: Preparation of N-(4-benzoyl-2-chlorophenyl)acrylamide (Example 29)

[0316] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.50 g, 68%).

[0317] EI-MS: [M] + 285.10. 1 H NMR(400MHz, DMSO-d6)δ9.90(s,1H),8.12(d,J=8.5Hz,1H),7.80(d,J=1.9Hz,1H),7.77-7.62(m,4H),7.55( t,J=7.5Hz,2H),6.71(dd,J=16.9,10.3Hz,1H),6.30(dd,J=17.0,1.8Hz,1H),5.82(dd,J=10.2,1.8Hz,1H).

[0318] Example 30

[0319]

[0320] Step 1: Preparation of (4-tert-butylphenyl)(3-chloro-4-hydroxyphenyl) methyl ketone (intermediate 35-b)

[0321] The operation steps are the same as the second step in the preparation of Example 1, and the product is a white solid (2.50 g, 60%).

[0322] EI-MS: [M] + 288.10. 1 H NMR (400MHz, DMSO-d6) δ11.25(s,1H),7.72(d,J=2.2Hz,1H),7.65(d,J=8.4Hz,2H),7.60-7.54(m,3H),7.10(d,J=8.6Hz,1H),1.33(s,9H).

[0323] Step 2: Preparation of (4-amino-3-chlorophenyl)(4-tert-butylphenyl) methyl ketone (intermediate 35-c)

[0324] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (1.11 g, 44%).

[0325] EI-MS: [M] + 287.15. 1 H NMR (400MHz, DMSO-d6) δ7.62-7.57(m,3H),7.57-7.53(m,2H),7.49(dd,J=8.5,2.0Hz,1H),6.84(d,J=8.5Hz,1H),6.36(s,2H),1.33(s,9H).

[0326] Step 3: Preparation of N-(4-(4-(tert-butyl)benzoyl)-2-chlorophenyl)acrylamide (Example 30)

[0327] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.91 g, 55%).

[0328] EI-MS: [M] + 341.10. 1 H NMR (400MHz, DMSO-d6) δ9.93(s,1H),8.13(d,J=8.5Hz,1H),7.83(d,J=2.0Hz,1H),7.76-7.67(m,3H),7.60(d,J=8 .6Hz,2H),6.74(dd,J=17.0,10.2Hz,1H),6.34(dd,J=17.0,1.9Hz,1H),5.85(dd,J=10.2,1.9Hz,1H),1.33(s,9H).

[0329] Example 31

[0330]

[0331] Step 1: Preparation of (3-chloro-4-hydroxyphenyl)(naphth-2-yl)methyl ketone (intermediate 36-b)

[0332] The operation steps are the same as the second step in the preparation of Example 1, and the product is a white solid (3.20 g, 80%).

[0333] EI-MS: [M] + 282.05. 1 H NMR (400MHz, DMSO-d6) δ11.31(s,1H),8.37-8.27(m,1H),8.23-7.98(m,3H),7.88-7.59(m,5H),7.25-7.06(m,1H).

[0334] Step 2: Preparation of (4-amino-3-chlorophenyl)(naphth-2-yl)methyl ketone (intermediate 36-c)

[0335] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (0.43 g, 72%).

[0336] EI-MS: [M] + 281.10. 1 H NMR (400MHz, DMSO-d6) δ8.23(s,1H),8.09(dd,J=8.1,1.5Hz,1H),8.07-7.97(m,2H),7.74(dd,J=8.5,1.7Hz,1H),7.68(d ,J=2.0Hz,1H),7.63(dddd,J=18.9,8.1,6.8,1.4Hz,2H),7.56(dd,J=8.5,2.0Hz,1H),6.87(d,J=8.6Hz,1H),6.41(s,2H).

[0337] Step 3: Preparation of N-(4-(2-naphthoyl)-2-chlorophenyl)acrylamide (Example 31)

[0338] The procedure was performed as described in step 2 of Example 25, and the product was a white solid (65 mg, 55%).

[0339] EI-MS: [M] + 335.05. 1H NMR(400MHz,DMSO-d6)δ9.94(s,1H),8.36(s,1H),8.21-8.02(m,4H),7.96-7.84(m,2H),7.79(dd,J=8.5,2.0Hz,1 H),7.73-7.60(m,2H),6.75(dd,J=17.0,10.2Hz,1H),6.35(dd,J=17.0,1.9Hz,1H),5.86(dd,J=10.2,1.9Hz,1H).

[0340] Example 32

[0341]

[0342] Step 1: Preparation of 2-chloro-4-(cyclohexylmethyl)aniline (intermediate 37-d)

[0343] The operation steps are the same as the third step in the preparation of Example 9, and the product is a white solid (1.20 g, 64%).

[0344] EI-MS: [M] + 223.10. 1 H NMR(400MHz,DMSO-d6)δ6.98-6.89(m,1H),6.87-6.78(m,1H),6.71-6.65(m,1H),5.08(s,2H),2.29(dd, J=7.0,3.2Hz,2H),1.59(d,J=12.5Hz,5H),1.38(s,1H),1.20-1.06(m,3H),0.86(q,J=11.5,10.3Hz,2H).

[0345] Step 2: Preparation of N-(2-chloro-4-(cyclohexylmethyl)phenyl)acrylamide (Example 32)

[0346] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.40 g, 31%).

[0347] EI-MS:[M]+277.15.1H NMR (400MHz, DMSO-d6) δ9.65(s,1H),7.62(d,J=8.1Hz,1H),7.28(s,1H),7.11(d,J=10.2Hz,1H),6.58(dd,J=17.0,10.1Hz,1H),6.42-6. 17(m,1H),5.84-5.60(m,1H),2.44(d,J=7.1Hz,2H),1.73-1.51(m,5H),1.48(t,J=9.1Hz,1H),1.13(d,J=9.5Hz,3H),1.00-0.80(m,2H).

[0348] Example 33

[0349]

[0350] Step 1: Preparation of 4-benzyl-2-chloroaniline (Intermediate 38-d)

[0351] The operation steps are the same as in step 3 of the preparation in Example 9, and the product obtained is a white solid (0.70 g, 26%).

[0352] EI-MS: [M] + 217.05. 1 H NMR (400MHz, DMSO-d6) δ7.27(t,J=7.6Hz,2H),7.18(dd,J=13.4,7.1Hz,3H),7.03(d,J=2 .0Hz,1H),6.89(dd,J=8.2,2.0Hz,1H),6.72(d,J=8.1Hz,1H),5.15(s,2H),3.76(s,2H).

[0353] Step 2: Preparation of N-(4-phenylmethyl-2-chlorophenyl)acrylamide (Example 33)

[0354] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.21 g, 57%).

[0355] EI-MS:[M]+271.10.1H NMR (400MHz, DMSO-d6) δ9.67(s,1H),7.64(d,J=8.3Hz,1H),7.37(d,J=2.0Hz,1H),7.32-7.23(m,4H),7.20(td,J=6.1,5 .5,2.4Hz,2H),6.58(dd,J=17.0,10.2Hz,1H),6.24(dd,J=17.0,2.0Hz,1H),5.76(dd,J=10.2,2.0Hz,1H),3.93(s,2H).

[0356] Example 34

[0357]

[0358] Step 1: Preparation of (4-amino-3-bromophenyl)(phenyl)methyl ketone (intermediate 39-c)

[0359] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (0.43 g, 43%).

[0360] EI-MS: [M] + 281.05. 1 H NMR (400MHz, DMSO-d6) δ7.92(d,J=2.0Hz,1H),7.70(dd,J=8.5,2.0Hz,1H),6.79(d,J=8.5 Hz,1H),6.21(s,2H),3.22(tt,J=11.0,3.2Hz,1H),1.83-1.55(m,5H),1.53-1.12(m,5H).

[0361] Step 2: Preparation of N-(2-chloro-4-(cyclohexylcarbonyl)phenyl)acrylamide (Example 34)

[0362] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.77 g, 60%).

[0363] EI-MS: [M] + 335.05. 1H NMR (400MHz, DMSO-d6) δ9.77(s,1H),8.13(d,J=1.7Hz,1H),7.99-7.88(m,2H),6.64(dd,J=17.0,10.2Hz ,1H),6.28(dd,J=16.9,1.9Hz,1H),5.80(dd,J=10.2,1.9Hz,1H),3.48-3.32(m,1H),1.77-1.61(m,5H).

[0364] Example 35

[0365]

[0366] Step 1: Preparation of (3-bromo-4-hydroxyphenyl)(phenyl)methyl ketone (intermediate 40-b)

[0367] The operation steps are the same as the second step in the preparation of Example 1, and the product is a white solid (1.20 g, 81%).

[0368] EI-MS: [M] + 275.95. 1 H NMR (400MHz, DMSO-d6) δ11.34 (s, 1H), 7.87 (d, J = 2.1Hz, 1H), 7.71-7.61

[0369] (m,4H),7.55(t,J=7.6Hz,2H),7.08(dd,J=8.5,1.4Hz,1H).

[0370] Step 2: Preparation of (4-amino-3-bromophenyl)(phenyl)methyl ketone (intermediate 40-c)

[0371] The procedure was performed as described in the first step of the preparation in Example 25, and the product was a white solid (0.33 g, 41%).

[0372] EI-MS: [M] + 274.95. 1 H NMR (400MHz, DMSO-d6) δ7.77(d,J=2.0Hz,1H),7.68-7.59(m,3H),7.56-7.46(m,3H),6.85(d,J=8.5Hz,1H),6.36(s,2H).

[0373] Step 3: Preparation of N-(4-benzoyl-2-bromophenyl)acrylamide (Example 35)

[0374] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.20 g, 57%).

[0375] EI-MS: [M] + 329.00. 1 H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 8.00 (s, 2H), 7.77-7.66 (m, 4H), 7.58 (t, J = 7.6Hz, 2H) ,6.69(dd,J=17.0,10.2Hz,1H), 6.33(dd,J=17.0,1.9Hz,1H), 5.85(dd,J=10.2,1.9Hz,1H).

[0376] Example 36

[0377]

[0378] Step 1: Preparation of (4-amino-3-bromophenyl)(phenyl)methyl ketone (intermediate 41-d)

[0379] Intermediate 39-C (0.20 g, 0.7 mmol), pinacol phenylboronic acid ester (0.17 g, 0.85 mmol), bis(dibenzylacetone)palladium (23 mg, 0.04 mmol), tricyclohexylphosphine (31 mg, 0.11 mmol), and potassium carbonate (0.29 g, 2.10 mmol) were weighed and added sequentially to a reaction flask. Under N2 protection, 1,4-dioxane (5 mL) and H2O (1 mL) were added, and the mixture was heated to reflux for 2 h. After cooling to room temperature, the mixture was extracted with ethyl acetate, and the organic phases were combined. The mixture was back-extracted with saturated brine, dried over anhydrous sodium sulfate, and filtered. Separation by silica gel column chromatography yielded a white solid (0.14 g, 70%).

[0380] EI-MS: [M] + 279.10. 1 H NMR (400MHz, DMSO-d6) δ7.71 (dd, J=8.3, 1.8Hz, 1H), 7.58 (d, J=2.1Hz, 1H), 7.55-7.34 (m, 5H), 6.7 9(d,J=8.5Hz,1H),5.68(s,2H),3.27(t,J=10.9Hz,1H),1.72(d,J=9.8Hz,5H),1.46-1.15(m,5H).

[0381] Step 2: Preparation of N-(5-(cyclohexylcarbonyl)-[1,1'-biphenyl]-2-yl)acrylamide (Example 36)

[0382] The procedure was performed as described in step 2 of Example 25, and the product was a white solid (30 mg, 32%).

[0383] EI-MS: [M] + 333.15. 1 H NMR (400MHz, DMSO-d6) δ9.60 (s, 1H), 7.98 (d, J = 2.1Hz, 1H), 7.85 (d, J = 9.4Hz, 2H), 7.51-7.44 (m, 2H), 7.41 (d, J = 7.3Hz, 3H), 6.38 (dd, J = 1 7.0,10.1Hz,1H),6.20(dd,J=17.1,2.0Hz,1H),5.71(dd,J=10.1,2.1Hz,1H),3.44(d,J=7.6Hz,1H),1.86-1.63(m,5H),1.42-1.14(m,5H).

[0384] Example 37

[0385]

[0386] Step 1: Preparation of (4-amino-3-(1H-pyrazol-3-yl)phenyl)(cyclohexyl) ketone (intermediate 42-d)

[0387] The procedure was performed as described in the first step of the preparation in Example 35, and the product was a white solid (90 mg, 11%).

[0388] EI-MS: [M] + 269.15. 1 H NMR (400MHz, CDCl3) δ8.25(d,J=2.1Hz,1H),7.74(dd,J=8.5,2.1Hz,1H),7.65(d,J=2.5Hz,1H),6.80(d, J=2.5Hz,1H),6.72(d,J=8.5Hz,1H),3.24(tt,J=11.6,3.2Hz,1H),1.93-1.69(m,5H),1.30-1.24(m,5H).

[0389] Step 2: Preparation of N-(4-(cyclohexylcarbonyl)-2-(1H-pyrazol-3-yl)phenyl)acrylamide (Example 37)

[0390] The procedure was performed as described in step 2 of Example 25, and the product was a white solid (15 mg, 8%).

[0391] EI-MS: [M] + 323.15. 1H NMR (400MHz, DMSO-d6) δ14.24(s,1H),13.02(s,1H),9.54(d,J=8.6Hz,1H),9.15(d,J=2.1Hz,1H),8.87-8.65(m,2H),7.86(d,J= 2.6Hz,1H),7.24-7.16(m,2H),6.75(dd,J=7.7,3.8Hz,1H),4.32(tt,J=11.0,3.2Hz,1H),2.74-2.47(m,5H),2.34-2.03(m,5H).

[0392] Example 38

[0393]

[0394] Step 1: Preparation of (4-amino-3-(1-methyl-1H-pyrazole-3-yl)phenyl)(cyclohexyl) ketone (intermediate 43-d)

[0395] The procedure was performed as described in the first step of the preparation in Example 35, and the product was a white solid (0.12 g, 30%).

[0396] EI-MS: [M] + 283.20. 1 H NMR (400MHz, DMSO-d6) δ8.09(d,J=2.1Hz,1H),7.79(d,J=2.3Hz,1H),7.66(dd,J=8.6,2.1Hz,1H),7.15 (s,2H),6.83-6.74(m,2H),3.91(s,3H),3.33-3.26(m,1H),1.74(d,J=10.2Hz,5H),1.47-1.14(m,5H).

[0397] Step 2: Preparation of N-(4-(cyclohexylcarbonyl)-2-(1-methyl-1H-pyrazole-3-yl)phenyl)acrylamide (Example 38)

[0398] The procedure was performed as described in step 2 of Example 25, and the product was a white solid (12 mg, 9%).

[0399] EI-MS: [M] + 337.20. 1H NMR (400MHz, DMSO-d6) δ11.99(s,1H),8.72(d,J=8.8Hz,1H),8.29(d,J=2.1Hz,1H),7.99-7.89(m,2H),7.00(d,J=2.4Hz,1H),6.48(dd,J=17.0, 10.1Hz,1H),6.36(dd,J=17.1,1.5Hz,1H),5.92(dd,J=10.1,1.5Hz,1H),4.01(s,3H),3.53-3.43(m,1H),1.84-1.65(m,5H),1.43-1.13(m,5H).

[0400] Example 39

[0401]

[0402] Step 1: Preparation of (4-amino-3-(1-methyl-1H-pyrazole-5-yl)phenyl)(cyclohexyl) ketone (intermediate 44-d)

[0403] The procedure was performed as described in the first step of the preparation in Example 35, and the product was a white solid (0.30 g, 75%).

[0404] EI-MS: [M] + 283.15. 1 H NMR (400MHz, DMSO-d6) δ7.78(dd,J=8.6,2.1Hz,1H),7.62(d,J=2.1Hz,1H),7.52(d,J=1.9Hz,1H),6.79(s,1H), 6.31(d,J=1.9Hz,1H),5.82(s,2H),3.64(s,3H),3.27(t,J=11.2Hz,1H),1.80-1.62(m,5H),1.42-1.11(m,5H).

[0405] Step 2: Preparation of N-(4-(cyclohexylcarbonyl)-2-(1-methyl-1H-pyrazole-5-yl)phenyl)acrylamide (Example 39)

[0406] The procedure was performed as described in step 2 of Example 25, and the product was a white solid (24 mg, 20%).

[0407] EI-MS: [M] + 337.15. 1H NMR (400MHz, DMSO-d6) δ9.57 (s, 1H), 8.07-7.97 (m, 2H), 7.89 (dd, J = 5.5, 2.0Hz, 1H) ,7.53(d,J=2.0Hz,1H),6.43(dd,J=17.0,10.2Hz,1H),6.34(dd,J=4.9,1.9Hz,1H),6 .22(dd,J=17.0,2.0Hz,1H),5.75(dd,J=10.2,2.0Hz,1H),3.64(s,3H),3.44(ddt,J= 11.0, 6.6, 3.3Hz, 1H), 1.76 (dd, J=25.6, 12.4Hz, 5H), 1.36 (dq, J=26.2, 12.4Hz, 5H).

[0408] Example 40

[0409]

[0410] Step 1: Preparation of (4-amino-3-(pyridin-3-yl)phenyl)(cyclohexyl) methyl ketone (intermediate 45-d)

[0411] The procedure was performed as described in the first step of the preparation in Example 35, and the product was a white solid (0.42 g, 84%).

[0412] EI-MS: [M] + 280.15. 1 H NMR (400MHz, DMSO-d6) δ8.64-8.52(m,2H),7.84(dt,J=7.8,1.9Hz,1H),7.75(dd,J=8.5,2.1Hz,1H),7.61(d,J=2.0Hz,1H),7.49( dd,J=7.9,4.7Hz,1H),6.81(d,J=8.5Hz,1H),5.85(s,2H),3.29(td,J=9.7,8.2,5.7Hz,1H),1.80-1.60(m,5H),1.50-1.11(m,5H).

[0413] Step 2: Preparation of N-(4-(cyclohexylcarbonyl)-2-(pyridin-3-yl)phenyl)acrylamide (Example 40)

[0414] The operation steps are the same as the second step in the preparation of Example 25, and the product is a white solid (0.13 g, 36%).

[0415] EI-MS: [M] + 334.20. 1H NMR(400MHz,DMSO-d6)δ9.74(s,1H),9.78(s,1H),8.62-8.57(m,2H),8.03(dd,J=8.5,2.1 Hz,1H),7.91(d,J=2.1Hz,1H),7.86(d,J=8.5Hz,1H),7.82(dt,J=7.9,2.0Hz,1H),7.49(d d,J=8.3,5.2Hz,1H),6.37(dd,J=17.0,10.1Hz,1H),6.19(dd,J=17.0,2.0Hz,1H),5.72(d d,J=10.1,2.1Hz,1H),3.48(td,J=10.9,3.3Hz,1H),1.86-1.64(m,5H),1.49-1.28(m,5H).

[0416] Table 2. Names of products obtained in Examples 1-40

[0417]

[0418]

[0419] To determine the affinity of the 2-phenoxyacetic acid derivatives in Examples 1-24 of this invention for the protein TEAD, a thermal shift assay (TSA; Experiment 1) and an isothermal titration calorimetry (ITC; Experiment 2) were performed, as detailed below:

[0420] Experiment 1: Protein Thermal Migration Experiment

[0421] 1.1 Experimental Objective: To determine the effect of the compounds in Examples 1-24 on the thermal stability of TEAD using protein thermal migration experiments, and to determine the effect based on ΔT m The affinity of the compound for TEAD was evaluated.

[0422] 1.2 Experimental instruments: The CFX Connect real-time PCR system was purchased from Bio-Rad, and the pipettes were purchased from Eppendorf.

[0423] 1.3 Experimental Methods: The effect of the compounds in the examples on the thermal stability of TEAD was detected using a protein thermal migration assay. The thermal denaturation of the protein during programmed temperature rise was monitored in real time using a Sypro Orange fluorescent probe, and the thermal denaturation temperature Ta of the protein was calculated by fitting the data. m After treatment with compounds, T m Change ΔT m Value. The specific experimental procedure is as follows:

[0424] Each 20 μL experimental volume contained purified TEAD2_C380S protein solution (final concentration: 10 μmol / L), the test compound (final concentration: 200 μmol / L), a 5×Sypro Orange fluorescent probe, and a buffer system consisting of 50 mmol / L KH₂PO₄, 150 mmol / L NaCl, 0.2 mmol / L TCEP, and 2% DMSO (pH 7.4). Experiments were performed on a CFX Connect real-time PCR system, with samples heated from 20 °C to 95 °C at a rate of 0.5 °C / s.

[0425] 1.4 Experimental Data Processing Methods: Based on the fluorescence-temperature curves automatically recorded by the CFX Connect real-time PCR system, the negative derivative curve of the fluorescence-temperature curve was automatically generated using the accompanying analysis software (Bio-Rad CFX Maestro). The inflection point of this curve is the protein's thermal denaturation temperature T. m And calculate T after compound treatment. m Change ΔT m value.

[0426] 1.5 Experimental Conclusions:

[0427] Experimental results show that the compounds in Examples 1-24 of this invention have a significant effect on the thermal stability of TEAD, proving that the compounds have a strong interaction with TEAD. The compounds cause a change in the thermal denaturation temperature of TEAD by ΔT. m The data is shown in Table 3 below.

[0428] Table 3. Effects of compounds from Examples 1-24 and existing TEAD inhibitors on the thermal stability of TEAD.

[0429]

[0430]

[0431] Experiment 2: Isothermal titration calorimetry experiment

[0432] 2.1 Experimental Objective: To determine the interaction reaction parameters between the compounds in Examples 1-24 and TEAD, based on the dissociation constant K. d Evaluate the affinity of the compound for TEAD.

[0433] 2.2 Experimental Apparatus: The isothermal titration calorimeter (PEAQ-ITC) was purchased from Malvern, and the pipettes were purchased from Eppendorf.

[0434] 2.3 Experimental Methods: The affinity of the compounds in the examples for TEAD was determined using isothermal titration calorimetry. The heat change generated by the binding of the compounds with the protein was measured using an isothermal titration calorimeter. Various reaction parameters of the interaction between the compounds and TEAD were calculated using fitting, based on the dissociation constant K. d The affinity of the compound for TEAD was characterized. The specific experimental procedures are as follows:

[0435] ITC experiments were performed on an isothermal titration calorimeter. The buffer system consisted of 50 mmol / L KH₂PO₄, 150 mmol / L NaCl, 0.2 mmol / L TCEP, and 2% DMSO (pH 7.4). The purified TEAD2_C380S protein solution was loaded into the calorimeter's calorimeter to a final concentration of 100 μmol / L. The same buffer system was automatically loaded into the titration syringe to a final concentration of 2 mmol / L. The experiment was conducted according to the instrument's program: 19 titrations, 2 μL each, stirring at 750 rpm, at 25 °C.

[0436] 2.4 Experimental Data Processing Methods: The experimental data were analyzed using the single binding site model. The reaction parameters of the interaction were calculated using the accompanying analysis software (MicroCal PEAQ-ITC Analysis Soltware) of the isothermal titration calorimeter, and the dissociation constant K was determined. d .

[0437] 2.5 Experimental Conclusions:

[0438] The above methods demonstrate that the compounds in Examples 1-24 of this invention have a high affinity for TEAD, and their dissociation constant K d The data is shown in Table 4 below.

[0439] Table 4 shows the compounds in Examples 1-24 and the dissociation constant K of TEAD for existing TEAD inhibitors. d

[0440]

[0441]

[0442] Furthermore, to verify the inhibitory effect of the compounds in the embodiments of the present invention on TEAD transcriptional activity, relevant tests were conducted, and the specific test procedures are as follows:

[0443] Experimental objective: To test the inhibitory effect of the compounds in the examples on TEAD transcriptional activity.

[0444] Experimental instruments: centrifuge purchased from Eppendorf, CO2 incubator purchased from Thermo, biosafety cabinet purchased from Shanghai Boxun Company, pipettes purchased from Eppendorf, and ELISA reader purchased from BioTek.

[0445] Experimental Methods: A dual-luciferase reporter assay was used to test the inhibitory effect of the compounds in this study on TEAD transcriptional activity in HEK293T cells. HEK293T cells were transfected with 8×GTIIC-luciferase plasmid and pRL-CMV, which express firefly luciferase and kidney luciferase, respectively. The relative fluorescence intensity was determined by measuring the fluorescence intensity of firefly and kidney luciferase and calculating their ratio to characterize TEAD transcriptional activity. The inhibition rate of different compounds on TEAD transcriptional activity was also calculated.

[0446] The specific experimental procedure is as follows:

[0447] HEK293T cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin at 37°C and 5% CO2. HEK293T cells were seeded at a density of 3000 cells / well in 96-well plates and cultured overnight at 37°C and 5% CO2. After cell attachment, the medium was discarded, and 50 ng of 8×GTIIC-luciferase plasmid and 0.5 ng of pRL-CMV plasmid were transfected into HEK293T cells using Lipofectamine 2000. After 4 h of transfection at 37°C and 5% CO2, the transfection reagents were discarded, and 50 μmol / L or 5 μmol / L solutions of the respective compounds were added. A positive control (0.1‰ DMSO) and a negative control (no plasmid transfection) were also included, and the cells were cultured for another 24 h at 37°C and 5% CO2. After discarding the culture medium, the fluorescence intensity of fireflies and sea urchins was measured on a microplate reader using the Dual-Luciferase Reporter Assay kit. The ratio between the two was calculated, and the inhibition rate of different compounds on TEAD transcriptional activity was further calculated.

[0448] Experimental data processing methods:

[0449] The percentage inhibition rate of the test compound on TEAD transcriptional activity was calculated using the following formula:

[0450] Relative fluorescence intensity = (fluorescence intensity of fireflies in the test group - fluorescence intensity of fireflies in the negative control group) / (fluorescence intensity of sea snails in the test group - fluorescence intensity of sea snails in the negative control group);

[0451] % Inhibition rate = (1 - relative fluorescence intensity of test compound / relative fluorescence intensity of positive control) × 100%.

[0452] Experimental conclusion:

[0453] The above methods demonstrate that the compounds in the embodiments of this invention have a significant inhibitory effect on TEAD transcriptional activity. The percentage inhibition rate of TEAD transcriptional activity at concentrations of 50 μmol / L or 5 μmol / L is shown in Table 5 below.

[0454] Table 5. Inhibition rates (%) of TEAD transcriptional activity by the compounds in Examples 1–40 and existing TEAD inhibitors.

[0455]

[0456] Furthermore, to verify the inhibitory effect of the N-phenylacrylamide derivatives in Examples 25-40 of this invention on the proliferation of NF2-deficient lung cancer cells NCI-H226, relevant tests were conducted, and the specific test procedures are as follows:

[0457] Experimental objective: To test the inhibitory activity of the compounds in Examples 25-40 on the proliferation of NF2-deficient lung cancer cells NCI-H226.

[0458] Experimental instruments: centrifuge purchased from Eppendorf, CO2 incubator purchased from Thermo, biosafety cabinet purchased from Shanghai Boxun Company, pipettes purchased from Eppendorf, and ELISA reader purchased from BioTek.

[0459] Experimental Methods: The inhibitory effect of the compounds in the examples on the proliferation of NF2-deficient lung cancer cells NCI-H226 was detected using the CCK-8 assay. NCI-H226 cells were cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin-streptomycin at 37°C and 5% CO2. NCI-H226 cells were seeded at a density of 3000 cells / well in 96-well plates and cultured overnight at 37°C and 5% CO2. After cell attachment, different concentrations of the compound solution were added, along with negative and blank controls. The cells were cultured for another 4 days at 37°C and 5% CO2. Then, 20 μL of CCK-8 reagent was added to each well, vortexed, and cultured for another 1.5 h. After CCK-8 staining, the 96-well plates were placed on a microplate reader, and the absorbance was measured at 450 nm to calculate the cell proliferation inhibition rate.

[0460] Experimental data processing methods:

[0461] The percentage inhibition rate of the compounds in the examples on the proliferation of NCI-H226 cells was calculated using the following formula:

[0462] % Inhibition rate = [1 – (Test compound value – Blank control value) / (Negative control value – Blank control value)] × 100%

[0463] The IC was calculated using a four-parameter nonlinear logic formula fitted with data of different concentrations and corresponding percentage inhibition rates using GraphPad Prism 8.0. 50 value.

[0464] Experimental conclusion:

[0465] The above methods demonstrate that compounds 25-40 of the embodiments shown in this invention have significant inhibitory activity against the proliferation of NF2-deficient lung cancer cells NCI-H226, with an IC50 value of [missing information]. 50 The values ​​are shown in Table 6 below.

[0466] Table 6. Inhibitory activity (IC50) of compounds in Examples 25-40 against NCI-H226 proliferation (IC50). 50 μmol / L, n=3)

[0467] Example <![CDATA[IC 50 ]]> Example <![CDATA[IC 50 ]]> Example 25 22.60±3.75 Example 33 5.32±2.90 Example 26 6.62±2.06 Example 34 6.12±0.76 Example 27 1.24±0.15 Example 35 7.01±3.55 Example 28 4.81±1.12 Example 36 2.18±0.53 Example 29 5.70±1.28 Example 37 0.61±0.09 Example 30 3.66±0.77 Example 38 1.94±0.55 Example 31 1.55±0.41 Example 39 0.34±0.02 Example 32 16.90±2.61 Example 40 1.42±0.23

[0468] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

[0469] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A TEAD inhibitor, characterized in that, This includes compounds represented by general formula (I), their stereoisomers or salts thereof, or compounds represented by general formula (II), their stereoisomers or salts thereof: Wherein, the R1 group is a substituted alkyl or substituted acyl group; the R2 group is one of H, methyl, ethyl and benzyl; X1 is any one of H, Cl and Br, and X2 is H or Cl; Wherein, the R3 group is a substituted alkyl or substituted acyl group; the X3 is H, Cl, Br, Any one of them; X4 is H or Cl.

2. The TEAD inhibitor according to claim 1, characterized in that, The R1 group is a cycloalkyl-substituted alkyl group, an aromatic ring-substituted alkyl group, an alkyl-substituted acyl group, or an aromatic ring-substituted acyl group.

3. A TEAD inhibitor according to claim 2, characterized in that, The R1 base is: Any one of them.

4. A TEAD inhibitor according to claim 2, characterized in that, The R1 base is Any one of them.

5. A TEAD inhibitor according to claim 1, characterized in that, The R3 group is a cycloalkyl-substituted alkyl group, an aromatic ring-substituted alkyl group, a cycloalkyl-substituted acyl group, or an aromatic ring-substituted acyl group.

6. A TEAD inhibitor according to claim 5, characterized in that, The R3 base is 7. A TEAD inhibitor according to claim 5, characterized in that, The R3 base is Any one of them.

8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a TEAD inhibitor as described in any one of claims 1 to 7, and one or more pharmaceutically acceptable carriers, diluents, or excipients; the pharmaceutical composition is prepared into tablets, capsules, soft capsules, or injections by adding excipients to a TEAD inhibitor as described in any one of claims 1 to 7; the excipients include one or more of additives, stabilizers, solubilizers, lubricants, and disintegrants.

9. The use of a TEAD inhibitor according to any one of claims 1 to 7 in the preparation of a TEAD inhibitor drug or a drug for inhibiting TEAD-YAP protein-protein interactions.

10. The use of a TEAD inhibitor according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 8 in the preparation of a cancer drug; characterized in that, The cancers mentioned include any one of the following: breast cancer, pancreatic cancer, non-small cell lung cancer, thyroid cancer, seminoma, melanoma, bladder cancer, liver cancer, stomach cancer, colorectal cancer, kidney cancer, ovarian cancer, uterine cancer, prostate cancer, glioma, myelodysplastic syndrome, and acute myeloid leukemia.

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