A kind of aryloxyaminobenzoic acid derivative and its preparation method and application

Abstract

The present invention belongs to the field of derivatives and their synthesis, and specifically discloses an aryloxyaminobenzoic acid derivative and a preparation method and application thereof. A derivative as shown in formula (I) is provided, wherein a nitrobenzoic acid compound is used as a raw material, and the aryloxyaminobenzoic acid derivative is prepared through a multi-step chemical reaction such as amidation, reduction, and acidification. In addition, the present invention also discloses the derivative shown in formula (I) or a pharmaceutically reducible salt thereof, or a pharmaceutical composition thereof as a farnesoid X receptor (FXR) modulator, for the development of drugs for preventing and / or treating diseases mediated and / or involved in FXR.

Description

Technical Field

[0001] The invention belongs to the field of derivatives and their synthesis, and specifically relates to an aryloxyaminobenzoic acid derivative and a preparation method and application thereof. Background Art

[0002] Farnesoid X receptor (FXR), also known as bile acid receptor, is a member of the nuclear receptor superfamily and is mainly distributed in the liver, intestines and other organs. FXR plays a regulatory role in multiple metabolic pathways and plays an important role in bile acid, cholesterol, lipid and glucose metabolism. In addition, FXR is closely related to cholestasis, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, gallstones, diabetes, atherosclerosis and cancer.

[0003] Non-alcoholic fatty liver disease (NAFLD) has a complex pathogenesis and is currently the most common chronic liver disease in the world. Non-alcoholic steatohepatitis (NASH) is a progressive stage of NAFLD and a liver manifestation of metabolic syndrome. As a key metabolic regulator, FXR has become a very promising drug target for the treatment of chronic liver diseases such as NAFLD and NASH (Zhou S, et al. Biomed Pharmaco, 2022, 154: 113577; Panzitt K, et al. Mol Cell Endocrinol, 2022: 111678).

[0004] FXR modulators include FXR agonists and antagonists (inhibitors). Currently, many pharmaceutical companies and research institutes have invested a lot of manpower and material resources in the research and development of FXR agonists, and many different structural types of FXR agonists have been discovered and reported. FXR agonists can be generally divided into two categories: one is steroidal, represented by Intercept's obeticholic acid (OCA); the other is non-steroidal, represented by GlaxoSmithKline's GW4064. The bile acid-derived FXR agonist OCA has been approved for marketing by the US FDA in May 2016 as a treatment for primary biliary cholangitis. The Phase III clinical study of OCA for NASH indications has been completed. The results show that OCA is effective in treating NASH combined with liver fibrosis, but the results also indicate that it has a negative impact on blood lipids, including increasing low-density lipoprotein (LDL) and reducing high-density lipoprotein (HDL) levels. In patients with a higher risk of cardiovascular disease, these negative effects may bring major side effects, and some patients experience skin itching, with an incidence of not less than 5% and other safety issues. These adverse reactions will limit the clinical use of OCA in NASH. GW4064 is the first non-steroidal FXR agonist obtained by GlaxoSmithKline through high-throughput screening. It has high selectivity and strong in vitro activity. However, the bioavailability of this compound is very low, less than 10% in rats, so it has not entered the clinical development stage. Some other non-steroidal active compounds developed by other studies have good FXR target selectivity, but most of them have problems with poor solubility and pharmacokinetic properties.

[0005] Literature reports that aminobenzoic acid compounds are a class of FXR agonists, for example, Bioorganic & Medicinal Chemistry, 2015, 23: 499–514; Bioorganic & Medicinal Chemistry, 2014, 22(8): 2447-2460; Journal of medicinal chemistry, 2014, 57(19): 8035–8055; ChemMedChem, 2018, 13(23): 2530-2545.

[0006] Although the structural modification of aminobenzoic acid compounds has received widespread attention and a large number of aminobenzoic acid derivatives have been synthesized, there are still deficiencies in drugability such as biological activity, water solubility and oral bioavailability. So far, no aminobenzoic acid derivative has entered clinical trial research. Summary of the invention

[0007] In order to develop clinical drug resources, the present invention aims to provide a preparation method and application of aryloxyaminobenzoic acid derivatives. Different substituted phenol or naphthol derivatives are selected as raw materials during the preparation, and aryloxyaminobenzoic acid derivatives are prepared through multi-step chemical synthesis reactions.

[0008] In the first aspect of the present invention, there is provided an aryloxyaminobenzoic acid derivative as shown in the following formula (I):

[0009]

[0010] Wherein: Ar is a substituted phenyl or naphthyl, preferably selected from 4-ethylphenyl, 4-tert-butylphenyl, 2,4-di-tert-butylphenyl, 4-acetylphenyl, 4-benzyloxyphenyl, 4-propionylphenyl, biphenyl, naphthyl;

[0011] n is independently selected from 2 and 3;

[0012] R1 and R2 are each independently selected from hydrogen, methyl, and ethyl;

[0013] R1, R2 and the -COOH group are located at any position of the benzene ring.

[0014] In the first aspect of the present invention, the derivative represented by formula (I) is selected from the following compounds:

[0015]

[0016]

[0017] In the present invention, the pharmaceutically acceptable salt of the derivative shown in formula (I) is preferably a salt produced by the reaction of the carboxyl group of the present invention and a basic compound. The basic compound is preferably selected from sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate or potassium bicarbonate. The above pharmaceutically acceptable salt is easy to separate, the compound shown in formula (I) can be purified by column chromatography, and the salt of the derivative shown in formula (I) and the basic compound can be purified by recrystallization.

[0018] The second aspect of the present invention provides a method for preparing a derivative having a structure as shown in general formula (I), and the preparation route is as follows:

[0019]

[0020] The preparation method comprises the following steps:

[0021] Step a. Nitrobenzoic acid and its different derivatives II and ethyl aminobenzoate and its different derivatives III are subjected to an amidation reaction in a suitable solvent and a condensation reagent to prepare an intermediate compound IV;

[0022] Step b. The compound represented by formula IV is subjected to reduction reaction in an organic solvent and a reducing agent to obtain compounds 5a-f;

[0023] Step c. Compounds 5a-f are subjected to amidation reaction with aromatic oxygen carboxylic acid derivatives 6a-p respectively to generate aromatic oxygen aminobenzoic acid ester derivatives shown in formula VI;

[0024] Step d: The compound represented by formula VI is hydrolyzed in an organic solvent under alkaline conditions, and then subjected to an acidification reaction to prepare a derivative represented by formula (I).

[0025] The definitions of R1 and R2 in the compounds represented by formula (II), formula (III), formula (IV), compounds 5a-f, and formula (VI) and the compound represented by formula (I) are consistent. The definitions of Ar and n in the compounds represented by compounds 6a-p, the compounds represented by formula (VI) and the compound represented by formula (I) are consistent.

[0026] Furthermore, in the preparation method, the molar ratio of the nitrobenzoic acid and its different derivatives II to the ethyl aminobenzoate and its different derivatives III in step a is 1:1.1, and the reaction temperature is 30-45°C;

[0027] The appropriate solvent in step a is selected from one of N,N-dimethylformamide (DMF), tetrahydrofuran (THF), dichloromethane, chloroform, and dimethyl sulfoxide (DMSO);

[0028] The condensation reagent described in step a may have different condensation conditions, preferably selected from 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (abbreviated as: HATU), N,N-diisopropylethylamine (abbreviated as: DIPEA), DMF, and the reaction temperature is 30-45°C; 1-ethyl-3(3-dimethylpropylamine)carbodiimide hydrochloride (abbreviated as: EDCI·HCl), 1-hydroxybenzotriazole (abbreviated as: HOBT), DMF, and the reaction temperature is 30-45°C; dicyclohexylcarbodiimide as a condensation agent, tetrahydrofuran, and the reaction temperature is 30-45°C.

[0029] Furthermore, in the preparation method, the molar ratio of the compound represented by formula (IV) described in step b to the reducing agent is 1:5, and the reaction temperature is room temperature to 45°C;

[0030] The organic solvent in step b is selected from one of ethanol, methanol, tetrahydrofuran and dichloromethane;

[0031] The reducing agent described in step b is selected from zinc powder or iron powder;

[0032] In step b, those skilled in the art can make appropriate selections of reaction conditions. For example, a preferred operation implementation method may be: the compound represented by formula IV is reduced in ethanol with zinc powder to obtain derivatives 5a-f; or the compound represented by formula IV is hydrogenated and reduced in a hydrogen atmosphere with 5% Pd / C as a catalyst to prepare compounds 5a-f.

[0033] Further, in the preparation method, the molar ratio of the compound 5a-f to the compound 6a-p in step c is 1:1.3;

[0034] Compounds 6a-p can be purchased or prepared;

[0035] The method of step c is the same as that of step a, and those skilled in the art can make appropriate selections of reaction conditions.

[0036] Furthermore, in the preparation method, the molar ratio of the compound represented by formula (VI) in step d to the base is 1:1.1;

[0037] The organic solvent in step d is selected from ethanol, methanol and tetrahydrofuran;

[0038] The base described in step d is selected from sodium hydroxide or potassium hydroxide;

[0039] The acidification reaction conditions described in step d can be appropriately selected by those skilled in the art. Preferably, 1 mol / L hydrochloric acid solution is used to adjust the pH to 1-2 and the reaction is carried out at 0-5°C.

[0040] In the third aspect of the present invention, the prepared derivative or its pharmaceutically acceptable salt, or its pharmaceutical composition is used as an FXR modulator for FXR-related diseases, such as non-alcoholic fatty liver disease, non-alcoholic fatty hepatitis, cholestatic liver disease, hyperlipidemia, metabolic diseases such as diabetes and complications, etc. The present invention first screens the synthesized aryloxyaminobenzoic acid derivatives for FXR agonist activity and cytotoxic activity by dual luciferase reporter gene assay, screens out the preferred compounds, and studies the improvement of free fatty acid-induced HepG-2 cell steatosis in vitro, and studies the pharmacodynamics of high-fat diet-induced NASH mice in vivo, as well as in vivo pharmacokinetics and acute toxicity studies. Studies have shown that the aryloxyaminobenzoic acid derivatives of the present invention can be used as non-steroidal FXR modulators, and such compounds can effectively excite FXR receptors, effectively improve dyslipidemia in high-fat diet-induced NASH mice, and have improved pharmacological activity, physicochemical properties and pharmacokinetic properties compared with compounds reported in the art. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1Schematic diagram of the improvement effect of Example compounds Z4 and Z5 on steatosis HepG-2;

[0042] (A) Schematic diagram of the cytotoxic effects of different concentrations of Z4 and Z5 on HepG-2 cells;

[0043] (B) Schematic diagram of the lipid toxicity effects of different concentrations of Z4 and Z5 on HepG-2 cells;

[0044] (C) Schematic diagram of the effects of Z4 and Z5 on the TG content of HepG-2 cells induced by FFA;

[0045] (D) Effects of Z4 and Z5 on FFA-induced fatty degeneration of HepG-2 cells (Oil Red O staining, light microscope 20 times, scale bar, 50 μm); (E) Schematic diagram of the effects of Z4 and Z5 on the absorbance of lipid droplets in FFA-induced lipid accumulation in HepG-2 cells;

[0046] Figure 2 Schematic diagram of the effect of Example Compound Z4 on obesity in NASH mice induced by high-fat diet and CCl4;

[0047] Wherein (A) Schematic diagram of modeling and treatment methods; (B) Body weight and weight change (n=6-8); (C) Fat / body weight ratio (n=8); (D) Adipocyte cross-sectional area (n=8); (E) From top to bottom, macroscopic images of perirenal white adipose tissue, macroscopic images of epididymal white adipose tissue, H&E staining of perirenal white adipose tissue sections, and H&E staining of epididymal white adipose tissue sections;

[0048] Figure 3 Schematic diagram showing the effect of Example Compound Z4 on liver steatosis and lipotoxicity in NASH mice;

[0049] (A) From top to bottom, macroscopic images of the livers of mice in the NCD, NASH, Z4 and OCA groups, and representative H&E images of liver sections; (B) liver weight (n=8); (C) liver / body weight ratio (%) (n=8); (D) liver TG content (n=8); (E) serum AST level (n=8); (F) serum ALT level (n=8); (G) liver CAT content (n=8); (H) serum GSH-Px content (n=8); (I) liver MDA content (n=8); (J) serum MDA content (n=8);

[0050] Figure 4 Schematic diagram of the effect of Example Compound Z4 on lipid metabolism disorders in NASH mice;

[0051] Among them, (A) serum TG content (n=8); (B) serum TC content (n=8); (C) serum HDL-C content (n=8); (D) serum LDL-C content (n=8);

[0052] Figure 5 Schematic diagram of the pharmacokinetics of Example Compound Z4 in C57BL6J mice;

[0053] Among them (A) pharmacokinetic parameters of Z4;

[0054] (B) Tissue distribution of Z4 in plasma, liver, and intestine of C57BL6J mice after a single oral administration (10 mg / kg) (n=3). DETAILED DESCRIPTION

[0055] The present invention is further described below by way of embodiments in combination with the accompanying drawings, but is not intended to limit the protection scope of the present invention.

[0056] Example 1: Preparation of Compounds 5a-f

[0057] 1. Preparation of ethyl 3-(2-aminobenzamido)benzoate (5a)

[0058] Add compound 2-nitrobenzoic acid (30mmol) to a reaction flask, add THF (40mL), HATU (36mmol), DIPEA (36mmol), and then add compound 3-aminobenzoic acid ethyl ester (33mmol). Heat the reaction solution to 30-45°C, react for 6-8h, and monitor by TLC (petroleum ether: ethyl acetate = 6:1-2:1, v / v). After the reaction is complete, remove the solvent under reduced pressure, add 300mL of water, extract with ethyl acetate (100mL×3), combine the organic phases, wash with saturated NaCl (100mL), add anhydrous MgSO4 to dry, filter, remove the solvent under reduced pressure, and dry at 35°C for 4h to obtain intermediate IV. Compound IV does not need to be purified and can be directly used in the next step.

[0059] Zinc powder (150mmol) was added to the reaction flask, anhydrous ethanol (100mL) was added, stirred, concentrated HCl (0.2eq) was slowly added dropwise, and stirred for activation for 5min. Compound IV (30mmol) was added, and the mixture was reacted at room temperature for 1h, and the temperature was raised to 45°C. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 5:1 to 1:1, v / v). After the reaction was completed, the solvent was concentrated under reduced pressure, water (300mL) was added, and the mixture was extracted with ethyl acetate (100mL×3), the organic phases were combined, washed with saturated NaCl (50mL×2), dried with anhydrous MgSO4, filtered, concentrated under reduced pressure, and dried at 35°C for 4h to obtain compound 5a.

[0060] The physicochemical and spectral data of compound 5a are as follows:

[0061]

[0062] Yellow solid, yield 68%, 1 H NMR (600MHz, DMSO-d6) δ10.88(s,1H),8.34(t,J=4.7Hz,1H),8.17(d,J=8.2Hz,1H),7.96–7.85 (m,2H),7.85–7.68(m,3H),7.52(t,J=7.9Hz,1H),4.33(q,J=7.1Hz,2H),1.33(t,J=7.1Hz,3H).

[0063] 2. The preparation methods, physicochemical and spectral data of compounds 5b~5f are as follows:

[0064] Ethyl 4-(2-aminobenzamido)benzoate (5b)

[0065]

[0066] According to the preparation method of compound 5a, 3-aminobenzoic acid ethyl ester was replaced by 4-aminobenzoic acid ethyl ester, and the other conditions were the same. Compound 5b: white solid, yield 65%, 1 H NMR(600MHz,DMSO-d6)δ11.00(s,1H),8.18(d,J=8.2Hz,1H),7.97(d,J=8.6Hz,2H),7 .89(d,J=7.5Hz,1H),7.84–7.74(m,4H),4.31(q,J=7.1Hz,2H),1.32(t,J=7.1Hz,3H).

[0067] Ethyl 2-(2-aminobenzamido)benzoate (5c)

[0068]

[0069] According to the preparation method of compound 5a, 3-aminobenzoic acid ethyl ester was replaced by 2-aminobenzoic acid ethyl ester, and the other conditions were the same. Compound 5c: yellow solid, yield 73%, 1H NMR (600MHz, DMSO-d6) δ11.39(s,1H),8.50(d,J=8.3Hz,1H),8.00(d,J=7.9Hz,1H),7.63(dd,J=18.2,7.8Hz,2H),7.26 -7.19(m,1H),6.80(d,J=8.2Hz,1H),6.64(t,J=7.5Hz,1H),6.57(s,1H),4.33(q,J=7.1Hz,2H),1.31(t,J=7.1Hz,3H).

[0070] Ethyl 2-(2-amino-5-methylbenzamide)benzoate (5d)

[0071]

[0072] According to the preparation method of compound 5a, 2-nitrobenzoic acid is replaced by 5-methyl-2-nitrobenzoic acid, 3-aminobenzoic acid ethyl ester is replaced by 2-aminobenzoic acid ethyl ester, and the other conditions are the same. Compound 5d: 1 H NMR (600MHz, DMSO-d6) δ11.25(s,1H),8.43(d,J=8.4Hz,1H),7.99(dd,J=7.9,1.4Hz,1H),7.72–7.56(m,1H),7.42(s,1H),7.28–7.1 6(m,1H),7.08(dd,J=8.3,1.5Hz,1H),6.72(d,J=8.4Hz,1H),6.33(s,2H),4.34(d,J=7.1Hz,2H),2.22(s,3H),1.31(t,J=7.1Hz,3H).

[0073] Ethyl 2-(2-amino-4-methylbenzamido)benzoate (5e)

[0074]

[0075] According to the preparation method of compound 5a, 2-nitrobenzoic acid was replaced by 4-methyl-2-nitrobenzoic acid, 3-aminobenzoic acid ethyl ester was replaced by 2-amino-5-methylbenzoic acid ethyl ester, and the other conditions were the same. Compound 5e: yellow solid, yield 78%, 1H NMR (600MHz, DMSO-d6) δ11.25(s,1H),8.39(d,J=8.5Hz,1H),7.80(s,1H),7.49(d,J=8.1Hz,1H),7.47–7.40(m,1H),6. 59(s,1H),6.54(s,2H),6.46(d,J=7.8Hz,1H),4.33(q,J=7.1Hz,2H),2.32(s,3H),2.21(s,3H),1.31(t,J=7.1Hz,3H).

[0076] Ethyl 3-(2-aminobenzamido)-4-ethylbenzoate (5f)

[0077]

[0078] According to the preparation method of compound 5a, 3-aminobenzoic acid ethyl ester was replaced by 3-amino-4-ethylbenzoic acid ethyl ester, and the other conditions were the same. Compound 5f: yellow solid, yield 77%, 1 H NMR (600MHz, DMSO-d6) δ9.76 (s, 1H), 7.88 (d, J = 1.4Hz, 1H), 7.80 (dd, J = 8.0, 1.6Hz,1H),7.72(d,J=7.9Hz,1H),7.44(d,J=8.0Hz,1H),7.21(t,J=7.7Hz,1 H),6.76(d,J=8.3Hz,1H),6.60(t,J=7.5Hz,1H),6.42(s,1H),4.32(q,J=7.1 Hz, 1H), 2.69 (q, J = 7.5Hz, 1H), 1.32 (t, J = 7.1Hz, 1H), 1.15 (t, J = 7.5Hz, 1H).

[0079] Example 2: Preparation of target compounds Z1-Z53

[0080] 1. Preparation of 3-(2-(4-(4-ethylphenoxy)butyrylamino)benzylamino)benzoic acid (Z1)

[0081] Compound 4-(4-ethylphenoxy)butyric acid (6a, 13mmol) was added to a reaction flask, followed by THF (16mL), HATU (15mmol), DIPEA (15mmol), and then compound 3-(2-aminobenzamido)ethyl benzoate (5a, 10mmol). The temperature was raised to 45-50°C, and the reaction was continued for 6-12h, and monitored by TLC (petroleum ether: ethyl acetate = 5:1-2:1, v / v). After the reaction was completed, the solvent was evaporated under reduced pressure, 120mL of water was added to the residue, and the mixture was extracted with ethyl acetate (40mL×3), the organic phases were combined, washed with saturated NaCl (40mL), dried with anhydrous MgSO4, filtered, the solvent was evaporated under reduced pressure, and dried at 35°C for 4h to obtain compound VI. Compound VI does not need to be purified and can be directly used in the next step.

[0082] Add compound VI obtained in the previous step to a reaction flask, add anhydrous ethanol (30 mL), add 1.0 mol / L NaOH / EtOH (11 mL) dropwise, after the addition is complete, heat to 45-50 ° C, react for 6-12 hours, and monitor the reaction by TLC (petroleum ether: ethyl acetate = 5: 1-2: 1, v / v). After the reaction is completed, evaporate the solvent under reduced pressure to obtain a yellow or white solid residue. Add water (10 mL), and slowly add 1 mol / L HCl under an ice-water bath to adjust the pH to 1-2. Add 120 mL of water, extract with ethyl acetate (40 mL × 3), combine the organic phases, wash with saturated NaCl (40 mL), add anhydrous MgSO4 to dry, filter, evaporate the solvent under reduced pressure, use methanol / dichloromethane as the eluent, and column chromatography to obtain compound Z1.

[0083] The physicochemical and spectral data of compound Z1 are as follows:

[0084]

[0085] White solid, yield 64%, mp 218.5-220.7℃; 1 H NMR (400MHz, DMSO-d6) δ12.98(s,1H),10.15(s,1H),8.24(s,1H),8.11–7.38(m,6H),7.28–7.01(m,3H),6.84( d,J=8.6Hz,2H),3.98(t,J=6.3Hz,2H),2.60–2.46(m,4H),2.01(dd,J=13.7,6.9Hz,2H),1.13(t,J=7.5Hz,3H); 13C NMR (151MHz, DMSO-d6) δ171.15,167.65,167.47,157.02,139.66,138.04,136.07,132.09,131.68,129.29,129. 23,129.02,126.49,125.11,123.73,122.56,121.76,114.76,114.36,67.08,31.82,27.74,25.13,16.36; ESI-MS m / z:445.33[MH] - .

[0086] 2. The preparation methods, physicochemical and spectral data of compounds Z2 to Z53 are as follows:

[0087] 3-(2-(4-(4-acetylphenoxy)butyrylamino)benzamido)benzoic acid (Z2)

[0088]

[0089] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-acetylphenoxy)butyric acid (6c), and the other conditions were the same. Compound Z2: white solid, yield 66%, mp 192.9-193.9℃; 1 H NMR(600MHz,DMSO-d6)δ13.00(s,1H),10.56(s,1H),10.43(s,1H),8.40(s,1H),8.07(d,J =8.1Hz,1H),7.93(d,J=7.8Hz,1H),7.87(d,J=8.7Hz,2H),7.77(d,J=7.7Hz,1H),7.69(d, J=7.7Hz,1H),7.53(t,J=7.7Hz,1H),7.46(t,J=7.9Hz,1H),7.24(t,J=7.5Hz,1H),6.98(d ,J=8.7Hz,2H),4.08(t,J=6.4Hz,2H),2.58–2.50(m,2H),2.49(s,3H),2.05–2.02(m,2H); 13C NMR(151MHz,DMSO-d6)δ196.69,171.04,167.66,167.46,162.85,139.66,137.98,132.08,131.70,130.89,130.2 6,129.29,129.23,125.46,125.11,125.05,123.79,122.63,121.73,114.68,67.55,33.50,26.83,24.91; ESI-MS m / z:461.15[M+H] + .

[0090] 3-(2-(5-(4-acetylphenoxy)pentanoylamino)benzamido)benzoic acid (Z3)

[0091]

[0092] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-acetylphenoxy)pentanoic acid (6d), and the other conditions were the same. Compound Z3: white solid, yield 74%, mp 176.6-179.4℃; 1 H NMR (600MHz, DMSO-d6) δ10.57(s,1H),10.39(s,1H),8.41(s,1H),8.09(d,J=8.2Hz,1H),7.97–7.86(m,3H),7.79–7.75(m,1H),7.70(d,J=7.8H z,1H),7.55–7.45(m,2H),7.27–7.22(m,1H),7.05–6.96(m,2H),4.03( t,J=6.0Hz,2H),2.50(s,3H),2.41(t,J=7.0Hz,2H),1.79–1.71(m,4H); 13 C NMR(151MHz,DMSO-d6)δ196.73,171.46,167.66,167.51,162.96,139.63,138.05,132.10,131.68,130.90,130.20,1 29.31,129.20,125.31,125.15,125.12,123.74,122.57,121.78,114.67,67.96,36.71,28.33,26.84,22.05; ESI-MS m / z:473.26[MH] - .

[0093] 3-(2-(4-(tert-butyl)phenoxy)butyrylamino)benzamido)benzoic acid (Z4)

[0094]

[0095] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-(tert-butyl)phenoxy)butyric acid (6e), and the other conditions were the same. Compound Z4: white solid, yield 78%, mp 122.1-123.5℃; 1 H NMR(600MHz,DMSO-d6)δ10.56(s,1H),10.42(s,1H),8.40(s,1H),8.08(d,J=8 .2Hz,1H),7.95(d,J=8.0Hz,1H),7.77(d,J=6.9Hz,1H),7.70(d,J=7.7Hz,1H) ,7.56–7.50(m,1H),7.46(t,J=7.9Hz,1H),7.29–7.19(m,3H),6.86–6.77(m,2 H),3.94(t,J=6.4Hz,2H),2.53–2.47(m,2H),2.06–1.96(m,2H),1.22(s,9H); 13 C NMR (151MHz, DMSO-d6) δ196.72,171.44,167.66,162.96,139.65,138.04,134.51,132.09,131.66,130.94,130.91,130.18,129. 31,125.13,123.73,121.75,120.25,114.71,114.66,67.95,37.44,36.69,36.39,28.33,26.86,22.04; ESI-MSm / z:475.14[M+H] + .

[0096] 3-(2-(5-(4-(tert-butyl)phenoxy)pentanoylamino)benzamido)benzoic acid (Z5)

[0097]

[0098] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-(tert-butyl)phenoxy)pentanoic acid (6f), and the other conditions were the same. Compound Z5: white solid, yield 77%, mp 188.4-191.1℃; 1H NMR (600MHz, DMSO-d6) δ10.57(s,1H),10.39(s,1H),8.41(s,1H),8.11(d,J=8.2Hz,1H),7.99–7.91(m,1H),7.80–7.67(m,2H),7. 56–7.49(m,1H),7.47(t,J=7.9Hz,1H),7.26–7.21(m,3H),6.82–6.76(m,2H),3.89(s,2H),2.40(s,2H),1.72(s,4H),1.23(s,9H); 13 C NMR(151MHz,DMSO-d6)δ171.51,167.65,167.52,156.80,142.94,139.62,138.10,132.11,131.69,129.31, 129.21,126.44,125.14,123.70,122.52,121.80,114.31,67.40,36.79,34.17,31.82,28.53,22.19; ESI-MS m / z:489.41[M+H] + .

[0099] 3-(2-(5-(4-ethylphenoxy)pentanoylamino)benzamido)benzoic acid (Z6)

[0100]

[0101] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-ethylphenoxy)pentanoic acid (6b), and the other conditions were the same. Compound Z6: white solid, yield 57%, mp 205-206.5℃; 1 H NMR(600MHz,DMSO-d6)δ12.97(s,1H),10.11(s,1H),8.48–8.04(m,2H),7.99–7.75(m,2H),7.66(dd,J=9.5,7.7Hz,2H),7.55–7.30(m,2 H),7.08(t,J=9.0Hz,3H),6.81(dd,J=9.5,8.4Hz,3H),3.95-3.90(m,2H),2.52-2.31(m,4H),1.72-1.68(m,4H),1.13(t,J=7.6Hz,3H); 13C NMR (151MHz, DMSO) δ171.81,171.49,167.67,157.14,139.96,139.64,138.12,136.04,132.11,131.72,129.39,12 9.22,129.06,125.15,124.26,123.59,121.79,120.27,114.74,67.49,36.47,28.74,27.76,22.22,16.41; ESI-MS m / z:461.38[M+H].

[0102] 3-(2-(4-(4-propionylphenoxy)butyrylamino)benzamido)benzoic acid (Z7)

[0103]

[0104] According to the preparation method of compound Z1, compound 6a was replaced with 4-(4-propionylphenoxy)butyric acid (6 g), and the other conditions were the same. Compound Z7: white solid, yield 72%, mp 172.3-175.8°C; 1 H NMR(600MHz,DMSO-d6)δ13.03(s,1H),10.57(s,1H),10.44(s,1H),8.41(s,1H),8.14–8.00(m,1H),7.99–7.67(m,5H),7.65–7.39(m,2H ),7.38–7.12(m,1H),7.05–6.82(m,2H),4.16–4.00(m,2H),3.00–2.91(m,2H),2.59–2.49(m,2H),2.15–2.00(m,2H),1.11–1.01(m,3H); 13 C NMR(151MHz,DMSO-d6)δ196.72,171.44,167.66,167.50,162.96,139.65,138.04,134.51,132.09,131.66,130.94,1 30.91,130.18,129.31,125.13,123.73,121.75,120.25,114.71,114.66,67.95,36.69,28.33,26.86,22.04; ESI-MS m / z:473.25[MH] - .

[0105] 3-(2-(5-(4-propionylphenoxy)pentanoylamino)benzamido)benzoic acid (Z8)

[0106]

[0107] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-propionylphenoxy)pentanoic acid (6h), and the other conditions were the same. Compound Z8: white solid, yield 75%, mp 253.6-255.5℃; 1 H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.57(s,1H),10.10(s,1H),8.41(s,1 H),8.24(s,1H),8.10(d,J=8.2Hz,1H),8.00–7.85(m,3H),7.85–7.07(m,4H), 7.03(dd,J=9.1,2.3Hz,1H),6.98(d,J=8.9Hz,1H),4.06(m,2H),3.00–2.81( m,2H),2.41(dd,J=9.0,6.9Hz,2H),1.85–1.61(m,4H),1.07(t,J=7.2Hz,3H); 13 C NMR(151MHz,DMSO-d6)δ199.25,171.76,171.44,167.65,162.81,139.95,139.63,138.09,132.10,131.69,130.54,130 .50,129.92,129.39,125.14,123.60,121.78,120.28,114.73,114.68,67.99,36.40,31.23,28.53,22.10,8.78; ESI-MS m / z:489.36[M+H] + .

[0108] 3-(2-(4-(4-(Benzyloxy)phenoxy)butyrylamino)benzylamino)benzoic acid (Z9)

[0109]

[0110] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-(benzyloxy)phenoxy)butyric acid (6i), and the other conditions were the same. Compound Z9: white solid, yield 62%, mp 196.3-199.5℃; 1H NMR(600MHz,DMSO-d6)δ12.99(s,1H),10.54(s,1H),10.39(s,1H),8.38(s,1 H),8.06(d,J=8.2Hz,1H),7.92(d,J=7.9Hz,1H),7.74(d,J=7.3Hz,1H),7.67 (d,J=7.7Hz,1H),7.52–7.25(m,7H),7.21(t,J=7.4Hz,1H),6.85–6.75(m,4H ),4.97(s,2H),3.88(t,J=6.4Hz,2H),2.53–2.35(m,2H),2.03–1.64(m,2H); 13 C NMR(151MHz,DMSO-d6)δ171.16,167.67,167.49,153.16,152.79,139.67,138.05,137.83,132.10,131.70,129.31,129.24,1 28.84,128.17,128.07,125.32,125.13,125.08,123.73,122.56,121.75,116.08,115.77,70.09,67.57,33.69,25.17; ESI-MS m / z:525.32[M+H] + .

[0111] 3-(2-(5-(4-(Benzyloxy)phenoxy)pentanoylamino)benzamido)benzoic acid (Z10)

[0112]

[0113] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-(benzyloxy)phenoxy)pentanoic acid (6J), and the other conditions were the same. Compound Z10: white solid, yield 66%, mp 163.5-165.2℃; 1 H NMR (600MHz, DMSO-d6) δ12.99(s,1H),10.55(s,1H),10.36(s,1H),8.38(s,1H),8.08(d,J=8.1Hz,1H),7.92(d,J=7.7Hz,1H),7. 83–7.55(m,2H),7.52–7.18(m,7H),6.90–6.70(m,4H),4.98(d,J=6.1Hz,2H),3.91–3.72(m,2H),2.36(s,2H),1.79–1.54(m,4H); 13C NMR (151MHz, DMSO-d6) δ171.50,167.68,167.52,153.26,152.72,139.64,138.12,137.85,132.11,131.73,129.31,129.22,128. 85,128.17,128.07,125.15,125.11,123.68,122.52,121.79,116.15,116.11,115.70,70.10,67.91,36.81,28.60,22.18; ESI-MS m / z:539.40[M+H] + .

[0114] 3-(2-(4-([1,1'-biphenyl]-2-oxy)butyrylamino)benzylamino)benzoic acid (Z11)

[0115]

[0116] According to the preparation method of compound Z1, compound 6a was replaced by 4-([1,1'-biphenyl]-2-oxy)butyric acid (6k), and the other conditions were the same. Compound Z11: white solid, yield 61%, mp 208.1-210.2℃; 1 H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.59(s,1H),10.12(s,1H),8.43(s,1H),8.24–8.07(m,1H),8.02–7.90(m,1H),7.84–7.59(m ,2H),7.56–7.36(m,5H),7.34–7.19(m,3H),7.15–6.94(m,2H),4.08–3.98(m,2H),2.45(dd,J=12.7,7.2Hz,2H),1.99–1.91(m,2H); 13 C NMR (151MHz, DMSO-d6) δ171.04,167.66,167.56,155.78,139.62,138.60,138.18,132.19,131.71,130.87,130.42,129.69,129.3 0,129.25,128.38,127.22,126.49,125.18,125.13,124.90,123.67,122.37,121.83,121.29,113.20,67.46,33.62,25.05; ESI-MS m / z:493.16[MH] - .

[0117] 3-(2-(5-([1,1'-biphenyl]-2-oxy)pentanoylamino)benzamido)benzoic acid (Z12)

[0118]

[0119] According to the preparation method of compound Z1, compound 6a was replaced by 5-([1,1'-biphenyl]-2-oxy)pentanoic acid (6l), and the other conditions were the same. Compound Z11: white solid, yield 76%, mp 137.2-139.3℃; 1 H NMR(600MHz,DMSO-d6)δ13.02(s,1H),10.57(s,1H),10.36(s,1H),8.41(s,1H),8.25–8.05(m,1H),7.99–7.76(m,2H),7.71–7.59(m,1H) ),7.56–7.46(m,4H),7.37-7.28(m,2H)7.29-7.22(m,4H),7.13–6.98(m,2H),4.03–3.88(m,2H),2.38–2.30(m,2H),1.77–1.57(m,4H); 13 C NMR(151MHz,DMSO-d6)δ171.46,167.65,167.51,155.90,139.63,138.62,138.14,132.11,131.70,130.85,130.38,129.69,129.29,1 29.24,129.21,128.32,127.21,127.17,125.14,123.67,122.51,121.80,121.25,121.21,113.19,67.81,36.63,28.46,22.05; ESI-MS m / z:509.19[M+H] + .

[0120] 3-(2-(4-(2,4-di-tert-butylphenoxy)butyrylamino)benzamido)benzoic acid (Z13)

[0121]

[0122] According to the preparation method of compound Z1, compound 6a was replaced by 4-(2,4-di-tert-butylphenoxy)butyric acid (6m), and the other conditions were the same. Compound Z13: white solid, yield 52%, mp 125.5-127.2℃; 1H NMR(600MHz,DMSO-d6)δ12.97(s,1H),10.56(s,1H),10.16(s,1H),8.40(s,1H),8.24(s,1H),8.0 9(d,J=8.2Hz,1H),7.95(d,J=8.1Hz,1H),7.78(dd,J=7.8,1.1Hz,1H),7.72–7.67(m,1H),7.47–7. 41(m,1H),7.20(d,J=2.5Hz,1H),7.08(dd,J=8.5,2.4Hz,1H),6.87(d,J=8.6Hz,1H),6.78(d,J=8 .6Hz,1H),3.97(t,J=6.4Hz,2H),2.58–2.54(m,2H),2.11–2.05(m,2H),1.32(s,9H),1.23(s,9H); 13 C NMR(151MHz,DMSO-d6)δ171.05,167.64,167.48,155.42,142.18,139.93,139.64,138.02,136.60,132.11,131.70,129.26,125.12,125.0 6,123.91,123.85,123.38,123.32,121.76,120.33,112.25,67.15,35.06,35.00,34.32,33.96,33.53,31.90,30.25,25.45,25.33; ESI-MS m / z:529.64[MH] - .

[0123] 3-(2-(5-(2,4-di-tert-butylphenoxy)pentanoylamino)benzamido)benzoic acid (Z14)

[0124]

[0125] According to the preparation method of compound Z1, compound 6a was replaced by 5-(2,4-di-tert-butylphenoxy)pentanoic acid (6n), and the other conditions were the same. Compound Z14: white solid, yield 55%, mp 98.3-101.2℃; 1H NMR(600MHz,DMSO-d6)δ13.06(s,1H),8.41–8.38(m,1H),8.13–7.93(m,2H),7.88–7.64(m,3H),7.55–7.43(m,1H),7.25– 7.09(m,3H),6.80–6.58(m,1H),4.06–3.85(m,2H),2.43–2.35(m,2H),1.88–1.70(m,4H),1.32-1.30(m,9H),1.24(s,9H); 13 C NMR(151MHz,DMSO-d6)δ168.45,167.01,161.99,156.67,155.52,150.38 ,147.69,142.10,138.11,136.57,130.39,129.21,127.41,126.98,125. 12,124.62,123.84,123.31,116.90,115.13,112.18,67.65,36.75,35.2 7,35.02,34.99,34.33,31.91,30.23,30.19,28.85,23.31,22.47; ESI-MS m / z:543.56[MH] - .

[0126] 3-(2-(4-(Naphthalene-2-yl)butyrylamino)benzamido)benzoic acid (Z15)

[0127]

[0128] According to the preparation method of compound Z1, compound 6a was replaced by 4-(naphthalene-2-oxy)butyric acid (6o), and the other conditions were the same. Compound Z15: white solid, yield 66%, mp 163.2-165.7℃; 1 H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.17(s,1H),8.52–8.40(m,1H),8.27(s,1H),8.14–7.93(m,1H),7.87–7.67(m,5H),7.65 –7.52(m,1H),7.49–7.39(m,2H),7.36–7.22(m,3H),7.18–7.12(m,1H),4.18–4.11(m,2H),2.66–2.55(m,2H),2.17–2.08(m,2H); 13C NMR(151MHz,DMSO-d6)δ171.47,170.02,167.67,156.93,141.29,139.96,134.78,134.36,131.72,131.54,129.73,129.40,12 8.92,127.96,127.13,126.82,125.13,124.30,123.99,123.63,123.01,120.31,119.17,107.20,67.45,33.35,25.05; ESI-MS m / z:469.29[M+H] + .

[0129] 3-(2-(5-(Naphthalene-2-yl)pentanoylamino)benzamido)benzoic acid (Z16)

[0130]

[0131] According to the preparation method of compound Z1, compound 6a was replaced by 4-(naphthalene-2-oxy)pentanoic acid (6p), and the other conditions were the same. Compound Z16: white solid, yield 65%, mp 165.5-167.5℃; 1 H NMR(600MHz,DMSO-d6)δ13.01(s,1H),10.09(s,1H),8.22(s,1H),8.11–7.96(m,1H),7.93–7.55(m,6H ),7.51–7.24(m,4H),7.23–6.97(m,2H),4.11–3.98(m,2H),2.39(t,J=6.6Hz,2H),1.83–1.71(m,4H); 13 CNMR(151MHz,DMSO-d6)δ171.82,167.68,157.01,139.98,134.78,131.73,129.71,129.67,129.40,128.89,128.86,127.96,127. 11,126.80,125.14,124.28,123.94,123.61,121.81,120.29,119.23,119.17,107.10,107.05,67.66,36.47,28.66,22.25; ESI-MS m / z:481.41[MH] - .

[0132] 4-(2-(5-(4-ethylphenoxy)pentanoylamino)benzamido)benzoic acid (Z17)

[0133]

[0134] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-acetylphenoxy)butyric acid (6b), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z17: white solid, yield 77%, mp 171.9-172.5℃; 1 H NMR(600MHz,DMSO-d6)δ12.73(s,1H),10.23(s,1H),8.05–7.85(m,4H),7.75–7.56(m,2H),7.55–7.22(m,1 H),7.16–6.76(m,5H),3.95(s,2H),2.57–2.50(m,2H),2.46–2.36(m,2H),1.75(s,4H),1.18–1.11(m,3H); 13 C NMR(151MHz,DMSO-d6)δ172.11,171.50,167.41,157.13,143.80,137.81,136.04,132.07,130.84,130.66,129.26,129.07, 129.03,126.06,125.32,123.84,122.99,120.04,118.71,114.73,114.69,67.47,36.56,28.73,27.76,22.15,16.43; ESI-MS m / z:461.18[M+H] + .

[0135] 4-(2-(4-(4-acetylphenoxy)butyrylamino)benzamido)benzoic acid (Z18)

[0136]

[0137] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-acetylphenoxy)butyric acid (6c), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z18: white solid, yield 70%, mp 152.3-154.5℃; 1H NMR(600MHz,DMSO-d6)δ11.21(s,1H),8.49(d,J=8.2Hz,1H),8.03–7.84(m,5H),7.71(d,J=8.7Hz,1H),7.64 –7.54(m,1H),7.20–6.98(m,4H),4.14(t,J=6.4Hz,2H),2.61–2.55(m,2H),2.51(s,3H),2.16–2.03(m,2H); 13 C NMR(151MHz,DMSO-d6)δ196.73,171.65,171.15,169.99,167.40,162.84,143.75,141.23,134.39,131.52,1 30.93,130.84,130.33,125.39,123.04,120.49,118.75,117.22,114.73,67.56,34.21,26.85,24.79; ESI-MS m / z:461.30[M+H] + .

[0138] 4-(2-(5-(4-acetylphenoxy)pentanoylamino)benzamido)benzoic acid (Z19)

[0139]

[0140] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-acetylphenoxy)pentanoic acid (6d), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z19: white solid, yield 79%, mp 180.3-182.6℃; 1 H NMR(600MHz,DMSO-d6)δ12.76(s,1H),10.24(s,1H),8.05–7.96(m,1H),7.96–7.82(m,5H),7.73(t,J=9.9Hz,2H) ,7.63–7.48(m,1H),7.29–6.92(m,3H),4.10(t,J=5.8Hz,2H),2.51(s,3H),2.47–2.38(m,2H),1.82–1.71(m,4H); 13C NMR (151MHz, DMSO-d6) δ196.72,172.06,167.58,167.42,162.96,143.79,137.76,132.06,131.54,130.94,130.85,130. 66,130.23,129.25,126.06,125.33,123.88,120.02,118.72,114.70,114.65,68.00,36.49,28.52,26.85,22.03; ESI-MS m / z:475.11[M+H] + .

[0141] 4-(2-(4-(tert-butyl)phenoxy)butyrylamino)benzamido)benzoic acid (Z20)

[0142]

[0143] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-(tert-butyl)phenoxy)butyric acid (6e), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z20: white solid, yield 73%, mp 209.6-210.15℃; 1 H NMR(600MHz,DMSO-d6)δ12.73(s,1H),10.64(s,1H),10.28(s,1H),8.00–7.80(m,4H),7.70(s,1 H),7.50(s,1H),7.19(s,3H),6.76(s,2H),3.91(s,2H),2.47(s,2H),1.96(s,2H),1.20(s,9H); 13 C NMR(151MHz,DMSO-d6)δ171.16,167.55,167.43,156.70,143.69,143.04,137.71,132.04,130.84,130.67,129.27,1 26.51,126.44,126.24,126.04,123.93,122.95,119.98,118.74,114.35,67.04,34.17,33.54,31.80,25.14; ESI-MS m / z:475.43[M+H] + ; 473.47 [MH] - .

[0144] 4-(2-(5-(4-(tert-butyl)phenoxy)pentanoylamino)benzamido)benzoic acid (Z21)

[0145]

[0146] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-(tert-butyl)phenoxy)pentanoic acid (6f), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z21: white solid, yield 75%, mp 137.7-139.3℃; 1 H NMR(600MHz,DMSO-d6)δ12.72(s,1H),10.23(s,1H),8.06–7.85(m,4H),7.76–7.20(m,5H) ,6.86–6.56(m,3H),3.96(s,2H),2.48–2.38(m,2H),1.78–1.70(m,4H),1.31–1.20(m,9H); 13 C NMR (151MHz, DMSO-d6) δ172.11,168.59,167.42,156.81,150.42,143.81,142.99,132.92,130.85,130.67,130.60,129. 35,126.49,125.32,120.05,119.98,118.71,116.90,115.14,114.34,67.42,36.55,34.19,31.82,28.72,22.16; ESI-MS m / z:489.13[M+H] + .

[0147] 4-(2-(4-(4-ethylphenoxy)butyrylamino)benzamido)benzoic acid (Z22)

[0148]

[0149] According to the preparation method of compound Z1, compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z22: white solid, yield 70%, mp 184.4-186.7℃; 1H NMR(600MHz,DMSO-d6)δ12.73(s,1H),10.65(s,1H),10.26(s,1H),8.13–7.98 (m,1H),7.94–7.68(m,5H),7.66–7.51(m,1H),7.29–7.21(m,1H),7.10(d,J=8. 6Hz,1H),7.05(d,J=8.6Hz,1H),6.85–6.76(m,2H),3.98(t,J=6.3Hz,1H),3.94 (t,J=6.5Hz,1H),2.54–2.47(m,4H),2.04–1.97(m,2H),1.13(q,J=7.6Hz,3H); 13 C NMR (151MHz, DMSO-d6) δ171.78,171.16,167.42,157.02,143.67,137.73,136.07,132.05,130.83,130.65,129. 09,129.01,126.05,125.36,123.91,120.00,118.74,114.78,114.75,67.09,33.45,27.74,25.14,16.34; ESI-MS m / z:447.37[M+H] + , 445.48[MH] - .

[0150] 4-(2-(4-(4-propionylphenoxy)butyrylamino)benzamido)benzoic acid (Z23)

[0151]

[0152] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-propionylphenoxy)butyric acid (6g), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z23: white solid, yield 66%, mp 139.1-140.5℃; 1 H NMR(600MHz,DMSO-d6)δ12.56(s,1H),10.67(s,1H),10.33(s,1H),8.02–7.70(m,7H),7.68–7.52(m,1H),7.30–7.12(m,1H),7.07– 6.75(m,3H),4.09–4.03(m,2H),3.00–2.93(m,2H),2.62–2.54(m,1H),2.40(t,J=7.3Hz,1H),2.07–1.95(m,2H),1.09–1.04(m,3H);13 C NMR (151MHz, DMSO-d6) δ199.25,174.52,171.05,167.53,167.43,162.69,143.69,137.65,132.04,130.65,130.55,130.47,129. 97,129.91,129.27,126.00,123.96,122.99,119.94,114.71,114.65,67.50,31.24,30.47,24.56,8.76; ESI-MSm / z:473.20[MH] - .

[0153] 4-(2-(5-(4-propionylphenoxy)pentanoylamino)benzamido)benzoic acid (Z24)

[0154]

[0155] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-propionylphenoxy)pentanoic acid (6h), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z24: white solid, yield 70%, mp 187.6-190.4℃; 1 H NMR(600MHz,DMSO-d6)δ12.71(s,1H),10.66(s,1H),10.22(s,1H),8.02(d,J=8.2Hz ,1H),7.96–7.84(m,5H),7.76–7.69(m,2H),7.56–7.50(m,1H),7.27–7.21(m,1H),7. 03(d,J=8.9Hz,1H),6.98(d,J=8.9Hz,1H),4.09(t,J=5.9Hz,1H),4.02(t,J=6.0Hz, 1H),2.98–2.94(m,2H),2.43–2.40(m,2H),1.81–1.72(m,4H),1.07(t,J=7.2Hz,3H); 13C NMR(151MHz,DMSO-d6)δ199.26,172.06,171.46,167.58,167.40,162.82,143.78,137.80,132.07,130.83,130.65,130.54,130 .49,129.92,129.24,126.09,125.35,123.87,120.06,118.73,114.73,114.67,67.98,36.49,31.24,28.52,22.03,8.78; ESI-MS m / z:489.37[M+H] + .

[0156] 4-(2-(4-(Benzyloxy)phenoxy)butyrylamino)benzamido)benzoic acid (Z25)

[0157]

[0158] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-(benzyloxy)phenoxy)butyric acid (6i), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z25: white solid, yield 78%, mp 191.5-192.5℃; 1 H NMR(600MHz, DMSO-d6)δ12.65(s,1H),10.32(d,J=4.8Hz,1H),8.07–7.87(m,4H),7.81–7.55(m,2H),7.51–7.12(m ,6H),6.99–6.79(m,5H),6.68–6.57(m,1H),5.07(s,2H),4.03–3.90(m,2H),2.65–2.49(m,2H),2.14–1.88(m,2H); 13 CNMR(151MHz,DMSO-d6)δ171.79,171.17,167.44,153.17,152.80,143.79,143.67,137.83,137.76,132.07,130.84,130.67,129.27,1 28.84,128.17,128.07,126.06,125.36,123.89,120.02,118.75,116.18,116.09,115.80,115.78,70.11,67.59,33.46,25.18; ESI-MS m / z:525.39[M+H] + .

[0159] 4-(2-(5-(4-(Benzyloxy)phenoxy)pentanoylamino)benzamido)benzoic acid (Z26)

[0160]

[0161] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-(benzyloxy)phenoxy)pentanoic acid (6j), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z26: white solid, yield 75%, mp 178.3-180.1℃; 1 H NMR (600MHz, DMSO-d6) δ12.69(s,1H),10.23(s,1H),7.80(dd,J=10.2,7.9Hz,5H),7.52–7.09(m,7H),6. 89(dd,J=8.8,7.8Hz,5H),5.03(s,2H),3.92(s,2H),3.37(s,2H),2.46–2.30(m,2H),1.74–1.63(m,4H); 13 C NMR(151MHz,DMSO-d6)δ172.12,171.58,170.06,167.42,153.27,152.76,143.81,141.34,137.85,134.36,131.55,130.84,1 28.84,128.17,128.07,125.34,122.94,120.38,120.07,118.73,116.15,115.75,70.10,67.97,36.57,28.77,22.15; ESI-MS m / z:539.18[M+H] + .

[0162] 4-(2-(4-([1,1'-biphenyl]-2-oxy)butyrylamino)benzamido)benzoic acid (Z27)

[0163]

[0164] According to the preparation method of compound Z1, compound 6a was replaced by 4-([1,1'-biphenyl]-2-oxy)butyric acid (6k), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z27: white solid, yield 70%, mp 182.2-184.2℃; 1H NMR(600MHz,DMSO-d6)δ12.71(s,1H),10.23(s,1H),8.04–7.86(m,4H),7.76–7.69(m,2H),7.55–7.36( m,5H),7.34–7.18(m,4H),7.13–6.98(m,2H),4.08–3.98(m,2H),2.49–2.41(m,2H),2.00–1.89(m,2H); 13 C NMR(151MHz,DMSO-d6)δ171.68,171.07,167.41,155.83,143.75,143.62,138.63,137.85,132.14,130.96,130.83,130.64,130.38,129.7 1,129.31,129.24,128.43,128.38,127.27,126.11,125.37,123.86,121.37,121.29,120.06,118.76,113.31,67.76,33.29,24.93; ESI-MS m / z:495.03[M+H] + .

[0165] 4-(2-(5-([1,1'-biphenyl]-2-oxy)pentanoylamino)benzamido)benzoic acid (Z28)

[0166]

[0167] According to the preparation method of compound Z1, compound 6a was replaced by 5-([1,1'-biphenyl]-2-oxy)pentanoic acid (6l), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z28: white solid, yield 66%, mp 135.4-137.2℃; 1 H NMR(600MHz,DMSO-d6)δ12.76(s,1H),10.25(s,1H),8.07–7.90(m,2H),7.81–7.67(m,2H),7. 63–7.49(m,3H),7.45–7.25(m,6H),7.19–7.03(m,3H),4.05(s,2H),2.42(s,2H),1.75(s,4H); 13C NMR(151MHz,DMSO-d6)δ172.09,171.54,170.10,167.43,155.91,143.81,141.38,138.64,134.25,131.56,130.90,130.85,130.60,1 30.39,129.71,129.27,128.34,127.22,125.34,122.88,121.25,120.29,120.06,118.72,113.20,67.79,36.40,28.63,22.08; ESI-MS m / z:509.20[M+H] + .

[0168] 4-(2-(4-(2,4-di-tert-butylphenoxy)butyrylamino)benzamido)benzoic acid (Z29)

[0169]

[0170] According to the preparation method of compound Z1, compound 6a was replaced by 4-(2,4-di-tert-butylphenoxy)butyric acid (6m), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z29: white solid, yield 62%, mp 194.5-196.7℃; 1 H NMR(600MHz,DMSO-d6)δ12.77(s,1H),10.66(s,1H),10.32(s,1H),8.01–7.84(m,3H),7.76–7.70(m,1H),7.61–7.50(m,1H) ,7.28–7.05(m,3H),6.89–6.74(m,1H),4.06–3.90(m,2H),2.66–2.52(m,2H),2.16–1.96(m,2H),1.34(s,9H),1.24(s,9H); 13 C NMR(151MHz,DMSO-d6)δ171.07,167.54,167.41,155.43,143.66,142.18,137.66,136.56,132.04,130.83,130.63,129.24,12 6.05,123.99,123.85,123.33,119.98,118.78,112.28,67.18,35.00,34.33,33.79,33.63,31.90,30.31,30.25,25.48; ESI-MS m / z:531.37[M+H] + .

[0171] 4-(2-(5-(2,4-di-tert-butylphenoxy)pentanoylamino)benzamido)benzoic acid (Z30)

[0172]

[0173] According to the preparation method of compound Z1, compound 6a was replaced by 5-(2,4-di-tert-butylphenoxy)pentanoic acid (6n), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z30: white solid, yield 65%, mp 193.7-195.5℃; 1 H NMR (600MHz, DMSO-d6) δ12.76(s,1H),10.68(s,1H),10.27(d,J=14.3Hz,1H),8.10–7.79(m,3H),7.72(d,J=7.8Hz,1H),7.55(d,J=9.2Hz,1H ),7.38–7.03(m,3H),6.82(dd,J=9.7,7.6Hz,1H),3.94(d,J=8.9Hz,2H),2.45-2.42(m,2H),1.81(d,J=9.9Hz,4H),1.33(s,9H),1.24(s,9H); 13 C NMR(151MHz,DMSO-d6)δ172.05,171.45,167.42,155.52,143.63,142.08 ,137.83,136.54,132.06,130.83,130.64,129.24,126.12,125.37,123. 84,123.35,123.30,120.06,118.73,112.14,67.36,36.59,36.45,35.05 ,35.00,34.32,31.91,30.24,30.20,29.00,28.81,22.45,22.41; ESI-MS m / z:543.62[MH] - .

[0174] 4-(2-(4-(Naphthalene-2-yl)butyrylamino)benzamido)benzoic acid (Z31)

[0175]

[0176] According to the preparation method of compound Z1, compound 6a was replaced by 4-(naphthalene-2-oxy)butyric acid (6o), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z31: white solid, yield 61%, mp 218.7-221.3℃; 1 H NMR(600MHz,DMSO-d6)δ12.75(s,1H),10.31(s,1H),8.02–7.87(m,3H),7.85–7.71(m,6H),7.66–7.41(m,2H) ,7.37–7.28(m,3H),7.15(dd,J=8.9,2.3Hz,1H),4.15(t,J=6.2Hz,2H),2.65–2.56(m,2H),2.18–2.10(m,2H); 13 C NMR(151MHz,DMSO-d6)δ171.77,171.24,170.04,167.42,156.92,143.80,141.30,134.77,134.35,131.55,130.86,129.73,1 28.92,127.96,127.13,126.83,125.37,123.99,123.00,120.43,119.17,118.75,107.16,67.42,34.40,33.47,24.96; ESI-MS m / z:491.11[M+Na] + .

[0177] 4-(2-(5-(Naphthalene-2-yl)pentanoylamino)benzamido)benzoic acid (Z32)

[0178]

[0179] According to the preparation method of compound Z1, compound 6a was replaced by 4-(naphthalene-2-oxy)pentanoic acid (6p), and compound 5a was replaced by ethyl 4-(2-aminobenzamido)benzoate (5b), and the other conditions were the same. Compound Z32: white solid, yield 60%, mp 159.3-160.8℃; 1 H NMR (600MHz, DMSO-d6) δ12.73(s,1H),10.26(s,1H),8.11–7.66(m,9H),7.57(s,1H),7.45(t,J=7.2Hz,1H) ,7.38–7.22(m,3H),7.19–7.09(m,1H),4.12(t,J=5.6Hz,2H),2.46(t,J=6.7Hz,2H),1.84(d,J=5.7Hz,4H);13 C NMR(151MHz,DMSO-d6)δ172.12,167.42,156.99,143.81,134.78,130.86,130.68,129.72,128.88,127.96,127.11,1 26.81,125.33,123.95,122.96,120.40,120.05,119.23,118.73,107.08,67.64,60.23,36.56,28.65,22.18; ESI-MS m / z:481.17[MH] - .

[0180] 2-(2-(5-(4-ethylphenoxy)pentanoylamino)benzamido)benzoic acid (Z33)

[0181]

[0182] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-ethylphenoxy)pentanoic acid (6b), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z33: white solid, yield 82%, mp 171.8-174.6℃; 1 H NMR(600MHz,DMSO-d6)δ13.72(s,1H),11.88(s,1H),10.54(s,1H),8.61(d,J=8.3Hz,1H),8.09 (d,J=8.2Hz,1H),8.04(d,J=7.4Hz,1H),7.79(d,J=7.6Hz,1H),7.65(t,J=7.6Hz,1H),7.56(t,J =7.7Hz,1H),7.27(t,J=7.5Hz,1H),7.22(t,J=7.6Hz,1H),7.07(d,J=8.3Hz,2H),6.80(d,J=8.4 Hz,2H),3.89(t,2H),2.54–2.51(m,2H),2.39(t,2H),1.75-1.67(m,4H),1.13(t,J=7.6Hz,3H); 13C NMR(151MHz,DMSO-d6)δ171.53,170.15,166.93,157.13,141.11,138.17,136.00,134.54,132.54,131.63,129.02, 128.40,125.31,124.19,123.71,122.91,120.98,117.74,114.72,67.46,36.81,28.59,27.75,22.09,16.39; ESI-MS m / z:459.30[MH] - .

[0183] 2-(2-(4-(4-acetylphenoxy)butyrylamino)benzamido)benzoic acid (Z34)

[0184]

[0185] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-acetylphenoxy)butyric acid (6c), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z34: white solid, yield 55%, mp 83.7-86.9℃; 1 H NMR (600MHz, DMSO-d6) δ13.27(s,1H),10.90(s,1H),8.58(d,J=8.1Hz,1H),8.19(s,1H),8.04(d,J=7.5Hz,1H),7.87-7.84(m,3H),7.61– 7.44(m,2H),7.24(t,J=7.5Hz,1H),7.14(s,1H),6.99(s,2H),4.09(t,J=6.1Hz,2H),2.57–2.51(m,2H),2.50(s,3H),2.13–1.93(m,2H); 13 C NMR(151MHz,DMSO-d6)δ195.66,170.03,165.90,161.82,140.07,137.63,131.45,130.66,129.8 7,129.25,127.39,122.98,122.23,121.44,119.19,113.68,66.56,32.72,25.83,23.84; ESI-MS m / z:461.16[M+H] + .

[0186] 2-(2-(5-(4-acetylphenoxy)pentanoylamino)benzamido)benzoic acid (Z35)

[0187]

[0188] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-acetylphenoxy)pentanoic acid (6d), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z35: white solid, yield 52%, mp 103.2-106.4℃; 1 H NMR (400MHz, DMSO-d6) δ13.03(s,1H),10.82(s,1H),8.61(d,J=8.0Hz,1H),8.19(d ,J=8.2Hz,1H),8.06(dd,J=7.8,1.3Hz,1H),7.89(d,J=8.8Hz,2H),7.84(d,J=6.9Hz ,1H),7.60-7.50(m,2H),7.24(t,J=7.2Hz,1H),7.20–7.12(m,1H),7.00(d,J=8.9Hz ,2H),4.05(t,J=5.7Hz,2H),2.50(s,3H),2.43(t,J=6.7Hz,2H),1.82-1.68(m,4H); 13 C NMR(101MHz,DMSO-d6)δ196.71,171.45,170.31,166.96,162.96,141.07,138.65,133.35,132.51,131.70,130.90,1 30.20,128.41,124.55,123.98,123.36,122.46,120.38,116.05,114.69,67.99,36.91,28.37,26.85,21.98; ESI-MS m / z:497.1658[M+Na] + .

[0189] 2-(2-(4-(4-(tert-butyl)phenoxy)butyrylamino)benzamido)benzoic acid (Z36)

[0190]

[0191] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-(tert-butyl)phenoxy)butyric acid (6e), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z36: white solid, yield 76%, mp 87.3-89.5℃; 1H NMR(600MHz,DMSO-d6)δ13.79(s,1H),12.02(s,1H),10.59(s,1H),8.61(d,J=8.3Hz,1H),8 .09(d,J=8.2Hz,1H),8.04(dd,J=7.9,1.4Hz,1H),7.79(d,J=6.9Hz,1H),7.65–7.59(m,1H) ,7.60–7.51(m,1H),7.27(t,J=7.4Hz,1H),7.23-7.20(m,3H),6.80(d,J=8.8Hz,1H),6.80( d,J=8.8Hz,1H),3.94(t,J=6.4Hz,2H),2.60–2.43(m,2H),2.12–1.88(m,2H),1.22(s,9H); 13 C NMR (151MHz, DMSO-d6) δ171.20,170.18,166.90,156.71,143.05,141.16,138.15,134.43,132.52,131.65,129. 13,128.38,126.44,124.24,123.63,122.96,120.84,117.91,114.37,67.06,34.17,33.71,31.80,25.06; ESI-MS m / z:473.22[MH] - .

[0192] 2-(2-(5-(4-(tert-butyl)phenoxy)pentanoylamino)benzamido)benzoic acid (Z37)

[0193]

[0194] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-(tert-butyl)phenoxy)pentanoic acid (6f), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z37: white solid, yield 59%, mp 176.9-180.5℃; 1H NMR (600MHz, DMSO-d6) δ14.00(s,1H),11.05(s,1H),8.62(d,J=8.2Hz,1H),8.29(d,J=8.2H z,1H),8.06(d,J=7.4Hz,1H),7.89(d,J=7.6Hz,1H),7.53(t,J=7.8Hz,1H),7.48(t,J=7.7H z,1H),7.23(dd,J=16.5,8.1Hz,1H),7.12(t,J=7.3Hz,3H),6.82(d,J=8.7Hz,1H),6.82(d, J=8.7Hz,1H),3.93(t,J=5.5Hz,2H),2.43(t,J=6.6Hz,2H),1.77-1.71(m,4H),1.23(s,9H); 13 C NMR(151MHz,DMSO-d6)δ171.46,170.32,167.03,156.83,142.97,141.07,139.19,132.48,132.37,131.68,128.42, 126.44,123.71,123.08,121.99,119.99,114.36,67.48,37.18,34.18,31.82,28.59,22.13; ESI-MSm / z:487.22[MH] - .

[0195] 2-(2-(4-(4-ethylphenoxy)butyrylamino)benzamido)benzoic acid (Z38)

[0196]

[0197] According to the preparation method of compound Z1, compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z38: white solid, yield 52%, mp 137.0-140.4℃; 1H NMR (600MHz, DMSO-d6) δ12.15(s,1H),10.63(s,1H),8.61(dd,J=8.3,0.7Hz,1H),8.10(d,J=8.2Hz,1H),8.04(d d,J=7.9,1.5Hz,1H),7.80(dd,J=7.8,1.3Hz,1H),7.64–7.59(m,1H),7.58–7.53(m,1H),7.26(m,J=7.7,1.0Hz,1 H),7.23–7.19(m,1H),7.05(d,J=8.6Hz,1H),7.05(d,J=8.6Hz,1H),6.79(d,J=8.6Hz,1H),6.79(d,J=8.6Hz,1H ),3.94(t,J=6.4Hz,2H),2.51(t,J=5.1Hz,2H),2.49(q,J=7.2Hz,2H),2.01-1.97(m,2H),1.12(t,J=7.6Hz,3H); 13 C NMR (151MHz, DMSO-d6) δ171.21,170.19,166.91,157.02,141.14,138.22,136.08,134.27,132.52,131.65,129. 01,128.39,125.27,124.20,123.60,122.88,120.78,118.24,114.75,67.10,33.74,27.74,25.07,16.35; ESI-MS m / z:445.34[MH] - .

[0198] 2-(2-(4-(4-propionylphenoxy)butyrylamino)benzamido)benzoic acid (Z39)

[0199]

[0200] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-propionylphenoxy)butyric acid (6g), compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z39: white solid, yield 42%, mp 146.0-148.9℃; 1H NMR (400MHz, DMSO-d6) δ12.35(s,1H),10.69(s,1H),8.59(d,J=8.3Hz,2H),8.11(d,J=8.2 Hz,1H),8.04(d,J=7.7Hz,1H),7.88(d,J=8.7Hz,2H),7.81(d,J=7.6Hz,1H),7.64-7.52(m ,2H),7.26(t,J=7.5Hz,1H),7.19(t,J=7.5Hz,1H),6.98(d,J=8.7Hz,1H),4.08(t,J=6.3H z,2H),2.94(q,J=7.2Hz,2H),2.55–2.51(m,2H),2.11–1.95(m,2H),1.06(t,J=7.2Hz,3H); 13 C NMR(101MHz,DMSO-d6)δ199.21,171.10,170.24,166.89,162.68,141.12,138.22,134.03,132.50,131.65,130.48, 129.92,128.40,125.24,124.21,123.51,122.85,120.62,118.65,114.67,67.53,33.60,31.21,24.85,8.76; ESI-MS m / z:497.16[M+Na] + .

[0201] 2-(2-(5-(4-propionylphenoxy)pentanoylamino)benzamido)benzoic acid (Z40)

[0202]

[0203] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-propionylphenoxy)pentanoic acid (6h), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z40: white solid, yield 51%, mp 137.0-139.1℃; 1H NMR (400MHz, DMSO-d6) δ11.94(s,1H),10.56(s,1H),8.60(d,J=8.2Hz,1H),8.09(d,J=8.2Hz, 1H),8.03(dd,J=7.9,1.3Hz,1H),7.90(d,J=8.8Hz,2H),7.79(d,J=7.8Hz,1H),7.72–7.59(m, 1H),7.55(dd,J=11.4,4.2Hz,1H),7.18-7.30(m,2H),6.99(d,J=8.8Hz,2H),4.03(t,J=5.6Hz ,2H),2.96(q,J=7.2Hz,2H),2.41(t,J=6.7Hz,2H),1.82-1.65(m,4H),1.07(t,J=7.2Hz,3H); 13 C NMR(101MHz,DMSO-d6)δ199.24,171.49,170.15,166.92,162.80,141.10,138.13,134.47,132.53,131.63,130.50,129 .87,128.42,125.37,124.22,123.69,122.92,120.92,117.85,114.68,67.94,36.72,31.23,28.37,21.97,8.78; ESI-MS m / z:511.18[M+Na] + .

[0204] 2-(2-(4-(4-(Benzyloxy)phenoxy)butyrylamino)benzylamino)benzoic acid (Z41)

[0205]

[0206] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-(benzyloxy)phenoxy)butyric acid (6i), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z41: white solid, yield 47%, mp 158.1-160.6℃; 1H NMR (400MHz, DMSO-d6) δ12.74(s,1H),10.76(s,1H),8.61(d,J=8.3Hz,1H),8.15(d,J=8.2Hz,1H),8.06(d,J=7.7Hz,1H),7.83(d,J=7.6Hz,1H),7.62 -7.50(m,2H),7.45-7.28(m,5H),7.28-7.15(m,2H),6.89-6.81(m,4H),5. 00(s,2H),3.91(t,J=6.3Hz,2H),2.49(t,J=7.5Hz,2H),2.05–1.89(m,2H); 13 C NMR(101MHz,DMSO-d6)δ196.71,171.45,170.31,166.96,162.96,141.07,138.65,134.59,133.34,132.51,131.70,130.90,1 30.21,128.41,124.55,123.97,123.63,123.36,123.18,122.45,120.37,116.05,114.69,67.99,28.37,26.85,21.98; ESI-MS m / z:547.18[M+Na] + .

[0207] 2-(2-(5-(4-(Benzyloxy)phenoxy)pentanoylamino)benzamido)benzoic acid (Z42)

[0208]

[0209] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-(benzyloxy)phenoxy)pentanoic acid (6j), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z42: white solid, yield 58%, mp 128.6-131.4℃; 1H NMR(600MHz,DMSO-d6)δ12.27(s,1H),10.63(s,1H),8.61(d,J=8.1Hz,1H),8 .13(d,J=7.7Hz,1H),8.04(d,J=7.4Hz,1H),7.81(d,J=7.4Hz,1H),7.65-7.50 (m,2H),7.41(dd,J=23.8,6.8Hz,4H),7.35-7.15(m,3H),6.90(d,J=8.3Hz,2H ),6.82(d,J=8.3Hz,2H),5.02(s,2H),3.86(s,2H),2.40(s,2H),1.70(s,4H); 13 C NMR(151MHz,DMSO-d6)δ171.53,170.18,166.95,153.28,152.75,141.09,138.35,137.87,134.12,132.53,131.65,128.84,1 28.41,128.17,128.05,125.04,124.11,123.59,122.75,120.79,116.14,115.74,70.13,67.96,36.88,28.64,22.10; ESI-MS m / z:539.40[M+H] + .

[0210] 2-(2-(4-([1,1'-biphenyl]-2-yloxy)butyrylamino)benzamido)benzoic acid (Z43)

[0211]

[0212] According to the preparation method of compound Z1, compound 6a was replaced by 4-([1,1'-biphenyl]-2-oxy)butyric acid (6k), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z43: white solid, yield 58%, mp 167.8-169.5℃; 1H NMR (600MHz, DMSO-d6) δ14.18(s,1H),11.13(s,1H),8.63(d,J=8.2Hz,1H),8.31(d,J=8 .2Hz,1H),8.07(s,1H),7.91(d,J=5.6Hz,1H),7.57-7.45(m,4H),7.42(t,J=7.6Hz,2H) ,7.33–7.25(m,3H),7.22(t,J=7.0Hz,1H),7.13(t,J=6.5Hz,1H),7.08(d,J=8.3Hz,1H) ,7.01(t,J=7.4Hz,1H),4.03(t,J=6.2Hz,2H),2.50(t,J=1.6Hz,2H),2.01–1.94(m,2H); 13 C NMR(151MHz,DMSO-d6)δ171.03,167.06,155.79,141.04,139.26,138.63,132.53,132.22,130.88,130.43,129.7 2,129.27,128.39,127.23,123.72,123.49,123.10,121.88,121.31,119.89,113.21,67.47,34.00,25.04; ESI-MS m / z:493.31[MH] - .

[0213] 2-(2-(5-([1,1'-biphenyl]-2-oxy)pentanoylamino)benzamido)benzoic acid (Z44)

[0214]

[0215] According to the preparation method of compound Z1, compound 6a was replaced by 5-([1,1'-biphenyl]-2-oxy)pentanoic acid (6l), compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z44: white solid, yield 41%, mp 147.7-150.3℃; 1H NMR (600MHz, DMSO-d6) δ13.74(s,1H),11.99(s,1H),10.54(s,1H),8.59(d,J=8.3Hz,1H),8.11(d,J=8.2Hz,1H), 8.03(dd,J=7.9,1.4Hz,1H),7.85–7.76(m,1H),7.65–7.58(m,1H),7.55(dd,J=11.4,4.2Hz,1H),7.48(d,J=7.3Hz ,1H),7.48(d,J=7.3Hz,1H),7.35(t,J=7.7Hz,1H),7.35(t,J=7.7Hz,1H),7.33–7.23(m,4H),7.21(t,J=7.5Hz,1 H),7.06(d,J=8.2Hz,1H),7.01(t,J=7.4Hz,1H),3.95(d,J=5.5Hz,2H),2.34(t,J=6.7Hz,2H),1.70-1.64(m,4H); 13 C NMR(151MHz,DMSO-d6)δ171.49,170.14,166.95,155.91,141.09,138.62,138.30,134.39,132.56,131.62,130.86,130.41,129.69,1 29.24,128.36,128.31,127.17,125.10,124.14,123.69,122.82,121.24,120.93,118.02,113.23,67.84,36.67,28.50,21.98; ESI-MS m / z:507.18[MH] - .

[0216] 2-(2-(4-(2,4-di-tert-butylphenoxy)butyrylamino)benzamido)benzoic acid (Z45)

[0217]

[0218] According to the preparation method of compound Z1, compound 6a was replaced by 4-(2,4-di-tert-butylphenoxy)butyric acid (6m), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z45: white solid, yield 68%, mp 160.9-162.9℃; 1H NMR (600MHz, DMSO-d6) δ14.14(s,1H),11.16(s,1H),8.62(d,J=8.2Hz,1H),8.30(d,J=8 .2Hz,1H),8.06(d,J=6.0Hz,1H),7.90(d,J=6.8Hz,1H),7.54(t,J=7.7Hz,1H),7.46(t,J =7.7Hz,1H),7.22(dd,J=12.2,4.8Hz,2H),7.15–7.06(m,2H),6.82(d,J=8.5Hz,1H),4. 00(t,J=6.4Hz,2H),2.59(t,J=7.4Hz,2H),2.17–2.03(m,2H),1.35(s,9H),1.24(s,9H); 13 C NMR (151MHz, DMSO-d6) δ171.03,167.01,155.43,142.19,141.08,139.15,136.60,132.49,132.23,131.72,128.39,123. 88,123.79,123.34,123.04,122.03,119.87,112.28,67.19,35.02,34.33,31.90,30.28,25.45; ESI-MSm / z:529.40[MH] - .

[0219] 2-(2-(5-(2,4-di-tert-butylphenoxy)pentanoylamino)benzamido)benzoic acid (Z46)

[0220]

[0221] According to the preparation method of compound Z1, compound 6a was replaced by 5-(2,4-di-tert-butylphenoxy)pentanoic acid (6n), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z46: white solid, yield 60%, mp 167.6-168.7℃; 1H NMR (600MHz, DMSO-d6) δ13.77(s,1H),11.01(s,1H),8.62(d,J=8.2Hz,1H),8.2 9(d,J=8.2Hz,1H),8.06(d,J=6.3Hz,1H),7.89(d,J=6.7Hz,1H),7.53(t,J=7.7 Hz,1H),7.48(t,J=7.7Hz,1H),7.27–7.17(m,2H),7.17–7.05(m,2H),6.83(d,J =8.6Hz,1H),3.95(s,2H),2.45(s,2H),1.81(s,4H),1.32(s,9H),1.24(s,9H); 13 C NMR(151MHz,DMSO-d6)δ171.43,167.01,155.54,142.08,141.09,139.08,136.59,132.62,132.50,131.71,128.40,123 .84,123.79,123.31,123.15,122.12,120.09,112.18,67.43,37.11,35.01,34.33,31.91,30.21,28.91,22.40; ESI-MS m / z:543.21[MH] - .

[0222] 2-(2-(4-(Naphthalen-2-yloxy)butyrylamino)benzamido)benzoic acid (Z47)

[0223]

[0224] According to the preparation method of compound Z1, compound 6a was replaced by 4-(naphthalene-2-oxy)butyric acid (6o), compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z47: white solid, yield 56%, mp 162.1-163.9℃; 1H NMR(600MHz,DMSO-d6)δ13.74(s,1H),11.99(s,1H),10.62(s,1H),8.59(d,J=8.3Hz,1H), 8.11(d,J=8.2Hz,1H),8.02(dd,J=7.9,1.2Hz,1H),7.82-7.74(m,4H),7.63–7.52(m,2H), 7.43(t,J=7.4Hz,1H),7.33(t,J=7.3Hz,1H),7.29–7.24(m,2H),7.19(t,J=7.6Hz,1H),7. 13(dd,J=8.9,2.4Hz,1H),4.11(t,J=6.4Hz,2H),2.56(t,J=7.3Hz,2H),2.12-2.05(m,2H); 13 C NMR(151MHz,DMSO-d6)δ171.21,170.16,166.91,156.88,141.13,138.16,134.74,134.41,132.53,131.63,129.67,128.90,128.4 1,127.94,127.10,126.78,125.40,124.25,123.95,123.64,122.97,120.86,119.15,117.90,107.17,67.25,33.73,25.00; ESI-MS m / z:467.22[MH] - .

[0225] 2-(2-(5-(Naphthalen-2-yloxy)pentanoylamino)benzamido)benzoic acid (Z48)

[0226]

[0227] According to the preparation method of compound Z1, compound 6a was replaced by 4-(naphthalene-2-oxy)pentanoic acid (6p), and compound 5a was replaced by ethyl 2-(2-aminobenzamide)benzoate (5c), and the other conditions were the same. Compound Z48: white solid, yield 62%, mp 162.3-164.1℃; 1H NMR (400MHz, DMSO-d6) δ12.00(s,1H),10.58(s,1H),8.61(d,J=8.3Hz,1H),8.10(d,J=8 .2Hz,1H),8.04(d,J=7.2Hz,1H),7.86–7.73(m,4H),7.50-7.68(m,2H),7.44(t,J=7.4H z,1H),7.33(t,J=7.4Hz,1H),7.27(dd,J=8.9,5.1Hz,2H),7.20(t,J=7.5Hz,1H),7.13( dd,J=8.9,2.2Hz,1H),4.06(t,J=5.5Hz,2H),2.44(t,J=6.6Hz,2H),1.70-1.90(m,4H); 13 C NMR(101MHz,DMSO-d6)δ171.54,170.16,166.93,156.99,141.11,138.16,134.77,134.41,132.52,131.63,129.67,128.87,128.44,1 27.95,127.10,126.79,125.37,124.21,123.92,123.66,122.92,120.91,119.22,117.99,107.06,67.62,36.80,28.52,22.12; ESI-MS m / z:505.17[M+Na] + .

[0228] 2-(2-(5-(4-acetylphenoxy)pentanoylamino)-5-methylbenzamido)benzoic acid (Z49)

[0229]

[0230] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-acetylphenoxy)pentanoic acid (6d), and compound 5a was replaced by ethyl 2-(2-amino-5-methylbenzamide)benzoate (5d), and the other conditions were the same. Compound Z49: white solid, yield 58%, mp 175.9-178.7℃; 1H NMR (400MHz, DMSO-d6) δ13.75 (s, 1H), 11.87 (s, 1H), 10.43 (s, 1H), 8.59 (d, J = 8.1Hz, 1H),8.02(dd,J=7.9,1.5Hz,1H),7.94(d,J=8.3Hz,1H),7.89(d,J=8.8Hz,2H),7.68–7 .57(m,2H),7.37(d,J=8.4Hz,1H),7.21(t,J=7.6Hz,1H),6.99(d,J=8.8Hz,2H),4.02 (t,J=5.8Hz,2H),2.50(s,3H),2.38(t,J=6.8Hz,2H),2.34(s,3H),1.80–1.65(m,4H); 13 C NMR (101MHz, DMSO-d6) δ196.71,171.34,170.22,166.93,162.96,141.13,135.66,134.52,133.40,132.99,131.61,130. 90,130.22,128.66,125.42,123.65,123.06,120.88,117.70,114.68,67.97,36.66,28.37,26.85,21.99,20.86; ESI-MS m / z:511.18[M+Na] + .

[0231] 2-(5-methyl-2-(4-(4-propionylphenoxy)butyrylamino)benzamido)benzoic acid (Z50)

[0232]

[0233] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-propionylphenoxy)butyric acid (6g), compound 5a was replaced by ethyl 2-(2-amino-5-methylbenzamide)benzoate (5d), and the other conditions were the same. Compound Z50: white solid, yield 59%, mp 189.2-191.1℃; 1H NMR (400MHz, DMSO-d6) δ13.75(s,1H),11.93(s,1H),10.48(s,1H),8.58(d,J=8.3Hz,1H),8. 02(d,J=7.4Hz,1H),7.94(d,J=8.3Hz,1H),7.87(d,J=8.7Hz,2H),7.66–7.57(m,2H),7.37(d ,J=8.0Hz,1H),7.20(t,J=7.6Hz,1H),6.97(d,J=8.7Hz,2H),4.06(t,J=6.3Hz,2H),2.94(q, J=7.2Hz,2H),2.47(t,J=6.3Hz,2H),2.34(s,3H),2.06-1.98(m,2H),1.06(t,J=7.2Hz,3H); 13 C NMR(101MHz,DMSO-d6)δ199.22,170.96,170.23,166.89,162.69,141.17,135.60,134.44,133.47,132.97,131.62,130 .49,129.92,128.65,125.54,123.58,123.13,120.75,117.79,114.68,67.55,33.47,31.22,24.88,20.87,8.77; ESI-MS m / z:511.18[M+Na] + .

[0234] 5-Methyl-2-(4-methyl-2-(5-(4-propionylphenoxy)pentanoylamino)benzamido)benzoic acid (Z51)

[0235]

[0236] According to the preparation method of compound Z1, compound 6a was replaced by 4-(4-propionylphenoxy)butyric acid (6g), compound 5a was replaced by ethyl 2-(2-amino-4-methylbenzamido)benzoate (5e), ​​and the other conditions were the same. Compound Z51: white solid, yield 48%, mp 161.9-163.7℃; 1H NMR (400MHz, DMSO-d6) δ13.69(s,1H),11.89(s,1H),10.74(s,1H),8.46(d,J=8.7Hz,1H ),8.04(s,1H),7.90(d,J=8.7Hz,2H),7.83(s,1H),7.69(d,J=8.2Hz,1H),7.43(d,J=8.1 Hz,1H),7.07(d,J=7.7Hz,1H),6.99(d,J=8.5Hz,2H),4.04(t,2H),2.96(q,J=6.9Hz,2H) ,2.44-2.37(m,2H),2.35(s,3H),2.31(s,3H),1.82-1.65(m,4H),1.06(t,J=7.1Hz,3H); 13 C NMR(101MHz,DMSO-d6)δ199.24,171.42,170.19,166.88,162.81,142.86,138.67,134.86,132.78,131.70,130.50,12 9.87,128.25,124.67,122.74,121.41,120.95,114.70,67.96,36.96,31.23,28.36,22.00,21.75,20.73,8.79; ESI-MS m / z:539.21[M+Na] + .

[0237] 3-(2-(5-(4-acetylphenoxy)pentanoylamino)benzamido)-4-ethylbenzoic acid (Z52)

[0238]

[0239] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-acetylphenoxy)pentanoic acid (6d), and compound 5a was replaced by 3-(2-aminobenzamido)-4-ethylbenzoic acid ethyl ester (5f), and the other conditions were the same. Compound Z52: white solid, yield 38%, mp 137.3-139.8℃; 1H NMR (400MHz, DMSO-d6) δ13.10(s,1H),8.13(d,J=7.8Hz,1H),8.04(d,J=8.1Hz,1H),7.96( s,1H),7.88(dd,J=15.4,8.5Hz,3H),7.71(d,J=8.1Hz,1H),7.62(d,J=8.1Hz,1H),7.54(t ,J=7.5Hz,1H),6.93(d,J=8.7Hz,2H),4.00–3.98(m,2H),2.51(s,3H),2.46-2.35(m,2H), 2.35–2.19(m,2H),1.83(dd,J=14.7,7.4Hz,2H),1.77–1.62(m,2H),1.08(t,J=7.5Hz,3H); 13 C NMR(101MHz,DMSO-d6)δ196.71,166.99,162.82,161.72,156.65,147.68,146.26,136.34,135.23,130.90,130.75,130.60, 130.38,130.22,130.05,127.53,127.16,126.85,120.78,114.61,68.15,34.95,28.16,26.85,23.81,22.95,13.76; ESI-MS m / z:503.14[M+H] + .

[0240] 4-Ethyl-3-(2-(5-(4-propionylphenoxy)pentanoylamino)benzamido)benzoic acid (Z53)

[0241]

[0242] According to the preparation method of compound Z1, compound 6a was replaced by 5-(4-propionylphenoxy)pentanoic acid (6h), and compound 5a was replaced by 3-(2-aminobenzamido)-4-ethylbenzoic acid ethyl ester (5f), and the other conditions were the same. Compound Z53: white solid, yield 40%, mp 91.5-93.8℃; 1H NMR (400MHz, DMSO-d6) δ13.14(s,1H),8.13(dd,J=7.9,1.2Hz,1H),8.04(dd,J=8.0,1.7Hz,1 H),7.96(d,J=1.6Hz,1H),7.94–7.83(m,3H),7.71(d,J=8.0Hz,1H),7.62(d,J=8.1Hz,1H),7 .58–7.50(m,1H),6.93(d,J=8.9Hz,2H),4.05–3.91(m,2H),2.97(q,J=7.2Hz,2H),2.46-2.3 6(m,5.2Hz,2H),2.35–2.20(m,2H),1.90–1.77(m,2H),1.76–1.61(m,2H),1.10-1.06(m,6H); 13 C NMR(101MHz,DMSO-d6)δ199.24,166.99,162.67,161.72,156.65,147.68,146.26,136.34,135.23,130.75,130.60,130.51,13 0.38,130.05,129.88,127.53,127.15,126.85,120.78,114.62,68.12,34.95,31.23,28.17,23.81,22.96,13.76,8.78; ESI-MS m / z:517.13[M+H] + .

[0243] Example 3: Dual luciferase reporter gene assay analysis and cytotoxicity test

[0244] 1. Dual luciferase reporter gene assay analysis

[0245] HEK293T cells were routinely cultured at 37°C and 5% CO2 saturated humidity. One day before transfection, cells were seeded on a 96-well plate at 5,000 cells / well in DMEM medium containing 10% FBS and 1% penicillin-streptomycin; on the day of transfection, when the cell confluence reached 50%-60%, the old medium was aspirated, and then 100 μL of DMEM medium was added to each well, and the plate was placed in a 5% CO2, 37°C incubator; 0.15 μL of Polyethylenimine Linear was diluted with DMEM medium, and 50 ng of pRL-TK, pGL3-BASIC-FXR, and Pcdna3.1-NR1H4 were added respectively, with a final volume of 30 μL to obtain a transfection complex, which was allowed to stand at room temperature for 15 minutes, 30 μL of transfection complex solution was added to each well, and the 96-well plate was shaken to mix slightly; at 5% Incubate in a CO2, 37°C incubator for 6 hours, replace the medium containing the transfection complex with a fresh complete medium (containing FBS); after 24 hours of culture, remove the old medium, add the complete medium containing the drug and continue to culture in a 5% CO2, 37°C incubator for 24 hours. After the end, discard the original medium, add 30 μL of lysis buffer (Passive lysis Buffer) to each well, shake gently at room temperature for 15 minutes, and collect the cell lysate. Use UElandy's Dual luciferase reporter assay system (F6075L) to detect the firefly luciferase (Fluc) and Renilla luciferase (Rluc) signals of the sample. Read the results with the Envision microplate reader. Data were curve fitted using Prism 8.0 to calculate the EC 50 The results are shown in Table 1.

[0246] 2. Cell proliferation inhibition activity test

[0247] The cell proliferation inhibitory activity of the synthesized compounds was detected by MTT method. HepG-2 and L0-2 cells in the logarithmic growth phase were uniformly inoculated in 96-well plates. After the cells adhered to the wall, the drugs were given at concentrations of 400μM, 300μM, 250μM, 150μM, 100μM, 50μM, 25μM, and 12.5μM, respectively, and cultured in a cell culture incubator for 24h. The control group was added with an equal amount of DMSO. Then 20μL MTT solution was added to each well for reaction and incubated in the dark for 4h. After the incubation, the culture medium in the well was aspirated, 150μL DMSO was added to each well, and the crystals were fully dissolved by low-speed shaking. The blank group was the well without cells. The absorbance value of each well was measured at 490nm. The calculation formula of cell viability is: cell viability (%) = (OD to be tested-OD blank) / (OD control-OD blank) × 100.

[0248] FXR agonist activity test results and discussion: A: 0.1 μM <EC 50 <10μM; B:10μM <EC 50 <30 μM;

[0249] C:EC 50 >30μM.

[0250] Table 1 Compounds Z1-Z53 have FXR agonist activity and toxicity to HepG-2 and L0-2 cells

[0251]

[0252]

[0253]

[0254] The data are expressed as the geometric mean of at least three independent runs; OCA (obeticholic acid) and CDCA are positive control compounds, wherein OCA is a marketed FXR agonist, and CDCA is also a FXR agonist.

[0255] The results in Table 1 show that some compounds exhibit strong FXR agonist activity, which is comparable to that of the positive control compound OCA and superior to that of the positive control compound CDCA. They are basically non-toxic to HepG-2 and L0-2 cells, while the positive control drug OCA has a cytotoxicity IC of 2.34% to HepG-2 and L0-2 cells. 50 The cytotoxicity of compounds Z4 and Z5 was 257.50±1.63μM and 168.10±1.25μM, respectively, which was greater than that of most compounds synthesized by the present invention. 50 The values ​​were 1.35 μM and 2.23 μM, respectively, and they had no cytotoxicity to HepG-2 and L0-2 cells (IC 50 >100 μM).

[0256] Example 4: Oil Red O staining analysis, TG content determination, refer to Figure 1 .

[0257] The present invention uses 0.5 mM FFA to directly stimulate HepG-2 cells for 24 hours to construct a cell fatty degeneration model, and determines the effects of compounds Z4 and Z5 on lipid accumulation by Oil Red O staining and cell TG content.

[0258] from Figure 1A It can be seen that when the concentration is ≤300 μM, the survival rate of HepG-2 treated with Z4 and Z5 reaches more than 70.0%, and the cytotoxicity of compounds Z4 and Z5 is relatively small. However, at 300 μM, the survival rate of HepG-2 treated with OCA is only 1.2%, indicating that at a certain concentration, the cytotoxicity of compounds Z4 and Z5 is lower than that of OCA.

[0259] The present invention investigates the protective effect of Z4 and Z5 on lipid toxicity of high-fat HepG-2 cells. When the compound concentration is between 0 and 100 μg / mL, the survival rate of fatty HepG-2 cells treated with Z4, Z5 and OCA is basically above 100%, the fluctuation range of the survival rate is small, and there is no lipid toxicity ( Figure 1 B). In order to avoid interference from other factors and comprehensively consider cell survival rate and stability, this study selected 25, 50, and 75 μM as the low, medium, and high dose groups of Z4 and Z5 for subsequent research. The fat-soluble dye Oil Red O is more soluble in fat and can specifically stain with neutral fats such as TG in tissues and cells. After 24 hours of intervention with Z4 and Z5, the intracellular TG content was significantly reduced, the red lipid droplets were improved, and the absorbance value at 490 nm was also significantly reduced ( Figure 1 C, D, E).

[0260] Example 5: Evaluation of the in vivo anti-nonalcoholic steatohepatitis activity of compound Z4

[0261] 1. Compound Z4 improves obesity in NASH mice, see Figure 2 .

[0262] The present invention selects a high-fat diet and chemical induction. A high-fat diet accompanied by weekly intraperitoneal injection of CCl4 will aggravate liver damage in mice, thereby accelerating the development of fibrosis, effectively shortening the model cycle, and establishing a NASH mouse model accompanied by mild to moderate fibrosis. Obeticholic acid (OCA) was selected as a positive control drug. C57BL / 6J male mice aged 5 to 6 weeks were divided into a normal group (NCD group), a model group (NASH group), a low-dose group of Z4 (10 mg / kg), a medium-dose group of Z4 (20 mg / kg), a high-dose group of Z4 (40 mg / kg), and an OCA group (10 mg / kg), with 8 mice in each group. The experimental mice were fed a high-fat diet for 18 weeks. In the last five weeks, the model group and the drug group were intraperitoneally injected with 50 μL of 3.5% CCl4 solution twice a week, and the NCD group was intraperitoneally injected with 50 μL of olive oil twice a week and continuously gavaged with OCA (10 mg / kg) and Z4 (40 mg / kg, 20 mg / kg, 10 mg / kg) every day. The body weight and mental state of each mouse were recorded every week.

[0263] In order to determine the therapeutic effect of compound Z4 on NASH, the present invention first studied whether the administration of compound Z4 affects the systemic metabolic state. The NASH model was established by inducing a high-fat diet and CCl4 for 18 weeks and continued to be administered daily by gavage for the last 5 weeks ( Figure 2 A). During the drug intervention period, the mice in the NCD group were in good mental state and highly alert, while the mice in the NASH group were relatively confused, less alert, and less responsive. After treatment with Z4 and OCA, the mice's mental state improved, their activities increased significantly, and their body weight became significantly smaller. Compared with the NCD group, the mice in the NASH group gained weight significantly; after intraperitoneal injection of CCl4, the mice began to lose weight, while the mice treated with Z4 and OCA lost more weight than the NASH mice. Treatment with compounds Z4 and OCA limited the weight gain of NASH mice ( Figure 2 B). Morphological analysis of fat showed that the fat tissue of mice in the NASH group was hypertrophic compared with the fat cells of mice in the NCD group. H&E staining showed that in the same field of view, the number of fat cells was small, the shape was extremely irregular, the arrangement was disordered, the cell cross-sectional area was significantly increased, and the fat / body weight ratio was also significantly increased. After Z4 and OCA treatment, the fat tissue was significantly smaller, the number of fat cells was increased compared with the NASH group, the shape gradually became regular, the arrangement became neater, the cell cross-sectional area was significantly reduced, and the fat / body weight ratio was also significantly reduced ( Figure 2 C, D, E). Data are expressed as mean ± SEM. Compared with NASH, *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001. In conclusion, these data indicate that Z4 can effectively improve the obesity of NASH mice.

[0264] 2. Compound Z4 improves liver steatosis and liver damage in NASH mice, refer to Figure 3 .

[0265] The morphology and H&E staining of the liver showed that the liver of the mice in the NCD group was fresh, smooth and dense in color and texture, with complete hepatocyte morphology, equal size, clear outline and neatly arranged nuclei. The liver of the mice in the NASH group had many tiny milky white particles on the surface and the liver was rough and gray in appearance. The hepatocytes were enlarged, with many fat vacuoles, a large number of accumulated lipid droplets and disordered nuclei. After Z4 and OCA intervention, the appearance of the liver was significantly improved, the hepatocyte vacuoles were reduced, the outline became clear, and the nuclei gradually arranged neatly ( Figure 3 A). Organ index can indirectly reflect the degree of organ damage. Compared with the NCD group, the liver weight and liver index of mice in the NASH group increased significantly. After five weeks of Z4 and OCA intervention, the liver weight and liver index of mice decreased significantly ( Figure 3 B, C), and also significantly inhibited the upregulation of liver TG ( Figure 3 D).

[0266] Clinically, serum ALT and AST levels are sensitive indicators for detecting liver damage. Compared with the NCD group, the AST and ALT levels in the serum of mice in the NASH group were significantly increased. Z4 intervention significantly reduced the serum AST and ALT of mice, while OCA intervention only significantly reduced the serum AST of mice, and the ALT level only showed a downward trend, with no significant difference ( Figure 3 E, F). Long-term high blood lipid levels will cause fat accumulation in the liver, which will cause long-term lipotoxicity, activate the body's oxidative pathways, produce a large number of reactive oxygen species (ROS), and cause oxidative stress in the body. Catalase (CAT) and glutathione peroxidase (GSH-Px) are important antioxidant enzymes in the body and play an important role in the body's redox balance. Malondialdehyde (MDA) is a product of lipid peroxidation in the body and is a sensitive indicator for detecting free radical damage in the body. Treatment with compound Z4 significantly increased liver CAT and serum GSH-Px, and significantly reduced liver MDA ( Figure 3 G, H, I, J). Data are expressed as mean ± SEM. Compared with the NASH group, *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001. In conclusion, these data indicate that Z4 treatment can reduce hepatic steatosis and liver damage in NASH mice, increase the activity of antioxidant enzymes in the body, and enhance the body's antioxidant capacity.

[0267] 3. The effect of compound Z4 on improving lipid metabolism disorder in NASH mice, refer to Figure 4 .

[0268] from Figure 4 It can be seen that compared with the NCD group, the TC, TG and LDL-C levels of the NASH group mice serum were significantly increased, and the HDL-C level showed a downward trend, but there was no significant difference. After Z4 intervention, the TC, TG, and LDL-C levels of the mice were improved to varying degrees, among which the TC and TG levels were significantly reduced, the LDL-C content was reduced, and the HDL-C content showed an upward trend, but there was no significant difference ( Figure 4 A, B, C, D), data are expressed as mean ± SEM, compared with NASH, *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001. These results show that Z4 can well regulate the blood lipid levels in NASH mice and improve dyslipidemia, and its effect on improving dyslipidemia is comparable to that of the positive control compound OCA.

[0269] 4. Example 6: Pharmacokinetic analysis of compound Z4, refer to Figure 5 .

[0270] Male C57BL / 6J mice were selected to conduct pharmacokinetic and tissue distribution studies on compound Z4. The results showed that the oral bioavailability of compound Z4 was 61.2%, T 1 / 22.5h, T max The AUC of oral and intravenous administration is 0.25h. 0-last They are 2593.4 and 848.0 ng / mL*h respectively ( Figure 5 A). 0.5 hours after oral administration, the concentration of compound Z4 in the liver was 6230 ng / mL, significantly higher than that in plasma and small intestine. 2 hours after administration, the drug concentration in the small intestine was slightly higher than that in the liver. 8 hours after administration, the drug concentration in the liver and small intestine was lower than 1500 ng / mL ( Figure 5 B), data are presented as mean ± SEM. In conclusion, compound Z4 exhibits good pharmacokinetic properties.

[0271] In conclusion, the preferred compound Z4 of the present invention has strong agonist activity on FXR, high in vivo biological activity, good pharmacokinetic properties, and good application prospects.

[0272] Example 7: Acute toxicity of compound Z4 in C57BL / 6J mice

[0273] During the experiment, it was found that after a single oral administration of compound Z4 (3000mg / kg, 1500mg / kg, 750mg / kg, 375mg / kg, 187.5mg / kg, 93.75mg / kg, 40mg / kg), the mice were fed conventionally and observed for 14 days without any toxic side effects. All mice survived, and the lethal range of compound Z4 could not be measured after oral administration. After 14 days of autopsy, there was no significant difference in the weight of mice in each dosage group. Compared with the 0mg / kg group, the liver index and heart index of mice in the 375mg / kg dosage group increased significantly, but there was no significant effect on other organ indexes, and the AST and ALT activities in serum increased significantly. The liver index of mice in the 187.5mg / kg dosage group increased significantly, and other dosage groups had no significant effect on organ indexes, AST and ALT activities in serum. After oral administration of 1500 mg / kg, 750 mg / kg, 70 mg / kg, 40 mg / kg, 20 mg / kg, and 0 mg / kg of the positive compound OCA, the mice in the oral administration of 1500 mg / kg and 750 mg / kg groups all died one day after oral administration, and the kidney index of the mice in the oral administration of 70 mg / kg and 40 mg / kg groups, and the activities of AST and ALT in the serum of the mice increased significantly, showing toxic reactions; the activities of AST and ALT in the serum of the mice in the oral administration of 20 mg / kg group increased significantly, and the results are shown in Table 2. In short, compared with the positive compound OCA, compound Z4 is less toxic and safer in vivo.

[0274] Table 2 Acute toxicity of compounds Z4 and OCA in C57BL / 6J mice

[0275]

[0276] Note: Data are expressed as mean ± SEM. Compared with the 0 mg / kg group, * is significantly different (P < 0.05); ** is extremely significantly different (P < 0.01); *** is extremely significantly different (P < 0.001).

Claims

1. An aryloxyaminobenzoic acid derivative, characterized in that: Its structural formula is shown in formula (I), in: Ar is a substituted phenyl or naphthyl group selected from 4-ethylphenyl, 4-tert-butylphenyl, 2,4-di-tert-butylphenyl, 4-acetylphenyl, 4-propionylphenyl, 4-benzyloxyphenyl, biphenyl, and naphthyl; n is independently selected from 2 and 3; R1 and R2 are each independently selected from hydrogen, methyl, and ethyl; R1, R2 and the -COOH group are located at any position of the benzene ring.

2. The aryloxyaminobenzoic acid derivative according to claim 1, characterized in that: The derivative represented by formula (I) is selected from the following compounds:

3. The method for preparing the aryloxyaminobenzoic acid derivatives according to claim 1, characterized in that: The preparation route of the derivative represented by formula (I) is as follows: Wherein, R1 and R2 in the compounds represented by formula (II), formula (III), formula (IV), compounds 5a-f and formula (VI) and the compound represented by formula (I) have the same definition; The compounds represented by compounds 6a-p, the compounds represented by formula (VI) and the compounds represented by formula (I) have the same definitions as Ar and n; The preparation method comprises the following steps: Step a. Nitrobenzoic acid and its different derivatives II and ethyl aminobenzoate and its different derivatives III are subjected to an amidation reaction in a suitable solvent and a condensation reagent to prepare an intermediate compound IV; Step b. The compound represented by formula IV is subjected to reduction reaction in an organic solvent and a reducing agent to obtain compounds 5a-f; Step c. Compounds 5a-f are subjected to amidation reaction with aromatic oxygen carboxylic acid derivatives 6a-p respectively to generate aromatic oxygen aminobenzoic acid ester derivatives shown in formula VI; Step d: The compound represented by formula VI is hydrolyzed in an organic solvent under alkaline conditions, and then subjected to an acidification reaction to prepare a derivative represented by formula (I).

4. The method for preparing the aryloxyaminobenzoic acid derivatives according to claim 3, characterized in that: The molar ratio of nitrobenzoic acid and its different derivatives II to ethyl aminobenzoate and its different derivatives III described in step a is 1:1.1, and the reaction temperature is 30-45°C; The appropriate solvent in step a is selected from N,N-dimethylformamide, tetrahydrofuran, dichloromethane, chloroform, and dimethyl sulfoxide; The condensation reagent described in step a is selected from one of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N-diisopropylethylamine, 1-ethyl-3-(3-dimethylpropylamine)carbodiimide hydrochloride, 1-hydroxybenzotriazole, and dicyclohexylcarbodiimide.

5. The method for preparing the aryloxyaminobenzoic acid derivatives according to claim 3, characterized in that: The molar ratio of the compound represented by formula (IV) described in step b to the reducing agent is 1:5, and the reaction temperature is room temperature to 45°C; The organic solvent in step b is selected from ethanol, methanol, tetrahydrofuran and dichloromethane; The reducing agent described in step b is selected from zinc powder or iron powder; The method of step b, or reducing the compound represented by formula IV in ethanol with zinc powder to obtain derivatives 5a-f; Alternatively, the compound represented by formula IV is subjected to hydrogenation reduction in a hydrogen atmosphere with 5% Pd / C as a catalyst to prepare compounds 5a-f.

6. The method for preparing the aryloxyaminobenzoic acid derivatives according to claim 3, characterized in that: The molar ratio of compound 5a-f to compound 6a-p in step c is 1:1.

3.

7. The method for preparing the aryloxyaminobenzoic acid derivatives as claimed in claim 3, characterized in that: The molar ratio of the compound represented by formula (VI) in step d to the base is 1:1.1; The organic solvent in step d is selected from ethanol, methanol and tetrahydrofuran; The base described in step d is selected from sodium hydroxide or potassium hydroxide; The acidification reaction conditions in step d are to adjust the pH to 1-2 with 1 mol / L hydrochloric acid solution and react at 0-5°C.

8. Use of the derivative represented by formula (I) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof as a farnesoid X receptor modulator in the preparation of a medicament for treating farnesoid X receptor-related diseases according to claim 1, characterized in that: The application is specifically in the preparation of a drug for treating at least one of cholestatic liver disease, non-alcoholic fatty liver disease, hyperlipidemia, diabetes, obesity and atherosclerosis.

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