A class of pentacyclic triterpenoid derivatives and their medical uses

Abstract

The present invention discloses a class of pentacyclic triterpenoid saponins and their derivatives for use in preventing and treating myocardial hypertrophy; belonging to the field of medicinal chemistry; the pentacyclic triterpenoid derivatives or pharmaceutically acceptable salts thereof are represented by general formula (I): the pentacyclic triterpenoid derivatives of the present invention have a good binding effect on GATA6, a potential intervention target for myocardial hypertrophy, can significantly prevent and reverse myocardial cell hypertrophy in vitro and myocardial hypertrophy in vivo, and have potential medicinal value.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and relates to a class of pentacyclic triterpene derivatives and uses thereof, and in particular to a class of pentacyclic triterpene derivatives and uses thereof in preparing drugs for preventing and treating hypertrophic cardiomyopathy. Background Art

[0002] Cardiac hypertrophy (CH) is a common clinical pathological phenomenon that often occurs in various heart diseases, including hypertension, valvular heart disease, and myocardial infarction. It serves as a compensatory mechanism for increased cardiac load and is an independent predictor of mortality in patients with cardiovascular disease. As the disease progresses, the hypertrophic myocardium gradually becomes unable to withstand the increased load due to factors such as increased oxygen consumption, decreased adrenergic receptor density, and decreased contractility. This leads to decreased cardiac output, decompensated cardiac function, and symptoms of congestive heart failure. This also increases the susceptibility to complications such as arrhythmias and thromboembolism. Therefore, CH and HF can be considered two distinct stages of the same disease process. The pathogenesis of CH involves complex signaling pathway activation and pathological changes, which are currently incompletely understood, and clinical treatment responses are suboptimal. Therefore, clarifying the pathological mechanisms of CH and identifying new therapeutic targets and drugs for prevention and treatment are of great importance. Summary of the Invention

[0003] Purpose of the invention: In response to the above problems, the present invention discovered that GATA6 plays an important regulatory function in cardiomyocyte hypertrophy and can serve as a potential target for drug intervention, and provides a class of pentacyclic triterpene derivatives or pharmaceutically acceptable salts thereof, and their use in the preparation of drugs for the treatment of myocardial hypertrophy; another object of the present invention is to provide the use of the pentacyclic triterpene derivatives or pharmaceutically acceptable salts thereof in the preparation of drugs for the treatment of myocardial hypertrophy.

[0004] The technical solution of the present invention is: a class of pentacyclic triterpene derivatives or pharmaceutically acceptable salts thereof according to the present invention, wherein the chemical structural formula of the derivative is shown in formula (I):

[0005]

[0006] wherein R1 is H or OH; R2 is -CH2OH or -COOH; R3 is H or OH;

[0007] R4 is

[0008]

[0009] Furthermore, the pentacyclic triterpenoid saponin compound or a pharmaceutically acceptable salt thereof is selected from the following compounds:

[0010]

[0011]

[0012]

[0013]

[0014]

[0015]

[0016]

[0017] One object of the present invention is to provide a potential therapeutic target for myocardial hypertrophy. The present invention finds that GATA6 plays an important regulatory function in cardiomyocyte hypertrophy and can be used as a potential intervention target for the treatment of myocardial hypertrophy. Another object of the present invention is to provide the use of the pentacyclic triterpene derivative or pharmaceutically acceptable salt, and its pharmaceutical composition in the preparation of a drug for preventing or treating myocardial hypertrophy.

[0018] The beneficial effects of the present invention are: the pentacyclic triterpene derivatives in the present invention have a good binding effect on GATA6, a potential intervention target for myocardial hypertrophy, can significantly prevent and reverse myocardial cell hypertrophy in vitro and myocardial hypertrophy in vivo, and have potential medicinal value. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a schematic diagram of improving myocardial hypertrophy in mice by knocking down GATA6 in heart tissue in an embodiment of the present invention. DETAILED DESCRIPTION

[0020] Example 1, Synthesis of Compound 1-a:

[0021]

[0022] Compound 1 was purchased from a pharmaceutical technology company. 1 (100 mg, 0.15 mmol) was dissolved in methanol (2 mL), followed by the addition of NaOH solution (0.12 mL, 5 mol / L). The temperature was raised to 80°C and the reaction was allowed to react for 4 hours. After completion of the reaction, as monitored by LC-MS, 2 mL of water was added to the reaction system, and the pH of the mixture was adjusted to 3 with HCl solution (2 mol / L). A large amount of light yellow solid precipitated. The mixture was filtered and dried to obtain light yellow solid 1a in a 90% yield. NMR (500MHz, DMSO-d6) 1Hδ (ppm): 11.83 (s, 1H), 5.17 (t, J = 3.7Hz, 1H), 4.40 (t, J = 5.2Hz, 1H), 4.15 (d, J = 4.9Hz, 1 H),3.76(s,1H),3.45(dd,J=10.6,5.4Hz,1H),3.08(dd,J=10.5,5.0Hz,1H),2.37(s,1H),1.96–1.80(m,3H),1.69 (td,1H),1.64–1.57(m,3H),1.55–1.46(m,6H),1.40(dt,J=15.2,5.3Hz,3H),1.30(s,3H),1.28–1.24(m,2H),1.2 0–1.15m,(2H),1.15–1.11(m,2H),1.08(s,3H),0.88(s,3H),0.85(d,J=6.6Hz,3H),0.71(s,3H),0.55(s,3H).13C NMR(126MHz,DMSO-d6)δ(ppm):178.93,138.60,126.86,71.61(2C),70.4 1,64.51,53.18,46.88,46.71,46.45,41.83,41.39,41.13,39.28,38.00, 37.26,36.25,32.27,28.10,26.56,26.40,25.92,25.19,24.02,23.14,17 .59,16.62,16.27,15.47,12.61.ESI-MS(m / z)488.2[MH]-,490.7[M+H]+.

[0023] Example 2, Synthesis of Compound 2:

[0024]

[0025] Compound 1a (200 mg, 0.41 mmol) was dissolved in DMSO (2 mL), followed by the addition of HATU (172 mg, 0.45 mmol) and triethylamine (0.08 mL, 0.62 mmol). The mixture was stirred at room temperature for 0.5 h. Piperazine (105 mg, 1.23 mmol) was then added, and stirring continued at room temperature for 2 h. After completion of the reaction, as monitored by LC-MS, 5 mL of water was added to the reaction system, resulting in the precipitation of a large amount of solid. The residue was filtered, and the filter cake was washed with water (5 mL x 2). The crude product was then dried and purified by column chromatography (CH2Cl2:MeOH = 40:1) to afford pure white solid 2 in an 80% yield. 1 H NMR(400MHz,C5D5N)δ(ppm):5.58(s,1H),5.18(s,1H),4.40–4.15(m,2H),3.86–3.63(m,5H),3.41–3.20(m,2H ),3.12–2.86(m,4H),1.69(s,3H),1.44(s,3H),1.14(d,J=6.8Hz,3H),1.12(s,3H),1.08(s,3H),1.05(s,3H); 13 C NMR (126MHz, C5D5N) δ (ppm): 176.21, 140.65, 128.46, 74.08, 73.63, 68.46, 57.19, 50.06, 49.24, 48.39, 47.75, 47.26, 43.43, 42.82, 42.67, 40 .76,39.42,37.78,35.90,34.02,29.69,28.22,28.13,27.34,25.09,24.60,19.34,17.73,17.30,16.65,13.67; ESI-MS(m / z)602.1[M+HCOOH] - ,558.1[M+H] + .

[0026] Example 3, synthesis of compound 3:

[0027]

[0028] Compound 1a (200 mg, 0.41 mmol) was dissolved in DMSO (2 mL), followed by the addition of HATU (172 mg, 0.45 mmol) and triethylamine (0.08 mL, 0.62 mmol). The mixture was stirred at room temperature for 0.5 h. 1,4-Diazacyclohexane (123 mg, 1.23 mmol) was then added, and stirring continued at room temperature for 2 h. After completion of the reaction, as monitored by LC-MS, 5 mL of water was added to the reaction system, resulting in the precipitation of a large amount of solid. The residue was filtered, and the filter cake was washed with water (5 mL x 2). The crude product was then dried and purified by column chromatography (CH2Cl2:MeOH = 40:1) to afford pure white solid 3 in a 70% yield. 1 H NMR(500MHz,DMSO-d6)δ(ppm):5.10(s,1H),4.39(t,J=5.1Hz,1H),4.14(d,J=4.9Hz,1H),3.79(s,1H),3.56–3.37(m,5H),3.34(s,1H),3 .07(dd,J=10.5,1H),2.81(s,1H),2.63(s,1H),2.23(s,4H),2.14(s,3H),1.95–1.76(m,3H),1.65–1.55(m,3H),1.51(d,J=3.3Hz,1H),1. 49(s,2H),1.47-1.44(m,2H),1.39(d,J=12.7Hz,2H),1.28(s,3H),1.25(d,J=3.8Hz,1H),1.22(d,J=3.2Hz,1H),1.18–1.15(m,1H),1.15– 1.11(m,2H),1.09(s,3H),0.95–0.88(m,2H),0.87(s,3H),0.84(d,J=6.8Hz,3H),0.62(s,3H),0.54(s,3H).ESI-MS(m / z)616.2[M+HCOOH] - ,572.1[M+H] + .

[0029] Example 4: Synthesis of Compound 4:

[0030]

[0031] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with N-methylpiperazine (123 mg, 1.23 mmol) to obtain pure white solid powder 4 in a yield of 75%. 1H NMR(500MHz,DMSO-d6)δ(ppm):5.08(s,1H),4.41(s,1H),4.15(s,1H),3.77(s,1H),3.43(dd,J=11.6,5.0Hz,4H), 3.06(d,J=10.4Hz,1H),2.80–2.76(m,1H),2.66(s,2H),1.92(s,1H),1.88(t,J=5.3Hz,1H),1.86–1.77(m,2H),1.6 4–1.56(m,6H),1.52–1.44(m,6H),1.38(d,J=12.8Hz,3H),1.28(s,3H),1.26–1.19(m,3H),1.17–1.12(m,3H),1.1 0(s,1H),1.08(s,3H),0.86(s,3H),0.84(d,J=6.8Hz,5H),0.65(s,3H),0.53(s,3H).ESI-MS(m / z)616.2[M+HCOOH] - ,572.1[M+H] + .

[0032] Example 5, Synthesis of Compound 5:

[0033]

[0034] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with 4-amino-piperidine (123 mg, 1.23 mmol) to obtain pure white solid powder 5 with a yield of 80%. 1H NMR (500MHz, DMSO-d6) δ (ppm): 6.77 (d, J = 7.8Hz, 1H), 5.17 (d, J = 3.7Hz, 1H), 4.39 (s, 1H), 4.13 (d, J = 4.9Hz, 1H), 3.70 (s, 1H), 3.54–3.47 (m, 1H) ),3.47–3.39(m,2H),3.07(d,J=10.4Hz,1H),2.89(d,J=12.1Hz,2H),2 .44(s,2H),2.42–2.38(m,2H),1.94–1.84(m,2H),1.83-1.77(m,1H),1. 68–1.61(m,2H),1.61–1.54(m,4H),1.53(d,J=2.6Hz,1H),1.49(d,J=12.5Hz,5H),1.46(d,J=4.3Hz,1H),1.37(dd,J=12.7,5.1Hz,2H),1.28(s ,3H),1.27–1.23(m,3H),1.21-1.18(m,1H),1.13(d,J=13.1Hz,3H),1.0 8(s,3H),0.86(s,3H),0.83(d,J=6.7Hz,3H),0.65(s,3H),0.53(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):176.22,139.40,127.08,72.43(2C),70.90,6 5.00,53.08,47.22,47.02,46.95,46.86,45.66,42.30(2C),41.74,41.55(2C) ),38.51,38.41,36.71,33.26,32.95,28.35,27.06,26.98,26.58,24.85,24 .48,23.65,18.05,17.24,16.77,15.95,13.11.ESI-MS(m / z)616.1[M+HCOOH] - ,572.1[M+H] + .

[0035] Example 6, Synthesis of Compound 6:

[0036]

[0037] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with N-methylpiperidin-4-amine (140 mg, 1.23 mmol) to obtain pure white solid powder 6 in a yield of 70%. 1H NMR (500MHz, DMSO-d6) δ (ppm): 6.77 (d, J = 7.7Hz, 1H), 5.16 (t, J = 3.7Hz, 1H), 4.38 (t,J=5.2Hz,1H),4.13(d,J=5.0Hz,1H),3.70(s,1H),3.45-3.40(m,3H),3.07(dd ,J=10.5,5.0Hz,1H),2.69(t,J=4.1Hz,1H),2.67(s,1H),2.45(s,1H),2.11(s,3H ),1.92-1.87(m,1H),1.85(s,1H),1.82(d,J=3.1Hz,1H),1.82–1.79(m,1H),1.64( d,J=4.9Hz,1H),1.59(d,J=4.4Hz,1H),1.57(s,2H),1.55(s,1H),1.51(d,J=7.4H z,3H),1.48(d,J=3.9Hz,2H),1.47–1.41(m,4H),1.40(d,J=3.8Hz,1H),1.35(d,J =4.4Hz,1H),1.28(s,3H),1.23(s,2H),1.15(t,J=3.2Hz,1H),1.12(s,1H),1.08( s,3H),0.86(s,3H),0.83(d,J=6.7Hz,5H),0.80(s,1H),0.64(s,3H),0.53(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):175.94,138.87,126.59,71.91(2C),70.40,64.5 0,54.62,54.56,52.59,46.71,46.44,46.38,45.97,45.79,41.79,41.24,41.04 ,38.00,37.88,36.20,32.43,31.48,31.12,27.84,26.55,26.47,26.07,24.33, 23.98,23.14,17.54,16.68,16.25,15.44,12.59.ESI-MS(m / z)630.1[M+HCOOH] - ,586.1[M+H] + .

[0038] Example 7, Synthesis of Compound 7:

[0039]

[0040] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with piperidin-4-ylmethylamine (140 mg, 1.23 mmol) to obtain pure white solid powder 7 in 80% yield. 1 H NMR(500MHz,DMSO-d6)δ(ppm):7.13(t,J=5.7Hz,1H),5.18(t,J=3.7Hz,1H),4.41(s,1H),4.16 (s,1H),3.74(s,1H),3.43(dd,J=11.4,4.8Hz,2H),3.07(d,J=10.4Hz,1H),2.92–2.88(m,2H), 2.83–2.80(m,1H),2.77–2.74(m,1H),2.43(s,1H),2.39–2.33(m,2H),1.92–1.87(m,1H),1.82 –1.78(m,1H),1.65(d,J=4.8Hz,1H),1.63(d,J=4.1Hz,1H),1.60(d,J=4.6Hz,1H),1.57(d,J=3 .7Hz,1H),1.56–1.54(m,1H),1.53–1.50(m,3H),1.49(s,1H),1.49–1.47(m,2H),1.47–1.45(m ,2H),1.44(d,J=4.2Hz,1H),1.39(d,J=4.3Hz,1H),1.37(s,1H),1.28(s,3H),1.24(t,J=3.6Hz ,1H),1.20(d,J=3.2Hz,1H),1.12(t,J=6.9Hz,2H),1.09(s,1H),1.08(s,3H),0.93(s,1H),0.9 0(s,1H),0.86(s,3H),0.83(d,J=6.7Hz,J=6.7Hz,4H),0.78(s,1H),0.62(s,3H),0.53(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):178.22,140.47,128.13,73.40(2C),71.87,65. 98,54.23,48.18,48.09,47.93,47.12,46.58,43.28,42.71,42.49,39.58,39. 45,37.68(2C),37.17,33.85,32.26,29.32,28.01,27.93,27.56,26.10,25.51 ,24.60,19.03,18.11,17.73,16.88,14.07,1.54.ESI-MS(m / z)630.2[M+HCOOH]- ,586.1[M+H] + .

[0041] Example 8, Synthesis of Compound 8:

[0042]

[0043] Reaction and post-processing operations such as replacing 3,1,4-diazacyclohexane with N 1 ,N 1 -dimethylethane-1,2-diamine (108 mg, 1.23 mmol) to obtain pure white solid powder 8, yield: 80%. 1 H NMR(500MHz,DMSO-d6)δ(ppm):6.94(t,J=5.5Hz,1H),5.18(t,J=3.7Hz,1H),4.39(t,J=5.1Hz,1H),4.14(d,J=4.9Hz,1H),3.75(s,1H),3.45-3.41(m,1 H),3.10-3.05(m,2H),3.01–2.96(m,1H),2.54(dd,J=13.5,4.3Hz,1H),2.3 6(s,1H),2.23(t,J=6.9Hz,2H),2.12(s,5H),1.92–1.87(m,1H),1.86-1.82 (m,1H),1.65–1.61(m,1H),1.61–1.54(m,3H),1.53-1.51(m,2H),1.48(d,J =4.0Hz,1H),1.47–1.42(m,2H),1.40(s,2H),1.36(d,J=4.3Hz,1H),1.28(s ,3H),1.25–1.21(m,2H),1.14(s,1H),1.11(d,J=11.6Hz,3H),1.08(s,3H), 0.87(s,3H),0.83(d,J=6.7Hz,4H),0.80(s,1H),0.64(s,3H),0.53(s,3H). 13C NMR(126MHz,DMSO-d6)δ(ppm):176.54,138.60,126.68,71.73(2C),70.2 1,64.31,57.73(2C),52.78,46.53,46.38,46.25,45.01(4C),41.62,41.1 5,40.88,37.81,36.74,36.01,32.11,27.63,26.32,25.89,24.57,23.79 ,22.96,17.36,16.34,16.08,15.24,12.40.ESI-MS(m / z)604.1[M+HCOOH] - ,560.0[M+H] + .

[0044] Example 9, Synthesis of Compound 9:

[0045]

[0046] Reaction and post-processing operations such as replacing 3,1,4-diazacyclohexane with N 1 -methylethane-1,2-diamine (91 mg, 1.23 mmol) to obtain pure white solid powder 9, yield: 75%. 11H NMR (500 MHz, MeOD) δ (ppm): 5.36 (t, J = 3.7 Hz, 1H), 3.61 (dd, J = 11.5, 4.6 Hz, 1H), 3.53 (d, J = 10.9 Hz, 1H), 3.29 (s, 1H), 3.19–3.14 (m, 1H), 2.69 (d, J = 4.6 Hz, 1H), 2.64 (t, J = 6.5 Hz, 3H), 2.47 (s, 1H), 2.38 (s, 3H), 2.02–1.99 (m, 1H), 1.97 (t, J = 3.6 Hz, 1H), 1.74 (d, J = 3.2 Hz, 1H), 1.73–1.71 (m, 1H), 1.70 (d, J = 3.7 Hz, 1H), 1.67 (t, J = 3.2 Hz, 2H), 1.65 (s, 1H), 1.63–1.61 (m, 1H), 1.60 (d, J = 1.9 Hz, 1H), 1.55 (d, J = 4.4 Hz, 1H), 1.52 (d, J = 4.1 Hz, 1H), 1.50–1.48 (m, 1H), 1.44 (s, 1H), 1.41 (s, 1H), 1.38 (d, J = 3.2 Hz, 1H), 1.35 (s, 3H), 1.27 (d, J = 3.0 Hz, 1H), 1.23 (d, J = 3.0 Hz, 1H), 1.22 (s, 1H), 1.21 (s, 3H), 1.18 (d, J = 2.2 Hz, 1H), 1.16 (d, J = 2.3 Hz, 1H), 1.01 (d, J = 3.6 Hz, 1H), 0.99 (s, 1H), 0.97 (s, 3H), 0.96 (s, 1H), 0.93 (d, J = 6.6 Hz, 4H), 0.75 (s, 3H), 0.71 (s, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ (ppm): 176.87, 138.86, 126.78, 71.91 (2C), 70.40, 64.50, 52.84, 50.52, 46.71, 46.59, 46.44, 41.80, 41.30, 41.03, 38.49, 38.00, 36.20, 35.78 (2C), 32.30, 27.84, 26.54, 26.48, 26.07, 24.68, 24.01, 23.14, 17.54, 16.49, 16.26, 15.41, 12.59. ESI-MS (m / z) 590.0 [M+HCOOH] - , 546.1 [M+H] + .

[0047] Example 10, Synthesis of Compound 10:

[0048]

[0049] Reaction and post-processing operations such as replacing 3,1,4-diazacyclohexane with N 1 -ethylethane-1,2-diamine (109 mg, 1.23 mmol) was added to obtain pure white solid powder 10 with a yield of 75%. 1 H NMR(500MHz,DMSO-d6)δ(ppm):7.05(t,J=5.5Hz,1H),5.20(t,J=3.6Hz,1H),4.39( t,J=5.1Hz,1H),4.14(d,J=4.9Hz,1H),3.74(s,1H),3.45–3.41(m,2H),3.09–3.05( m,2H),3.00(dd,J=12.8,6.1Hz,1H),2.55–2.53(m,1H),2.52(s,1H),2.39(s,1H),1 .90(dd,J=6.6,3.5Hz,1H),1.84(dd,J=11.2,3.5Hz,1H),1.62–1.57(m,4H),1.55(s ,1H),1.53–1.51(m,2H),1.48(s,2H),1.46(t,J=5.1Hz,2H),1.42(s,1H),1.40(d,J =3.6Hz,1H),1.38(d,J=4.1Hz,1H),1.28(s,3H),1.27–1.25(m,1H),1.24(d,J=3.4H z,1H),1.21(d,J=3.3Hz,1H),1.14(s,1H),1.12(s,1H),1.10(s,1H),1.08(s,3H),0 .99(t,J=7.1Hz,4H),0.86(s,3H),0.83(d,J=6.8Hz,4H),0.63(s,3H),0.53(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):176.91,138.87,126.77,71.92(2C),70.38, 64.49,52.82,48.10,46.70,46.61,46.43,43.10,41.80,41.29,41.03,38.7 5,38.00,36.19(2C),32.27,27.83,26.53,26.48,26.07,24.69,24.02,23. 12,17.53,16.49,16.26,15.39,15.00,12.59.ESI-MS(m / z)604.0[M+HCOOH] - ,560.0[M+H] +.

[0050] Example 11, Synthesis of Compound 11:

[0051]

[0052] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with 2-morpholinoethane-1-amine (160 mg, 1.23 mmol) to obtain pure white solid powder 11 in 80% yield. 1 H NMR (500MHz, DMSO-d6) δ (ppm): 6.93 (t, J = 5.3Hz, 1H), 5.20 (t, J = 3.7Hz, 1H), 4.39 (t,J=5.1Hz,1H),4.14(d,J=4.9Hz,1H),3.75(s,1H),3.55(t,J=4.6Hz,5H),3.45 -3.41(m,1H),3.13-3.01(m,4H),2.54(dd,J=13.4,4.5Hz,1H),2.36(s,1H),2.33(t,J=4.7Hz,4H),2.31(s,1H),2.30(s,1H),2.28( s,1H),1.91(t,J=5.9Hz,1H),1.87–1.82(m,1H),1.61(s,1H),1.57(s,1H),1.56(d,J=2.9Hz,1H),1.53(d,J=2.9Hz,1H),1.51(s,1H ),1.49(s,1H),1.47–1.45(m,1H),1.40(d,J=4.5Hz,1H),1.37(d,J=4.1Hz,1H),1.28(s,3H),1.24(d,J=4.6Hz,1H),1.23–1.20(m,1 H),1.14(s,1H),1.12(s,1H),1.10(s,1H),1.08(s,3H),1.07(s,1H),0.86(s,3H),0.84(d,J=6.7Hz,4H),0.63(s,3H),0.53(s,3H). 13C NMR(126MHz,DMSO-d6)δ(ppm):176.74,138.85,126.81,71.95(2C),70.40, 66.24(3C),64.50,56.95,53.22(3C),52.92,46.72,46.62,46.43,41.82,41 .34,41.08,37.99,36.22(2),35.98,32.27,27.83,26.54,26.09,24.79,24. 01,23.16,17.55,16.54,16.29,15.44,12.61.ESI-MS(m / z)644.1[M+HCOOH] - ,600.2[M+H] + .

[0053] Example 12, Synthesis of Compound 12:

[0054]

[0055] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with ethane-1,2-diamine (74 mg, 1.23 mmol) to obtain pure white solid powder 12 in 80% yield. 1H NMR(500MHz,DMSO-d6)δ(ppm):7.04(t,J=5.5Hz,1H),5.20–5.18(m,1H),4.40(s, 1H),4.14(s,1H),3.45–3.42(m,2H),3.09–2.97(m,3H),2.91–2.85(m,1H),2.42( s,1H),1.93–1.86(m,1H),1.86–1.81(m,1H),1.62(d,J=4.4Hz,1H),1.60(s,1H), 1.58(d,J=3.9Hz,1H),1.56(d,J=4.0Hz,1H),1.53(d,J=3.0Hz,1H),1.51(s,1H), 1.48(d,J=3.9Hz,2H),1.47–1.45(m,2H),1.43(d,J=4.3Hz,1H),1.38(q,J=4.2,3 .5Hz,2H),1.28(s,3H),1.26(dd,J=6.6,3.2Hz,2H),1.24(d,J=3.6Hz,1H),1.21( d,J=3.2Hz,1H),1.15(d,J=3.3Hz,1H),1.12(s,1H),1.10(s,1H),1.08(s,3H),0. 86(s,3H),0.83(d,J=6.8Hz,5H),0.80(d,J=3.8Hz,1H),0.63(s,3H),0.53(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):176.87,138.88,126.73,71.92(2C),70.40,64.49,52.75,46.72,46.59,46.43,42.52,41.80,41.29,41.13,41.02 ,38.07,37.99,36.20,32.31,27.86,26.54,26.49,26.09,24.64,24.03, 23.14,17.54,16.54,16.27,15.42,12.60.ESI-MS(m / z)576.0[M+HCOOH] - ,532.1[M+H] + .

[0056] Example 13, Synthesis of Compound 13:

[0057]

[0058] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with propane-1,3-diamine (91 mg, 1.23 mmol) to obtain pure white solid powder 13 in 80% yield. 1 H NMR(500MHz,DMSO-d6)δ(ppm):7.19(t,J=5.6Hz,1H),5.19(q,J=4.6,3.6Hz,1 H),4.39(s,1H),4.14(s,1H),3.72(s,1H),3.43(dd,J=10.9,5.1Hz,2H),3.07 (d,J=10.6Hz,1H),3.03–3.00(m,1H),3.00–2.96(m,1H),2.89(d,J=6.7Hz,1H ),2.42(s,1H),1.89(dd,J=6.3,3.5Hz,1H),1.85-1.82(m,1H),1.64(d,J=4.4 Hz,1H),1.59(d,2H),1.56–1.53(m,1H),1.51(s,2H),1.48(s,2H),1.46–1.43 (m,3H),1.43–1.40(m,3H),1.38(d,J=4.9Hz,2H),1.28(s,3H),1.25–1.22(m, 2H),1.21(d,J=3.1Hz,1H),1.15(s,1H),1.12(s,1H),1.10(s,1H),1.08(s,3H ),0.86(s,3H),0.83(d,J=6.9Hz,4H),0.80(s,1H),0.62(s,3H),0.53(s,3H). 13 C NMR (126MHz, DMSO-d6) δ (ppm): 176.73, 138.84, 126.73, 71.92 (3C), 70.40, 64.50, 52.71, 46.73, 46.51, 46.44, 41.80, 41.32, 40.99, 38.12, 37.9 9,36.68,36.21(2C),32.31,32.12,27.85,26.50,26.09,24.61,24.06, 23.13,17.55,16.49,16.27,15.42,12.59.ESI-MS(m / z)590.0[M+HCOOH] - ,546.1[M+H] + .

[0059] Example 14, Synthesis of Compound 14:

[0060]

[0061] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with butane-1,4-diamine (108 mg, 1.23 mmol) to obtain pure white solid powder 14 in 85% yield. 1 H NMR(500MHz,MeOD)δ(ppm):5.35(t,J=3.6Hz,1H),3.62(d,J=4.7Hz,1H),3.60(d,J =4.5Hz,1H),3.53(d,J=10.9Hz,1H),3.29(s,1H),3.14(t,J=6.4Hz,1H),3.09–3.06 (m,1H),2.71(t,J=6.8Hz,3H),2.67(d,J=9.7Hz,1H),2.47(s,1H),1.99(s,1H),1.9 6(d,J=3.3Hz,1H),1.74(d,J=3.4Hz,1H),1.71(d,J=4.7Hz,2H),1.68(d,J=4.4Hz,2 H),1.66(d,J=3.1Hz,1H),1.65(s,1H),1.62–1.61(m,1H),1.60(s,1H),1.55(s,1H ),1.52–1.50(m,4H),1.49(s,1H),1.44(s,2H),1.40(s,1H),1.35(s,3H),1.29(s,1 H),1.26(s,1H),1.23(s,1H),1.21(s,3H),1.18(s,1H),1.16(s,1H),1.01(d,J=3.6 Hz,1H),0.97(s,3H),0.96(s,1H),0.93(d,J=6.6Hz,4H),0.75(s,3H),0.71(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):176.58,138.90,126.68,71.92(2C),70.38, 64.49,52.69,46.72,46.47,46.43,41.80(2C),41.28,41.11,40.99,38.72 ,37.98,36.20(2C),32.32,30.17,27.85,26.49,26.37,26.08,24.60,24.0 4,23.12,17.54,16.50,16.26,15.41,12.59.ESI-MS(m / z)604.0[M+HCOOH] - ,560.1[M+H] + .

[0062] Example 15, Synthesis of Compound 15:

[0063]

[0064] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with 4-methylaminopyridine (132 mg, 1.23 mmol) to obtain pure white solid powder 15 in a yield of 75%. 1 H NMR (500MHz, DMSO-d6) δ (ppm): 8.46–8.44 (m, 2H), 7.86 (t, J = 6.0Hz, 1H), 7.20 (d ,J=5.2Hz,2H),5.21–5.17(m,1H),4.40(t,J=5.1Hz,1H),4.23–4.17(m,2H),4.15 (d,J=5.0Hz,2H),3.79(s,1H),3.45-3.40(m,1H),3.07(dd,J=10.5,5.1Hz,1H),2 .58(td,J=13.4,4.5Hz,1H),2.46(s,1H),1.88(t,1H),1.68–1.64(m,2H),1.62(d ,J=2.7Hz,1H),1.60(d,J=4.3Hz,1H),1.58–1.55(m,1H),1.50(d,J=3.9Hz,2H),1 .47(t,J=4.9Hz,3H),1.44(d,J=5.2Hz,1H),1.37(d,J=12.5Hz,2H),1.29(s,3H), 1.27–1.25(m,1H),1.24(d,J=3.4Hz,1H),1.20–1.17(m,1H),1.14(d,J=3.2Hz,1H ),1.12(s,2H),1.09(s,3H),0.85(s,2H),0.84(s,4H),0.53(s,3H),0.51(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):177.08,149.22(2C),149.18,138.75,126.88,1 22.19(2C),71.92(2C),70.41,64.52,52.74,46.73,46.63,46.45,41.81,41.35 ,41.31,40.96,38.04,37.98,36.20,32.27,27.85,26.55,26.48,26.05,24.65, 24.12,23.13,17.55,16.36,16.27,15.40,12.61.ESI-MS(m / z)624.2[M+HCOOH] - ,580.1[M+H] + .

[0065] Example 16, Synthesis of Compound 16:

[0066]

[0067] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with pyridine-2-methylamine (133 mg, 1.23 mmol) to obtain pure white solid powder 16 in 80% yield. 1 H NMR (500MHz, DMSO-d6) δ (ppm): 8.43 (d, J = 2.2 Hz, 1H), 8.40 (dd, J = 4.7, 1.6 Hz, 1H), 7.79 (t, J=6.0Hz,1H),7.60(dt,J=7.9,2.1Hz,1H),7.30(dd,J=7.8,4.7Hz,1H),5.19(t,J=3.7Hz,1 H),4.38(t,J=5.1Hz,1H),4.18(t,J=5.2Hz,2H),4.14(d,J=4.9Hz,1H),3.76(s,1H),3.45- 3.40(m,2H),3.07(dd,1H),2.59–2.54(m,1H),2.45(s,1H),1.92–1.86(m,1H),1.80(dd,J= 11.1,3.6Hz,1H),1.65–1.62(m,2H),1.60(t,J=2.1Hz,1H),1.58(t,J=3.7Hz,1H),1.56–1. 54(m,1H),1.50(d,J=3.8Hz,1H),1.48(s,1H),1.47(s,2H),1.43(s,1H),1.41(d,J=4.2Hz, 1H),1.38(s,1H),1.36(s,1H),1.28(s,3H),1.24(d,J=3.0Hz,1H),1.19(s,1H),1.17(d,J= 3.1Hz,1H),1.11(s,1H),1.08(s,3H),0.84(s,2H),0.83(s,5H),0.53(s,3H),0.46(s,3H). 13C NMR(126MHz,DMSO-d6)δ(ppm):176.92,148.81,147.68,138.75,135.55,135.01, 126.85,123.17,71.92(2C),70.43,64.53,52.74,46.73,46.58,46.45,41.81,41 .31,40.96,39.98,39.24,37.99,36.20,32.27,27.83,26.56,26.48,26.04,24.6 3,24.10,23.12,17.54,16.30,16.26,15.40,12.61.ESI-MS(m / z)624.2[M+HCOOH] - ,580.1[M+H] + .

[0068] Example 17, Synthesis of Compound 17:

[0069]

[0070] The reaction and post-treatment operations were performed by replacing 3,1,4-diazacyclohexane with pyridin-2-ylmethylamine (133 mg, 1.23 mmol) to obtain pure white solid powder 17 in 85% yield. 1H NMR(500MHz,MeOD)δ(ppm):8.48(d,J=5.2Hz,1H),7.78(td,J=7.8,1.8Hz,1H),7.34(d,J=7.8Hz,1H),7.30–7.28(m,1H),5.39(t,J=3.7Hz,1H),4.40(s,2H),3.60(dd,J=11.6,4.5Hz,1H),3.52(d,J=10.8Hz,1H),2.72(td,J=13.6,4.5Hz,1H),2.48(s,1H),2.02(t,J=5.3Hz,1H),1.99(dd,J=6.9,3.7Hz,1H),1.92(dd,J=11.1,3.6Hz,1H),1.79–1.77(m,1H),1.76–1.74(m,1H),1.73(s,1H),1.72(d,J=2.4Hz,1H),1.70(s,1H),1.67(dd,J=8.2,4.0Hz,1H),1.64–1.61(m,2H),1.61–1.59(m,2H),1.57(d,J=4.1Hz,1H),1.55–1.53(m,1H),1.42(s,1H),1.38(d,J=5.7Hz,1H),1.35(s,3H),1.33(d,J=3.3Hz,1H),1.25(d,J=4.2Hz,1H),1.22(s,3H),1.20(d,J=3.0Hz,1H),1.16(d,J=2.2Hz,1H),1.14(s,1H),1.02(s,1H),0.99(d,J=4.2Hz,1H),0.97(d,J=4.7Hz,1H),0.94(d,J=6.6Hz,3H),0.90(s,3H),0.69(s,3H),0.57(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):176.98,158.84,148.48,138.69,136.37,127.04,121.79,120.96,71.93,70.39,64.49,54.85,52.93,46.71,46.61,46.42,44.30,41.79,41.30,40.97,37.97,37.89,36.16,32.20,27.82,26.53,26.48,26.07,24.78,24.07,23.11,17.51,16.27,16.14,15.37,12.57.ESI-MS(m / z)624.2[M+HCOOH] - ,580.1[M+H]+ .

[0071] Example 18, Synthesis of Compound Ast-a:

[0072]

[0073] Compound Ast was purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd. 10 g of Ast was dissolved in 200 mL of MeOH, and 12 mL of 5 N NaOH was added. The mixture was refluxed at 80°C for 4 h, then concentrated under reduced pressure. 200 mL of water was added, and 2 N HCl was slowly added dropwise with stirring at room temperature until the pH reached 3. The mixture was filtered, and the filter cake was washed with plenty of water and dried to obtain a pale yellow solid, Ast-a, with a yield of 90%. 1 HNMR (400MHz, CD3OD) δ (ppm): 5.24 (t, J = 3.6 Hz, 1H), 3.70 (ddd, J = 11.4, 9.6, 4 .5Hz,1H),3.51(d,J=11.1Hz,1H),3.36(d,J=9.6Hz,1H),3.27(d,J=11.1Hz,1 H),2.21(d,J=11.3Hz,1H),2.13–1.80(m,5H),1.77–1.23(m,11H),1.14(s,3H ),1.05(s,3H),0.97(s,3H),0.89(d,J=6.4Hz,3H),0.85(s,3H),0.70(s,3H); 13 C NMR(126MHz,MeOD)δ(ppm):181.59,139.75,126.63,78.10,69.64,66.22,54.28,48.95,48.10,47.98,44.10,43.34,40.75, 40.37,38.95,38.07,33.60,31.75,29.13,25.28,24.45,24.19,21.63,19.06,17.84,17.71,13.96; ESI-MS(m / z)488.0[MH] - ,490.1[M+H] + .

[0074] Example 19, Synthesis of Compound 18:

[0075]

[0076] Compound Ast-a (200 mg, 0.410 mmol) was dissolved in DMSO (2 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (172 mg, 0.451 mmol) and triethylamine (62.1 mg, 0.615 mmol) were added. After stirring at room temperature for 0.5 h, anhydrous piperazine (105.8 mg, 1.230 mmol) was added, and the mixture was heated and stirred at 60°C for 4 h. After completion of the reaction, the mixture was cooled to room temperature and 5 mL of water was added. A large amount of white solid precipitated. The solid was filtered, and the filter cake was washed twice with water (5 mL × 2) and ethanol (2 mL × 2), respectively, and dried to obtain 18 as a white solid powder in a yield of 79%. 1 H NMR (400MHz, CD3OD) δ (ppm): δ5.22 (t, J=3.8Hz, 1H), 3.80–3.61 (m, 5H), 3.51 ( d,J=11.2Hz,1H),3.35(d,J=9.6Hz,1H),3.26(d,J=11.2Hz,1H),2.95(br,4H), 2.40(d,J=11.1Hz,1H),2.29–2.16(m,1H),2.04–1.24(m,17H),1.14(s,3H),1. 04(s,3H),1.00–0.95(m,3H),0.91(d,J=6.4Hz,3H),0.77(s,3H),0.69(s,3H); 13 C NMR(126MHz,DMSO-d6)δ(ppm):173.93,138.85,124.22,75.55,67.44,63.90,47.54,47.06,46.95,46.47,46.05,45.79,42.50,4 1.71,38.26,37.31,33.49,32.18,30.05,27.69,23.62,23.01,21.14,17.39,16.87,16.55,13.80; ESI-MS(m / z)602.0[M+HCOOH] - ,557.9[M+H] + .

[0077] Example 20, Synthesis of Compound 19:

[0078]

[0079] The reaction and post-treatment were performed as in 18, except that piperazine was replaced by 1,4-diazacyclohexane (123.0 mg, 1.230 mmol) to afford pure white solid powder 19 in 72% yield. 1H NMR (400MHz, DMSO-d6) δ (ppm): 5.09 (s, 1H), 4.41 (t, J = 5.2Hz, 1H), 4.23 (s, 1H), 4. 26–4.12(m,1H),4.18(s,1H),3.48(s,5H),3.28(s,1H),3.17(d,J=8.6Hz,1H),3.0 3(dd,J=10.5,3.8Hz,1H),2.89–2.57(m,4H),2.37(d,J=11.2Hz,1H),2.11–1.08(m ,20H),1.04(s,3H),0.92(s,6H),0.84(d,J=6.2Hz,3H),0.69(s,3H),0.54(s,3H); 13 C NMR(101MHz,DMSO-d6)δ(ppm):174.40,138.82,124.04,75.53,67.43,63.88,47.05,46.96,46.73,46.59,46.04,45.88,45.77,42.4 8,41.89,38.32,37.31,33.70,32.22,30.15,27.85,22.99,21.19,17.51,17.37,16.88,16.71,13.80; ESI-MS(m / z)616.1[M+HCOOH] - ,572.1[M+H] + .

[0080] Example 21, Synthesis of Compound 20:

[0081]

[0082] The reaction was carried out as in 18, except that piperazine was replaced by N-methylpiperazine (123.0 mg, 1.230 mmol) to afford pure white solid powder 20 in 82% yield. 1H N R(400MHz,CD3OD)δ(ppm):5.23(t,J=3.6Hz,1H),3.89(br,4H),3.70(ddd,J=11.3,9 .6,4.5Hz,1H),3.51(d,J=11.1Hz,1H),3.36(d,J=9.6Hz,1H),3.26(d,J=11.1Hz,1H ),3.14(br,4H),2.84(s,3H),2.38(d,J=10.9Hz,1H),2.03–1.24(m,17H),1.14(s,3 H),1.04(s,3H),1.01–0.95(m,3H),0.92(d,J=6.4Hz,3H),0.76(s,3H),0.69(s,3H); 13 C NMR (126MHz, CD3OD) δ (ppm): 177.70, 139.99, 126.59, 78.13, 69.62, 66.25, 54.66, 50.03, 48.16, 47.99, 44.11, 43.87, 43.40, 40 .69,40.06,38.96,35.28,33.45,31.43,29.23,24.44,21.57,19.03,17.97,17.69,17.56,13.94; ESI-MS(m / z)615.9[M+HCOOH] - ,571.9[M+H] + .

[0083] Example 22, Synthesis of Compound 21:

[0084]

[0085] The reaction was carried out as in 18, except that piperazine was replaced by 4-dimethylaminopiperidine (175.0 mg, 1.230 mmol) to afford pure white solid powder 21 in a yield of 77%. 1H NMR (500MHz, CD3OD) δ (ppm): 5.22 (s, 1H), 4.47 (d, J = 12.6Hz, 2H), 3.69 (ddd, J = 11 .7,9.6,4.6Hz,1H),3.51(d,J=11.0Hz,1H),3.36(d,J=9.6Hz,1H),3.26(d,J=11. 0Hz,1H),2.77(s,2H),2.49–2.38(m,2H),2.33(s,6H),2.26–1.23(m,23H),1.14( s,3H),1.04(s,3H),0.98(s,2H),0.91(d,J=6.4Hz,3H),0.79(s,3H),0.70(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):177.05,140.18,126.49,78.08,69.65,66.19,63.54,48.87,48.13,47.98,44.11,43.46,41.78,40.71,40.18 ,38.97,35.27,33.58,31.54,30.01,29.43,25.05,24.46,24.21,21.66,19.04,18.07,17.73,17.69,13.96; ESI-MS(m / z)644.0[M+HCOOH] - ,600.0[M+H] + .

[0086] Example 23, Synthesis of Compound 22:

[0087]

[0088] The reaction was carried out as in 18, except that piperazine was replaced by N-methylpiperidin-4-amine (140.0 mg, 1.230 mmol) to afford pure white solid powder 22 in 80% yield. 1H NMR (400MHz, CD3OD) δ (ppm): 5.35 (t, J=3.6Hz, 1H), 3.71 (ddd, J=11.3, 9.6, 4.5Hz, 1 H),3.70–3.57(m,1H),3.52(d,J=11.1Hz,1H),3.38(d,J=9.6Hz,1H),3.28(d,J=11. 1Hz,1H),2.87(dd,J=12.4,4.5Hz,2H),2.29(s,3H),2.25–1.26(m,24H),1.16(s,3H ),1.06(s,3H),1.01–0.98(m,3H),0.93(d,J=6.5Hz,3H),0.84(s,3H),0.71(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):179.50,140.19,126.76,78.07,69.63,66.18,55.66,55.58,53.98,48.06,47.99,47.51,46.15,44.11,43.50,40.92 ,40.79,40.22,38.94,38.74,33.60,32.26,31.91,28.93,24.93,24.48, 24.06,21.60,19.01,18.27,17.70,13.97; ESI-MS(m / z)629.6[M+HCOOH] - ,586.6[M+H] + .

[0089] Example 24, Synthesis of Compound 23:

[0090]

[0091] The reaction was carried out as in step 18, except that piperazine was replaced by 4-aminopiperidine (123.0 mg, 1.230 mmol) to afford pure white solid powder 23 in a yield of 75%. 1H NMR (400MHz, CD3OD) δ (ppm): 5.33 (t, J = 3.7 Hz, 1H), 3.79 (tt, J = 11.2, 4.3 Hz, 1H), 3.69 (ddd,J=11.3,9.6,4.5Hz,1H),3.50(d,J=11.1Hz,1H),3.35(d,J=9.6Hz,1H),3.29–3.1 7(m,3H),2.83(td,J=12.6,2.9Hz,2H),2.29–2.17(m,1H),2.13–1.22(m,22H),1.15(s ,3H),1.04(s,3H),1.00–0.96(m,3H),0.92(d,J=6.5Hz,3H),0.83(s,3H),0.69(s,3H); 13 C NMR(126MHz,CD3OD+CDCl3)δ(ppm):178.85,139.57,126.06,78.48,68.93,67.1 9,53.68,48.22,48.07,47.98,47.00,46.04,44.73,44.63,43.31,43.00,40.24 ,40.14,39.54,38.43,38.12,32.98,31.33,30.82,30.36,28.29,24.60,23.95, 23.75,21.42,18.58,17.92,17.55,17.47,13.48; ESI-MS(m / z)615.9[M+HCOOH] - ,571.9[M+H] + .

[0092] Example 25, Synthesis of Compound 24:

[0093]

[0094] The reaction was carried out as in step 18, except that piperazine was replaced by 2-aminomethylpiperidine (140.0 mg, 1.230 mmol) to afford pure white solid 24 in a yield of 72%. 1H NMR (400MHz, CD3OD) δ (ppm): 5.35 (t, J = 3.7 Hz, 1H), 3.70 (ddd, J = 11.3, 9.5, 4. 5Hz,1H),3.50(d,J=11.1Hz,1H),3.36(d,J=9.6Hz,1H),3.26(d,J=11.1Hz,1H ),3.18–2.93(m,3H),2.69–2.50(m,2H),2.23–1.25(m,23H),1.15(s,3H),1.0 4(s,3H),0.99–0.96(m,3H),0.92(d,J=6.5Hz,3H),0.81(s,3H),0.69(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):180.62,140.25,126.79,78.06,69.61,66.16 ,54.09,48.07,47.97,45.38,44.93,44.13,43.43,40.89,40.78,40.26,39 .04,38.95,35.13,33.53,31.89,28.90,27.92,27.88,25.08,24.47,24.1 5,21.58,19.01,18.05,17.72,17.68,13.96; ESI-MS(m / z)630.0[M+HCOOH] - ,585.9[M+H] + .

[0095] Example 26, Synthesis of Compound 25:

[0096]

[0097] The reaction was carried out as in 18, except that piperazine was replaced by piperidin-4-ylmethylamine (140.0 mg, 1.230 mmol) to afford pure white solid powder 25 in 70% yield. 1H NMR (400MHz, CD3OD) δ (ppm): 7.48 (t, J = 6.1 Hz, 1H), 5.35 (t, J = 3.2 Hz, 1H), 3.77–3. 60(m,1H),3.50(d,J=11.0Hz,1H),3.44–3.33(m,3H),3.26(d,J=11.0Hz,1H),3.05 (m,2H),2.99–2.84(m,2H),2.19(d,J=10.4Hz,1H),2.12–1.20(m,23H),1.14(s,3H ),1.04(s,3H),0.99–0.95(m,3H),0.92(d,J=6.2Hz,3H),0.79(s,3H),0.69(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):180.63,140.26,126.80,78.07,69.62,66.17 ,54.09,48.07,47.96,45.39,44.93,44.13,43.43,40.89,40.77,40.26,39 .04,38.95,35.13,33.53,31.89,28.90,27.92,27.88,25.08,24.46,24.1 5,21.58,19.01,18.05,17.72,17.68,13.96; ESI-MS(m / z)629.9[M+HCOOH] - ,585.9[M+H] + .

[0098] Example 27, Synthesis of Compound 26:

[0099]

[0100] The reaction was carried out as in 18, except that piperazine was replaced by 2-morpholinoethane-1-amine (159.8 mg, 1.230 mmol) to afford pure white solid 26 in a yield of 75%. 1 HNMR(400MHz,CD3OD)δ(ppm):5.38(t,J=3.5Hz,1H),3.93–3.64(m,5H),3.52(d,J=11.2Hz,1H),3.37(d,J=9.5Hz,1H),3.29–3.15 (m,2H),2.49(m,6H),2.16–1.24(m,19H),1.17(s,3H),1.06(s,3H),1.00(s,3H),0.94(d,J=6.4Hz,3H),0.83(s,3H),0.71(s,3H); 13C NMR (126MHz, CD3OD) δ (ppm): 180.24, 140.32, 126.89, 78.08, 69.64, 67.87, 66.18, 58.19, 54.64, 54.39, 48.03, 44.13, 43.51, 40.88, 40.35 ,38.96,38.67,37.10,33.42,31.86,28.90,25.33,24.54,24.04,21.59,19.00,17.99,17.78,17.68,13.94; ESI-MS(m / z)645.6[M+HCOOH] - ,601.7[M+H] + .

[0101] Example 28, Synthesis of Compound 27:

[0102]

[0103] The reaction was carried out as in 18, except that piperazine was replaced by N 1 ,N 1 -dimethylethane-1,2-diamine (108.2 mg, 1.230 mmol) to give pure white solid powder 27 with a yield of 76%. 1 H NMR (400MHz, CD3OD) δ (ppm): 5.36 (t, J=3.5Hz, 1H), 3.72 (ddd, J=11.3, 9.6, 4.4Hz ,1H),3.52(d,J=11.1Hz,1H),3.38(d,J=9.6Hz,1H),3.28(d,J=11.1Hz,1H),3.19( dt,J=13.7,6.9Hz,1H),2.52–2.39(m,2H),2.31(s,6H),2.17–1.25(m,18H),1.16( s,3H),1.06(s,3H),1.00(s,3H),0.93(d,J=6.5Hz,3H),0.83(s,3H),0.71(s,3H); 13CNMR(101MHz,CD3OD)δ(ppm):180.30,140.10,126.99,78.04,69.63,66.13,58.89,54.20,48.04,47.97,45.48,44.12,43.45,40.88,40.82,4 0.32,38.93,38.62,38.02,33.45,31.88,28.89,25.25,24.50,24.08,2 1.59,19.00,17.98,17.76,17.67,13.96; ESI-MS(m / z)604.1[M+HCOOH] - ,560.0[M+H] + .

[0104] Example 29, Synthesis of Compound 28:

[0105]

[0106] The reaction was carried out as in 18, except that piperazine was replaced by N,N-dimethylpropylenediamine (108.2 mg, 1.230 mmol) to afford pure white solid powder 28 in 79% yield. 1 H NMR (400MHz, CD3OD) δ (ppm): 5.33 (t, J=3.6Hz, 1H), 3.70 (ddd, J=11.3, 9.6, 4.4Hz, 1H) ,3.50(d,J=11.1Hz,1H),3.36(d,J=9.6Hz,1H),3.26(d,J=11.1Hz,1H),3.15(ddt,J=3 5.6,13.5,6.9Hz,2H),2.34(td,J=7.1,3.2Hz,2H),2.25(s,6H),2.18–1.23(m,21H),1 .14(s,3H),1.04(s,3H),0.98(s,3H),0.92(d,J=6.5Hz,3H),0.81(s,3H),0.69(s,3H); 13 C NMR (126MHz, CD3OD) δ (ppm): 178.39, 140.07, 126.31, 78.10, 69.65, 66.22, 56.86, 50.26, 49.83, 48.15, 47.99, 45.66, 44.12, 43.60, 40.76 ,40.23,38.98,37.90,35.10,33.56,31.54,29.25,24.44,21.62,19.04,18.01,17.92,17.86,17.68,13.95; ESI-MS(m / z)617.7[M+HCOOH]- ,574.7[M+H] + .

[0107] Example 30, Synthesis of Compound 29:

[0108]

[0109] The reaction was carried out as in 18, except that piperazine was replaced by N 1 -methylethane-1,2-diamine (91.0 mg, 1.230 mmol) to give pure white solid powder 29 with a yield of 69%. 1 H NMR (400MHz, CD3OD) δ (ppm): 5.35 (t, J = 3.6Hz, 1H), 3.70 (ddd, J = 11.4, 9.5, 4.5Hz, 1H), 3.50 (d, J = 11.1Hz, 1H), 3.36 (d, 9.5Hz 1H),3.29–3.12(m,3H),2.61(t,J=6.5Hz,2H),2.37(s,3H),2.24–1.19(m,19H),1.1 5(s,3H),1.04(s,3H),0.98(s,3H),0.92(d,J=6.4Hz,3H),0.81(s,3H),0.69(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):180.45,140.22,126.83,78.06,69.62,66.16,54.03,51.51,48.90,48.05,47.97,44.12,43.43,40.86,40.80,40.27 ,39.78,38.93,38.70,35.92,33.47,31.88,28.90,25.14,24.48,24.13, 21.60,19.00,17.98,17.75,17.66,13.96; ESI-MS(m / z)590.1[M+HCOOH] - ,546.0[M+H] + .

[0110] Example 31, Synthesis of Compound 30:

[0111]

[0112] The reaction was carried out as in step 18, except that piperazine was replaced by N-methylpropylenediamine (108.2 mg, 1.230 mmol) to afford pure white solid powder 30 in a yield of 70%. 1H NMR (400MHz, CD3OD) δ (ppm): 5.34 (t, J = 3.6Hz, 1H), 3.69 (ddd, J = 11.3, 9.6, 4.5Hz, 1H), 3.50 (d, J = 11.1Hz, 1H), 3.35 (d, J = 9.6Hz, 1H), 3.28–3.08 (m, 3H),2.68(t,J=7.1Hz,2H),2.48(s,3H),2.23–1.24(m,20H),1.15(s,3H), 1.04(s,3H),0.98(s,3H),0.92(d,J=6.4Hz,3H),0.80(s,3H),0.69(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):180.79,

[0113] 140.07,126.90,78.06,69.64,66.15,54.03,48.92,48.04,47.96,44.12,43.40,40.88,40.78,40.31,38.99,38.94,3 7.91,35.09,33.45,31.87,28.87,25.10,24.47,24.14,21.58,19.00,17.99,17.74,17.65,13.94; ESI-MS(m / z)603.6

[0114] [M+HCOOH] - ,559.7[M+H] + .

[0115] Example 32, Synthesis of Compound 31:

[0116]

[0117] The reaction was carried out as in 18, except that piperazine was replaced by N 1 -ethylethane-1,2-diamine (108.2 mg, 1.230 mmol) was added to give pure white solid powder 31 with a yield of 72%. 1H NMR (400MHz, CD3OD) δ (ppm): 5.37 (t, J=3.5Hz, 1H), 3.71 (ddd, J=11.3, 9.5, 4.5Hz, 1H), 3.52 (d, J=11.0Hz, 1H), 3.44–3.35 (m, 2H), 3.31–3.22 (m, 2H), 2.89–2.73(m,4H),2.25–1.26(m,20H),1.21(t,J=7.2Hz,3H),1.17(s,3H), 1.06(s,3H),1.00(s,3H),0.94(d,J=6.5Hz,3H),0.82(s,3H),0.71(s,3H); 13 C NMR (101MHz, CD3OD) δ (ppm): 181.08, 140.03, 126.93, 78.05, 69.62, 66.15, 53.96, 48.90, 48.02, 47.94, 44.33, 44.11, 43.38, 40.86, 40.76, 40.24 ,39.06,38.92,38.67,33.39,31.83,28.86,25.12,24.44,24.13,21.58, 18.99,17.97,17.72,17.64,13.94,13.57; ESI-MS(m / z)604.1[M+HCOOH] - ,560.1[M+H] + .

[0118] Example 33, Synthesis of Compound 32:

[0119]

[0120] The reaction was carried out as in 18, except that piperazine was replaced by ethane-1,2-diamine (74.0 mg, 1.230 mmol) to afford pure white solid 32 in 50% yield. 1 H NMR (400MHz, CD3OD) δ (ppm): 5.35 (t, J = 3.6 Hz, 1H), 3.69 (ddd, J = 11.3, 9.7, 4.4 Hz, 1H), 3.50 (d, J = 11.1 Hz, 1H), 3.35 (d, J = 9.7 Hz, 1H), 3.29–3. 16(m,3H),2.79(t,J=6.4Hz,2H),2.20–1.25(m,19H),1.15(s,3H),1.0 4(s,3H),0.98(s,3H),0.92(d,J=6.5Hz,3H),0.81(s,3H),0.69(s,3H); 13C NMR(101MHz,CD3OD)δ(ppm):181.09,140.08,126.91,78.07,69.62,66.17,54.01,4 8.92,48.04,47.95,44.12,43.39,41.31,41.09,40.87,40.76,40.26,38.93,38.74 33.43 31.85,28.88,25.12,24.46,24.15,21.59,19.01,17.97,17.74,17.65,13.95; ESI-MS(m / z)576.1[M+HCOOH] - ,532.1[M+H] + .ESI-MS (m / z) 576.1 [M+HCOOH] - ,532.1[M+H] + .

[0121] Example 34, Synthesis of Compound 33:

[0122]

[0123] The reaction was carried out as in 18, except that piperazine was replaced by 1,3-propylenediamine (91.0 mg, 1.230 mmol) to afford pure white solid 33 in 44% yield. 1 H NMR (400MHz, CD3OD) δ (ppm): 5.36 (t, J = 3.6 Hz, 1H), 3.72 (ddd, J = 11.3, 9.5, 4.4 Hz, 1H), 3.52 (d, J = 11.1 Hz, 1H), 3.38 (d, J = 9.5 Hz, 1H), 3.29–3. 05(m,3H),2.66(t,J=6.9Hz,2H),2.32–1.23(m,21H),1.16(s,3H),1.0 6(s,3H),1.00(s,3H),0.93(d,J=6.5Hz,3H),0.82(s,3H),0.71(s,3H); 13C NMR(101MHz,CD3OD)δ(ppm):180.31,140.10,126.84,78.05,69.61,66.15,54.05,48.03,47.96,44.11,43.40,40.85,40.78,40.30,39.74,38 .93,38.89,37.83,33.47,32.95,31.89,28.89,25.11,24.46,24.16,21.62,19.00,17.96,17.77,17.68,13.97; ESI-MS(m / z)590.1[M+HCOOH] - ,546.1[M+H] + .

[0124] Example 35, Synthesis of Compound 34:

[0125]

[0126] The reaction was carried out as in 18, except that piperazine was replaced by butane-1,4-diamine (108.2 mg, 1.230 mmol) to afford pure white solid 34 in 44% yield. 1 H NMR (400MHz, CD3OD) δ (ppm): 5.36 (t, J=3.7Hz, 1H), 3.71 (ddd, J=11.3, 9.6, 4.5Hz, 1H) ,3.52(d,J=11.1Hz,1H),3.37(d,J=9.6Hz,1H),3.28(d,J=11.1Hz,1H),3.14(ddt,J=3 3.2,13.5,6.5Hz,2H),2.70(t,J=6.1Hz,2H),2.22–1.48(m,24H),1.45–1.26(m,3H),1 .16(s,3H),1.06(s,3H),1.00(s,3H),0.93(d,J=6.7Hz,3H),0.82(s,3H),0.71(s,3H); 13 CNMR(126MHz,CD3OD)δ(ppm):180.14,140.21,126.83,78.14,69.63,66.29,54.13,48.12,47.97,44.11,43.44,41.88,40.90,40.81,40.31,3 8.96,38.79,33.50,31.91,30.37,28.92,27.66,25.17,24.48,24.13,2 1.60,19.03,18.00,17.75,17.69,13.94; ESI-MS(m / z)604.1[M+HCOOH]- ,560.1[M+H] + .

[0127] Example 36, Synthesis of Compound 35:

[0128]

[0129] The reaction was carried out as in 18, except that piperazine was replaced by 4-methylaminopyridine (132.8 mg, 1.230 mmol) to afford pure white solid powder 35 in 85% yield. 1 H NMR (400MHz, CD3OD) δ (ppm): 8.44 (d, J = 6.1 Hz, 2H), 7.34 ( d, J = 6.1 Hz, 2H), 5.34 ( t,J=3.7Hz,1H),4.48–4.24(m,1H),3.69(ddd,J=11.3,9.6,4.4Hz,1H),3.50(d,J =11.0Hz,1H),3.35(d,J=9.6Hz,1H),3.25(d,J=11.0Hz,1H),2.27–1.20(m,21H), 1.13(s,3H),1.05–0.96(m,6H),0.92(d,J=6.2Hz,3H),0.68(s,3H),0.58(s,3H); 13 C NMR (126MHz, CD3OD) δ (ppm): 180.32, 151.14, 149.90, 139.99, 126.94, 124. 37,78.12,69.64,66.25,54.22,48.93,48.77,48.10,47.96,44.10,43.37, 43.14,40.85,40.76,40.30,38.93,33.50,31.87,28.86,25.16,24.44,24. 19,21.57,18.99,17.89,17.72,17.67,13.93; ESI-MS(m / z)623.6[M+HCOOH] - ,579.7[M+H] + .

[0130] Example 37, Synthesis of Compound 36:

[0131]

[0132] The reaction was carried out as in step 18, except that piperazine was replaced by 3-methylaminopyridine (132.8 mg, 1.230 mmol) to afford pure white solid 36 in a yield of 79%. 1H NMR (400MHz, CD3OD) δ (ppm): 8.51 (s, 1H), 8.42 (d, J = 4.6Hz, 1H), 7.80 (dt, J = 7.8, 1.9H z,1H),7.40(dd,J=7.8,4.6Hz,1H),5.33(t,J=3.5Hz,1H),4.48–4.21(m,2H),3.79–3.6 2(m,1H),3.51(d,J=11.0Hz,1H),3.36(d,J=9.4,1H),3.26(d,J=11.0Hz,1H),2.25–1. 18(m,20H),1.13(s,3H),1.00(s,6H),0.93(d,J=6.2Hz,3H),0.70(s,3H),0.48(s,3H); 13 CNMR(101MHz,CD3OD)δ(ppm):180.23,149.88,148.53,139.92,138.14,137.0 7,126.90,125.12,78.06,69.64,66.14,54.24,48.88,48.03,47.96,44.10,43 .34,41.79,40.79,40.76,40.32,38.90,33.48,31.86,28.83,25.13,24.42,2 4.13,21.56,18.96,17.81,17.70,17.65,13.94; ESI-MS(m / z)623.6[M+HCOOH] - ,579.7[M+H] + .

[0133] Example 38, Synthesis of Compound 37:

[0134]

[0135] The reaction was carried out as in step 18, except that piperazine was replaced by 2-methylaminopyridine (132.8 mg, 1.230 mmol) to afford pure white solid 37 in a yield of 78%. 1H NMR (400MHz, CD3OD) δ (ppm): 8.51 (d, J = 5.0Hz, 1H), 7.92 (t, J = 5.4Hz, 1H), 7.80 (m, 1H), 7.37 (d, J=9.0Hz,1H),7.32(dd,J=7.3,5.0Hz,1H),5.41(t,J=3.5Hz,1H),4.44(s,2H),3.70(ddd,J=11. 3,9.7,4.4Hz,1H),3.51(d,J=11.0Hz,1H),3.36(d,J=9.7Hz,1H),3.27(d,J=11.0Hz,1H),2.31– 1.22(m,21H),1.16(s,3H),1.03–0.98(m,6H),0.95(d,J=6.4Hz,3H),0.69(s,3H),0.60(s,3H); 13 C NMR(126MHz,CD3OD)δ(ppm):180.37,180.30,158.84,149.64,139.91,138.68,12 7.32,123.74,123.34,78.04,69.64,66.12,54.42,48.02,47.96,45.58,45.48,44 .11,43.38,40.86,40.83,40.34,38.89,38.64,33.39,31.89,28.87,25.38,24.4 4,24.12,21.58,18.96,17.75,17.63,17.59,13.92; ESI-MS(m / z)623.6[M+HCOOH] - ,579.7[M+H] + .

[0136] Example 39, Synthesis of Compound 38:

[0137]

[0138] Compound 2 (100 mg, 0.18 mmol) was dissolved in EA (2 mL). 1 mL of HCl-EA solution was slowly added dropwise at low temperature. The mixture was stirred for 0.5 h to precipitate a large amount of white solid. Filter and dry the solid to obtain 38 (90% yield). 1HNMR(400MHz,C5D5N)δ(ppm):5.58(s,1H),5.18(s,1H),4.40–4.15(m,2H),3.86–3.63(m,5H),3.41–3.20(m,2 H),3.12–2.86(m,4H),1.69(s,3H),1.44(s,3H),1.14(d,J=6.8Hz,3H),1.12(s,3H),1.08(s,3H),1.05(s,3H); 13 C NMR (126MHz, C5D5N) δ (ppm): 176.21, 140.65, 128.46, 74.08, 73.63, 68.46, 57.19, 50.06, 49.24, 48.39, 47.75, 47.26, 43.43, 42.82, 42.67, 40 .76,39.42,37.78,35.90,34.02,29.69,28.22,28.13,27.34,25.09,24.60,19.34,17.73,17.30,16.65,13.67; ESI-MS(m / z)602.1[M+HCOOH] - ,558.1[M+H] + .

[0139] Example 40, Synthesis of Compound 39:

[0140]

[0141] Compound 5 (100 mg, 0.17 mmol) was dissolved in EA (2 mL). 1 mL of HCl-EA solution was slowly added dropwise at low temperature. The mixture was stirred for 0.5 h to precipitate a large amount of white solid. The solid was filtered and dried to obtain 39 as a white solid in 85% yield. 1HNMR(500MHz,DMSO-d6)δ(ppm):6.77(d,J=7.8Hz,1H),5.17(d,J=3.7Hz,1H),4.39(s,1H),4.13(d,J=4.9Hz,1H),3.70(s,1H),3.54–3.47(m,1 H),3.47–3.39(m,2H),3.07(d,J=10.4Hz,1H),2.89(d,J=12.1Hz,2H),2 .44(s,2H),2.42–2.38(m,2H),1.94–1.84(m,2H),1.83-1.77(m,1H),1. 68–1.61(m,2H),1.61–1.54(m,4H),1.53(d,J=2.6Hz,1H),1.49(d,J=12.5Hz,5H),1.46(d,J=4.3Hz,1H),1.37(dd,J=12.7,5.1Hz,2H),1.28(s ,3H),1.27–1.23(m,3H),1.21-1.18(m,1H),1.13(d,J=13.1Hz,3H),1.0 8(s,3H),0.86(s,3H),0.83(d,J=6.7Hz,3H),0.65(s,3H),0.53(s,3H). 13 C NMR(126MHz,DMSO-d6)δ(ppm):176.22,139.40,127.08,72.43(2C),70.90,6 5.00,53.08,47.22,47.02,46.95,46.86,45.66,42.30(2C),41.74,41.55(2C) ),38.51,38.41,36.71,33.26,32.95,28.35,27.06,26.98,26.58,24.85,24 .48,23.65,18.05,17.24,16.77,15.95,13.11.ESI-MS(m / z)616.1[M+HCOOH] - ,572.1[M+H] + .

[0142] Example 41, Synthesis of Compound 40:

[0143]

[0144] Compound 26 (100 mg, 0.17 mmol) was dissolved in EA (2 mL). 1 mL of HCl-EA solution was slowly added at low temperature and stirred for 0.5 h. A large amount of white solid precipitated. Filtered and dried, a white solid 40 was obtained in an 88% yield.1 H NMR (400MHz, CD3OD) δ (ppm): 5.35 (t, J = 3.7 Hz, 1H), 3.70 (ddd, J = 11.3, 9.5, 4. 5Hz,1H),3.50(d,J=11.1Hz,1H),3.36(d,J=9.6Hz,1H),3.26(d,J=11.1Hz,1H ),3.18–2.93(m,3H),2.69–2.50(m,2H),2.23–1.25(m,23H),1.15(s,3H),1.0 4(s,3H),0.99–0.96(m,3H),0.92(d,J=6.5Hz,3H),0.81(s,3H),0.69(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):180.62,140.25,126.79,78.06,69.61,66.16 ,54.09,48.07,47.97,45.38,44.93,44.13,43.43,40.89,40.78,40.26,39 .04,38.95,35.13,33.53,31.89,28.90,27.92,27.88,25.08,24.47,24.1 5,21.58,19.01,18.05,17.72,17.68,13.96; ESI-MS(m / z)630.0[M+HCOOH] - ,585.9[M+H] + .

[0145] Example 42, Synthesis of Compound 41:

[0146]

[0147] Compound 27 (100 mg, 0.18 mmol) was dissolved in EA (2 mL). 1 mL of HCl-EA solution was slowly added dropwise at low temperature. The mixture was stirred for 0.5 h to precipitate a large amount of white solid. 41 was obtained by filtration and drying to obtain a white solid in a 92% yield. 1H NMR (400MHz, CD3OD) δ (ppm): 5.36 (t, J=3.5Hz, 1H), 3.72 (ddd, J=11.3, 9.6, 4.4Hz ,1H),3.52(d,J=11.1Hz,1H),3.38(d,J=9.6Hz,1H),3.28(d,J=11.1Hz,1H),3.19( dt,J=13.7,6.9Hz,1H),2.52–2.39(m,2H),2.31(s,6H),2.17–1.25(m,18H),1.16( s,3H),1.06(s,3H),1.00(s,3H),0.93(d,J=6.5Hz,3H),0.83(s,3H),0.71(s,3H); 13 C NMR(101MHz,CD3OD)δ(ppm):180.30,140.10,126.99,78.04,69.63,66.13,58.89,54.20,48.04,47.97,45.48,44.12,43.45,40.88,40.82,40 .32,38.93,38.62,38.02,33.45,31.88,28.89,25.25,24.50,24.08,21.59,19.00,17.98,17.76,17.67,13.96; ESI-MS(m / z)604.1[M+HCOOH] - ,560.0[M+H] + .

[0148] Example 43: Study confirming that GATA6 is a potential therapeutic target for cardiac hypertrophy

[0149] Experimental methods:

[0150] Cells: Extraction of primary myocardial cells from suckling rats: Disinfect 1-3 day old suckling rats by immersing in 75% alcohol. Sacrifice by cervical dislocation. Open the chest cavity with curved forceps, expose the heart, and quickly remove the heart. Place the heart in pre-cooled culture medium containing antibiotics. Before the heart stops beating, squeeze out the blood. Place the heart in another fresh culture medium and wash twice. Carefully grasp the heart with curved forceps and cut it into 6-7 equal-sized pieces. Place them in clean culture medium. Repeat the above steps to extract heart tissue from 10-15 suckling rats. Add 5 mL of digestion solution (0.1% type II collagenase); digest for 3 minutes for the first time to remove residual blood cells; digest for 5 minutes for the second time, collect the digested cells, and immediately transfer them to 5 mL of culture medium containing 10% serum, gently invert and mix, and stop digestion; digest for the third time, as in the previous step; digest for 6 minutes for the fourth time, collect the digested cells; digest for the fifth time, as in the previous step; digest for 7 minutes for the sixth time, collect the digested cells; after all myocardial tissues are digested, all collected cells are centrifuged; use 10% The cells were resuspended in FBS-containing DMEM medium and plated in two large dishes; the extracted cells were cultured in an incubator for 90 minutes; the upper suspended cells were collected by filtration through a 75μM cell sieve; the filtrate was collected and transferred to a centrifuge tube, and 5-bromodeoxyuridine (BrdU final concentration of 100μM) was added to inhibit the growth of fibroblasts; an in vitro angiotensin II (Ang II) stimulation-induced rat primary cardiomyocyte hypertrophy model was established. After culturing the extracted primary cardiomyocytes for 24 hours, they were infected with the virus solution and replaced with serum-free culture medium. Ang II was added to the culture medium at a final concentration of 1μM and the model was established for 48 hours.

[0151] In vivo AAV transfection: The diluted AAV8 viral solution was brought to room temperature and injected into the mouse via the tail vein using a mouse intravenous visual tail injection device. Gently apply pressure to stop bleeding. The control and model groups were injected with the control viral solution, while the GATA6 knockdown group was injected with the GATA6 shRNA viral solution. Subsequent experiments were conducted after stable expression was observed 3 weeks after injection.

[0152] A myocardial hypertrophy model was established by performing aortic arch stenosis (TAC) surgery in mice: mice were anesthetized with 1% sodium pentobarbital and fixed in a supine position; hair was removed with flat-head scissors, disinfected with iodine, and placed on a 37°C constant temperature heating plate; the skin was incised along the upper edge of the sternum to the fourth rib; ophthalmic scissors were used to cut the sternum along the middle of the sternum to the second rib, and the aortic arch was exposed with precision ophthalmic forceps; the aortic arch was sutured with a 27G needle using 6-0 silk thread; at this time, the pulsation of the right common carotid artery of the mouse was observed to accelerate, indicating that the operation was successful; the needle was quickly withdrawn, and the chest was sutured layer by layer; iodine disinfection was performed; routine feeding was performed for 4 weeks after the operation, and relevant indicators were tested.

[0153] Experimental results: The results are as follows Figure 1As shown, in rat primary cardiomyocytes, overexpression of GATA6 further increased the transcriptional levels of Nppa, Nppb, and Myh7, markers of cardiac hypertrophy, stimulated by angiotensin II (Ang II), while knockdown of GATA6 significantly inhibited the expression of hypertrophy-related genes. A mouse model with cardiac-specific knockdown of GATA6 was constructed by in situ injection of adeno-associated virus for in vivo validation, and a cardiac hypertrophy model was established by aortic arch stenosis (TAC) surgery in mice. The results showed that compared with the control group, the heart-to-body weight ratio, heart-to-tibial length ratio, and lung-to-body weight ratio of TAC mice were significantly increased, and knockdown of GATA6 significantly inhibited these values. In addition, knockdown of GATA6 ameliorated the cardiac dysfunction, increased expression of hypertrophy genes, and degree of fibrosis in mice induced by TAC surgery. These results indicate that GATA6 plays an important regulatory role in cardiomyocyte hypertrophy and may serve as a potential target for drug intervention.

[0154] Example 44: Study on the interaction between the compound and the target protein GATA6:

[0155] Experimental method: Insert the CM5 chip into the instrument and maintain a constant flow rate at 25°C, allowing HBS-EP buffer to continuously flow over the chip surface for about 5 minutes. The effects of the pH value, ionic strength, concentration, and flow rate of the protein solution to be immobilized on the results were examined. By analyzing the sensorgram, appropriate immobilization conditions were selected based on the pre-adsorption of the target protein. After the baseline stabilized, a 1:1 mixture of N-hydroxysuccinimide / N-ethyl-N-(dimethylaminopropyl) and carbodiimide (GE Healthcare) was injected over the chip surface to activate the carboxyl groups thereon. 200 μL of the sample was injected. A GATA6 protein solution was prepared to couple the amino groups on the protein surface with the activated carboxyl groups on the chip surface, thereby fixing the protein on the chip. A hydrochloric acid solution of ethanolamine was injected to block the residual activated carboxyl groups on the chip surface. A certain volume of HBS-EP solution was injected to wash away the non-covalently bound adsorbed conjugate and ethanolamine. After the conjugate was fixed on the chip surface, HBS-EP buffer was flowed through the system as the working solution, and the blank channel was used as the control channel to eliminate nonspecific binding as much as possible. After the baseline stabilized, different concentrations of 1-37 solution were injected to monitor the binding process in real time. Data were processed using the processing software provided by the Biacore T200 instrument, and the reaction kinetics and affinity information of binding and dissociation were obtained according to the 1:1 Langmuir binding model.

[0156] Experimental results: As shown in Table 1, many compounds have strong affinity to GATA6, and their binding affinity Kd is <1 μM.

[0157] Table 1. Affinity of compounds for GATA6

[0158]

[0159]

[0160] Example 45: Study on the anti-cardiac hypertrophy effect of compounds 1 and 5 in vitro

[0161] Experimental method: The primary experiment on suckling rats was the same as in Example 43.

[0162] Experimental results: As shown in Table 2, Ang II stimulation led to a significant increase in the transcription levels of cardiac hypertrophy marker genes Nppa, Nppb and Myh7; while 1 and 5 could inhibit the increase in the transcription levels of primary cardiomyocyte hypertrophy marker genes induced by Ang II in a concentration-dependent manner.

[0163] Table 2. Effects of Compounds 1 and 5 on the expression of hypertrophic genes in primary neonatal rat cardiomyocytes

[0164]

[0165] Example 46: Study on the effect of preventive administration of compounds 1 and 5 on improving myocardial hypertrophy

[0166] Experimental Methods: Male C57BL / 6 mice, 6-8 weeks old, weighing 18-20 g, were housed at 22 ± 2°C and 45 ± 10% humidity with free access to food and water. After acclimation for 7 days, they were used in subsequent experiments.

[0167] The method for establishing the mouse TAC model was the same as that in Example 43. Compounds 1 and 5 at a concentration of 10 mg / kg and the positive drug Enalapril at a concentration of 2 mg / kg were orally administered to the mice to investigate the improvement effects of compounds 1 and 5 on the myocardial hypertrophy model mice.

[0168] Experimental results: As shown in Table 3, compared with the Sham group, the heart-to-body weight ratio (HW / BW) and heart-to-tibial length ratio (HW / TL) values ​​of TAC mice were significantly increased, while oral administration of 1 and 5 significantly inhibited the above values ​​and the effect was better than that of the positive drug enalapril, indicating that compounds 1 and 5 can inhibit TAC-induced heart enlargement and weight gain in mice; left ventricular myocardial tissues of mice in each group were taken for RT-qPCR detection. As shown in Table 4, the transcription levels of myocardial hypertrophy marker genes in TAC group mice were significantly higher than those in Sham group, while oral administration of 1 and 5 inhibited the expression of TAC-induced myocardial hypertrophy (Nppa, Nppb, Myh7) marker gene transcription levels.

[0169] Table 3. Effects of Compounds 1 and 5 on Heart Weight in TAC Mice (n=5)

[0170]

[0171] Table 4. Effects of Compounds 1 and 5 on the Expression of Hypertrophic Genes in Cardiac Tissue of TAC Mice (n=5)

[0172]

[0173] Example 47: Study on the effect of preventive administration of compounds 1 and 5 on improving myocardial hypertrophy

[0174] Experimental method: The method for establishing the mouse TAC model was the same as that in Example 43; compounds 1, 2, and 5 were orally administered to mice at concentrations of 1 and 10 mg / kg, and the improvement effects of the compounds on the myocardial hypertrophy model mice were examined by cardiac ultrasound.

[0175] Experimental results: As shown in Table 5, compared with the Sham group, the left ventricular heart weight (LV) of TAC mice was significantly increased, and the left ventricular fraction shortening (FS) and left ventricular ejection fraction (EF) were significantly decreased, while oral administration of 1, 2, and 5 could significantly improve the cardiac dysfunction caused by TAC surgery.

[0176] Table 5. Effects of Compounds 1, 2, and 5 on Cardiac Function in TAC Mice (n=5)

[0177]

[0178]

[0179] Example 48: Study on the effect of compound 1 on improving cardiac hypertrophy in vivo

[0180] Experimental method: The method for establishing the mouse TAC model was the same as that in Example 43; after the TAC model was established, the mice were orally administered 1 at a concentration of 10 mg / kg in the 2nd and 3rd weeks, respectively, and the positive drug enalapril (Enalapril) was orally administered to the mice at a concentration of 2 mg / kg in the 2nd week to investigate the therapeutic effect of compound 1 on myocardial hypertrophy model mice.

[0181] Experimental results: As shown in Table 6, compared with TAC mice, 2 weeks and 3 weeks of treatment with compound 1 can significantly inhibit the enlargement and weight gain of the mouse heart; the left ventricular myocardial tissue of each group of mice was taken for RT-qPCR detection. As shown in Table 7, the transcription level of myocardial hypertrophy marker genes in the TAC group was significantly higher than that in the Sham group, and oral treatment with 1 can inhibit the expression of TAC-induced myocardial hypertrophy (Nppa, Nppb, Myh7) marker gene transcription levels, and the effect is better than the positive drug enalapril.

[0182] Table 6. Effects of Compound 1 on Heart Weight in TAC Mice (n=5)

[0183]

[0184] Table 7. Effects of Compound 1 on the expression of cardiac hypertrophy genes in TAC mice (n=5)

[0185]

Claims

1. Use of a pentacyclic triterpene derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing / treating myocardial hypertrophy, characterized in that: The chemical structural formula of the pentacyclic triterpene derivative is shown in formula (I): Wherein: R1 is H or OH; R2 is -CH2OH or -COOH; R3 is H or OH; R4 is 2. A pentacyclic triterpene derivative or a pharmaceutically acceptable salt thereof, characterized in that: The chemical structural formula of the pentacyclic triterpene derivative is shown in formula (I): Wherein: R1 is H; R2 is -CH2OH; R3 is OH; R4 is 3. A pentacyclic triterpene derivative or a pharmaceutically acceptable salt thereof, characterized in that: The chemical structural formula of the pentacyclic triterpene derivative is shown in formula (I): Wherein: R1 is OH; R2 is -CH2OH; R3 is H; R4 is 4. A pentacyclic triterpene derivative or a pharmaceutically acceptable salt thereof, characterized in that: The pentacyclic triterpene derivatives are selected from the following compounds:

5. A pharmaceutical composition comprising a pentacyclic triterpene derivative or a pharmaceutically acceptable salt thereof according to any one of claims 2 to 4, and at least one pharmaceutically acceptable carrier, additive or excipient.

6. Use of the pharmaceutical composition according to claim 5 in preparing a medicament for preventing / treating myocardial hypertrophy.

7. The use according to claim 6, wherein the pentacyclic triterpene derivative has a binding effect on GATA6, a potential intervention target for myocardial hypertrophy.

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