Anti-echinococcosis medicine as well as preparation method and application thereof

Abstract

The invention relates to the field of medicinal chemistry, relates to an echinococcosis-resistant medicine as well as a preparation method and application thereof, and in particular relates to a beta-carboline echinococcosis-resistant compound, a preparation method thereof, and medical application of the compound and a medicinal composition containing the compound in preparation of medicines for preventing or treating echinococcosis.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and relates to an anti-hydatid disease drug (human drug, veterinary drug) and its preparation method and application. Specifically, it relates to β-carboline alkaloid anti-hydatid disease compounds, their preparation methods, the compounds, and the medicinal compositions containing the compounds in the pharmaceutical use of preparing drugs for preventing or treating hydatid disease. Background Art

[0002] Hydatid disease, also known as echinococcosis, is a worldwide zoonosis caused by the adult or larval stages of tapeworms of the genus Echinococcus (Taeniidae). Among them, Echinococcus granulosus (Eg) and Echinococcus multilocularis (Em) are two important medical and public health species, which can cause cystic echinococcosis (CE) and alveolar echinococcosis (AE) respectively (Lancet, 2003, 362: 1295 - 1304.). The other two species, Echinococcus vogeli and Echinococcus oligarthrus, are the main causes of polycystic and unilocular hydatid diseases, but these two diseases rarely occur in humans (Amsterdam: IOS Press, 2002: 107 - 30; Acta Trop., 1997, 67: 43 - 65.). Hydatid disease is widely distributed worldwide, and there are epidemic areas almost on every continent. It not only brings a very serious disease burden, but also exerts a heavy economic pressure on each region (Acta Trop., 2003, 86: 3 - 17.).

[0003] At present, the clinical intervention means for hydatid disease mainly include three categories: surgical treatment, drug treatment, and dynamic observation. Although surgical treatment can effectively remove the lesions through surgical operations such as percutaneous aspiration of the inner capsule or complete removal of the inner capsule (Hepatogastroenterology, 1990, 37: 327 - 358.), it has significant limitations: the risk of postoperative recurrence is relatively high, and it highly depends on professional medical equipment and the operation level of surgeons (Acta Trop. 2020, 203: 105283). In this context, the importance of drug treatment has become increasingly prominent and has become an important part of the comprehensive treatment system for hydatid disease. For patients who cannot undergo surgical treatment or have postoperative recurrence, drug treatment is an indispensable core solution, and its clinical application value has been widely recognized.

[0004] Peganum harmala L. is a perennial herb of the family Zygophyllaceae and the genus Peganum. β-carboline is a kind of tricyclic alkaloid in the seeds of Peganum harmala, a plant of the family Peganaceae, with a benzindole structure.

[0005] It has been reported that the β-carboline derivative harmine (HM) has various pharmacological effects such as anti-parasitic (Tetrahedron Lett., 2010, 51(4), 583 - 585; J. Drug Target. 2004, 12(3), 165 - 175), bactericidal (Fitoterapia. 2010, 81(7), 779 - 782), anti-tumor ((Oncol. Rep. 2017, 38(5), 2927 - 2934; Phytomedicine. 2017, 28, 10 - 18; Altern. Med. 2017, 1 - 7)), and anti-depressant (Prog. Neuropsychopharmacol. Biol. Psychiatry. 2017, 79(Pt B), 258 - 267; Brain Res. Bull. 2018, 137, 294 - 300). Studies have shown that HM can effectively inhibit the growth of Echinococcus granulosus in vitro and in vivo, and can play a synergistic effect when combined with Albendazole (ABZ) (Exp. Parasitol., 2021, 226 - 227:108121.). However, HM has certain neurotoxicity, including tremors, hallucinations, excitement, and nerve inhibition, thus limiting its clinical application (Journal of Toxicology, 2021, 35(04):355 - 359.). To address the above problems, some researchers have carried out structural modification on HM. By designing and synthesizing a series of HM derivatives through structural modification of the 1st and 9th positions of β-carboline, among which DH-004 and DH-330 have obvious anti-hydatid disease efficacy (Patent No.: CN105998014B), and the median lethal concentration (LC 50 ) against Echinococcus granulosus in vitro is 47.77 ± 18.99 μM and 41.55 ± 9.48 μM respectively. However, in a mouse model infected with Echinococcus granulosus micro-cysts, the cyst inhibition rate of DH-330 is only 32.91% at a dose of 25 mg / kg (J. Biomed. Nanotechnol., 2020, 16(6):827 - 841.; DNA Cell Biol., 2021, 40(1):1 - 9.).

[0006]

[0007] On the basis of previous research, we continued to modify the structure of the β-carboline skeleton and designed and synthesized two series of 1,3-disubstituted β-carboline derivatives (Patent No.: CN202310838632). Among them, compound 1a has significant anti-hydatid disease efficacy. The LC 50 of 1a against protoscoleces (PSCs) of Echinococcus granulosus in vitro was 1.61 ± 0.12 μM, which was significantly better than that of ABZ and HM; 1a severely damaged the structural integrity of PSCs in a dose- and time-dependent manner. However, pharmacokinetic studies showed that 1a had poor stability, almost zero oral bioavailability, and very unsatisfactory metabolism after intravenous administration. However, in order to verify the in vivo activity of 1a, an intraperitoneal injection route of administration was adopted in a mouse model of hydatid disease with secondary infected microcysts. The results showed that intraperitoneal injection of 1a could effectively inhibit the growth of cysts in mice. At the same dose, the wet weight of cysts in the 1a group was lower than that in the model group, ABZ group, and HM group. In addition, at a dose of 50 mg / kg, the cyst inhibition rate of 1a was 76.7%, and there was no neurotoxicity (J. Med. Chem. 2023, 66, 24, 16680–16693.). Drug treatment for hydatid disease usually requires long-term medication. Compound 1a only has good in vivo activity when administered intraperitoneally, so it does not have the corresponding drugability. Therefore, further structural modification of β-carboline is still needed to obtain β-carboline derivatives with stronger anti-hydatid disease activity and better pharmacokinetic properties, so as to improve patient compliance and reduce the occurrence of adverse reactions. Summary of the Invention

[0008] Based on the above background, the present invention modifies different sites of β-carboline, designs and synthesizes a new class of anti-hydatid disease compounds that can effectively treat hydatid disease and have good drugability, and provides a preparation method of the above compounds, as well as pharmaceutical compositions containing these compounds, their pharmaceutically acceptable salts, and their medical uses.

[0009] Technical Solution:

[0010] In the first aspect, the present invention discloses a compound, which is a compound represented by Formula I, II, III, or IV, or a pharmaceutically acceptable salt thereof:

[0011]

[0012] Wherein, R 1 is selected from -H, -CH3, -CH2CH3, -CH2CH2CH3;

[0013] R 2 is selected from

[0014] n is 1, 2, or 3;

[0015] R3 Selected from

[0016] In formula Ⅲ The substitution position on the benzene ring is ortho or meta;

[0017] R 4 Selected from -CH3, -CH(CH3)2.

[0018] In some embodiments, the compounds represented by formula Ⅰ are selected from:

[0019] 1a-1: 4-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0020] 1a-2: N-(9-Ethyl-1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-((4-hydroxypiperidin-1-yl)methyl)benzamide;

[0021] 1a-3: 4-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-propyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0022] 1a-4: 4-(4-Hydroxypiperidin-1-yl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0023] 1a-5: N-(1-(4-Methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-(piperidin-1-yl)benzamide;

[0024] 1a-6: N-(1-(4-Methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-morpholinobenzamide;

[0025] 1a-7: N-(1-(4-Methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide;

[0026] 1a-8: 4-(Diethylamino)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0027] 1a-9: N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-(pyrrolidin-1-yl)benzamide;

[0028] 1a-10: 4-(dimethylamino)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0029] 1a-11: N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-(piperidin-1-yl)benzamide;

[0030] 1a-12: N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-morpholinobenzamide;

[0031] 1a-13: 4-(diethylamino)-N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0032] 1a-14: 4-(dimethylamino)-N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0033] 1a-15: N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide.

[0034] In some embodiments, the compounds of formula II are selected from:

[0035] 1a-16: 4-(((4-(hydroxymethyl)piperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0036] 1a-17: 4-(((4-(2-hydroxyethyl)piperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0037] 1a-18: 4-(((4-(3-hydroxypropyl)piperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0038] 1a-19: 4-((4-(2-Hydroxyethyl)piperazin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0039] 1a-20: 4-((4-(3-Hydroxypropyl)piperazin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0040] 1a-21: 4-(2-(4-Hydroxypiperidin-1-yl)ethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0041] 1a-22: 4-(3-(4-Hydroxypiperidin-1-yl)propyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0042] 1a-23: (S)-4-((2-(Hydroxymethyl)pyrrolidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0043] 1a-24: 4-((4-Hydroxy-4-methylpiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0044] 1a-25: N-(1-(4-Methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-((4-(4-nitrophenyl)piperazin-1-yl)methyl)benzamide.

[0045] In some embodiments, the compounds of formula III are selected from:

[0046] 1a-26: 2-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide;

[0047] 1a-27: 3-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide.

[0048] In some embodiments, the compounds of formula IV are selected from:

[0049] 1a-28: N-(1,9-dimethyl-9H-pyrido[3,4-b]indol-3-yl)-4-((4-hydroxypiperidin-1-yl)methyl)benzamide;

[0050] 1a-29: 4-((4-hydroxypiperidin-1-yl)methyl)-N-(9-isopropyl-1-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide.

[0051] In a second aspect, the present invention also provides a method for preparing the compound. In some embodiments, the synthetic route of compound I is as follows:

[0052]

[0053]

[0054] In some embodiments, the synthetic route of compound II is as follows:

[0055]

[0056] In some embodiments, the synthetic route of compound III is as follows:

[0057]

[0058] In some embodiments, the synthetic route of compound IV is as follows:

[0059]

[0060] In a third aspect, the present invention also provides a pharmaceutical composition, comprising a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0061] The dosage form of the pharmaceutical composition of the present invention can be prepared by those skilled in the art according to the conventional methods in the pharmaceutical field. For example, the active ingredient is mixed with one or more carriers (also called excipients), and then made into the required dosage form, including tablets, capsules, granules, aerosols; it can also be made into an intravenous injection or an intravenous injection freeze-dried preparation according to the conventional production methods of injections.

[0062] In a fourth aspect, the present invention also provides the use of the compound and the pharmaceutical composition in the preparation of a drug for preventing and / or treating hydatid disease.

[0063] Beneficial effects: The compounds of the present invention have the following excellent properties: (1) The compounds have good stability and are not easily metabolized in rat plasma and liver microsomes; (2) The raw materials for the compound synthesis process are readily available and the method is simple, and the synthesis route and synthesis method are easy to scale up; (3) Compounds 1a-1, 1a-2, and 1a-17 can effectively inhibit the growth of Echinococcus granulosus protoscoleces and destroy the structure of the parasite; (4) Compared with compound 1a (patent number: CN202310838632), compound 1a-1 has good pharmacokinetic properties in rats and high oral bioavailability; (5) In a mouse model of secondary infection with microcystic echinococcosis, compound 1a can only show a therapeutic effect by intraperitoneal injection, while compound 1a-1 can effectively inhibit cyst growth and destroy the cyst structure in vivo by gavage administration. Compound 1a-1 is more in line with the clinical actual medication situation of echinococcosis and has higher clinical application value; (6) Compound 1a-1 has good pharmacokinetic properties in sheep, a long half-life, and high oral bioavailability; (7) In a sheep model of natural infection with echinococcosis, compound 1a-1 has good pharmacokinetic properties and can effectively inhibit cyst growth and reduce cyst volume, superior to the positive drug albendazole (ABZ). Compound 1a-1 has very high veterinary application prospects; (8) Compound 1a-1 has good safety.

[0064] Verified by experiments, the compounds of the present invention also have the following excellent properties: 1a-1 was proven to be the best compound, and at a concentration of 10 μM, it could cause 91.68% death of Echinococcus granulosus protoscoleces, and it had a dose-dependence, and its in vitro insecticidal effect was equivalent to that of the lead compound 1a. Next, the results of the pharmacokinetic study in rats showed that after gavage administration of 1a-1 at a dose of 25 mg / kg, C max was 241 ng / mL, T 1 / 2 was 3.30 h, AUC 0-t was 2734 hr·ng / mL, AUC 0-∞It was 2768 hr·ng / mL, and the oral bioavailability was 27.80%. However, when 1a was administered by gavage at a dose of 50 mg / kg, the blood drug concentration of 1a was hardly detectable, so the pharmacokinetic parameters after gavage administration of 1a could not be calculated. This indicated that by introducing a methyl group at the 9-position of the β-carboline nucleus in the lead compound 1a, the oral pharmacokinetic properties of the compound 1a-1 were greatly improved, and the drug-likeness of 1a-1 was improved. Subsequently, in vivo pharmacodynamic studies were carried out on 1a-1. In a mouse model of secondary infected cystic echinococcosis, the compound 1a-1 was administered by gavage at three doses of high (50 mg / kg), medium (25 mg / kg), and low (12.5 mg / kg). After 28 days of treatment, the cyst inhibition rates were 77.67%, 70.57%, and 64.63% respectively, which were comparable to the in vivo therapeutic effects of the lead compound 1a (intraperitoneal injection at high, medium, and low doses, with cyst inhibition rates of 76.87%, 72.05%, and 67.17% respectively), and were all superior to ABZ and HM at the same dose, with significant differences. In addition, at the low dose, the therapeutic effect of the compound 1a-1 was superior to that of the high-dose groups of ABZ and HM.

[0065] To expand the application of 1a-1 in veterinary drugs, we evaluated the therapeutic effect of 1a-1 in sheep infected with Echinococcus granulosus. First, in healthy sheep, 1a-1 had good pharmacokinetic properties; the compound 1a-1 was administered by intravenous injection at a dose of 2.5 mg / kg, and C max was 251 ± 81.1 ng / mL within 24 hours, and T 1 / 2 was 12.2 hours; the compound 1a-1 was administered by gavage at a dose of 25 mg / kg, and C max was 107 ± 14.8 ng / mL within 48 hours, and the oral bioavailability was 36.9%; after the compound 1a-1 was continuously administered by gavage at a dose of 25 mg / kg for three days, C max was 324 ± 145 ng / mL within 96 hours, and T 1 / 2It was 46.5 hours and had a cumulative effect. Subsequently, we verified the therapeutic effect of 1a-1 in naturally Echinococcus-infected sheep. Compound 1a-1 could effectively inhibit the cyst growth in the infected sheep. During the treatment period, there were no abnormalities in the spirit, feeding, drinking, breathing, and activities of the sheep in the 1a-1 oral group and the 1a-1 intravenous group throughout the experimental cycle, and the body weight showed an upward trend. However, symptoms such as anorexia and diarrhea appeared in the ABZ oral group 7 days after administration; 14 days after administration in the ABZ oral group, the body weight decreased significantly, showing significant differences compared with day 0. After 14 days of treatment, autopsy found that compared with the model group, the vesicle volumes in the ABZ group, the 1a-1 oral group, and the 1a-1 intravenous group all decreased to varying degrees; among them, there was a significant statistical difference in the vesicle volume size between the 1a-1 intravenous group and the ABZ group. In addition, obvious calcification phenomena appeared in the vesicles in the 1a-1 oral group and the intravenous group. In summary, compound 1a-1 has a very significant anti-hydatid disease effect, can effectively treat hydatid disease and has good safety. Our research shows that compound 1a-1 is a very promising clinical candidate drug among the drugs for treating hydatid disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 shows the in vitro effect of the target compound against protoscoleces (PSCs) of Echinococcus granulosus at doses of 1, 5, and 10 μM.

[0067] Figure 2 is a schematic diagram of the remaining amounts of the lead compound 1a, compound 1a-1, and compound 1a-2 in rat plasma and liver microsomes after incubation at 37 °C for 120 minutes.

[0068] Figure 3 is the mean plasma concentration-time curve of the lead compound 1a (100 mg / kg), compound 1a-1 (25 mg / kg), and 1a-2 (25 mg / kg) in rats after intragastric administration.

[0069] Figure 4 is a schematic diagram of the in vivo treatment results of compound 1a-1 on mice infected with Echinococcus granulosus after 30 days of intervention.

[0070] Figure 5 is the mean plasma concentration-time curve of compound 1a-1 in sheep after intravenous injection (2.5 mg / kg), single intragastric administration (25 mg / kg), and continuous three-time intragastric administration (25 mg / kg, once a day).

[0071] Figure 6 is a schematic diagram of the in vivo treatment results of compound 1a-1 on sheep infected with hydatid disease after 14 days of intervention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0072] The following are specific implementation manners shown in the form of examples for further detailed description of the content of the present invention.

[0073] I. Synthesis of intermediates

[0074] Example 1: The synthesis step of intermediate 1-1 is to dissolve the raw material L-tryptophan (10.20 g, 50 mmol, 1.0 eq) in 80 mL of CH3COOH solution, add compound 4-methoxybenzaldehyde (55 mmol, 1.1 eq), stir at 90 °C for 3 hours, and monitor the reaction by thin layer chromatography (TLC). After the reaction is completed, adjust the reaction solution to pH = 5 - 6 with 2M NaOH solution, a large amount of yellow solid precipitates, filter by suction, and vacuum dry to obtain the yellow solid, which is intermediate 1-1.

[0075] Example 2: The synthesis step of intermediate 1-2 is to dissolve intermediate 1-1 (16.20 g, 50 mmol, 1.0 eq) in 140 mL of methanol, slowly drop thionyl chloride (17.85 g, 150 mmol, 3.0 eq) under vigorous stirring at -5 °C, heat the reaction solution to reflux for 4 hours after 0.5 hour, and monitor the reaction by TLC. After the reaction is completed, concentrate the solvent under reduced pressure, dissolve the concentrate in an appropriate amount of water, adjust the pH to 8 with 1M NaOH solution, precipitate white solid, filter by suction, and dry the filter cake under reduced pressure to obtain the white solid, which is intermediate 1-2.

[0076] Example 3: The synthesis step of intermediate 1-3 is to dissolve intermediate 1-2 (16.91 g, 50 mmol, 1.0 eq) in 150 mL of DMF solvent, add KMnO4 (11.05 g, 70 mmol, 1.4 eq) in batches under mechanical stirring at -5 °C, continue stirring for 1 hour, filter the reaction solution by suction after the reaction is completed, add the filtrate to a large amount of cold water, precipitate, and filter by suction to obtain the light brown solid, which is intermediate 1-3.

[0077] Example 4: The synthesis step of intermediate 1-5 is to dissolve intermediate 1-3 (3.32 g, 10 mmol, 1.0 eq) in 25 mL of anhydrous DMF, slowly add 60% NaH (0.36 g, 15 mmol, 1.5 eq) at -20 °C, continue stirring for 0.5 h, then slowly add intermediate 1-4 (15 mmol, 1.5 eq) to the mixed solution, and stir at room temperature for 1 h. After the reaction is completed, add 50 mL of H2O to the mixed solution, then extract with ethyl acetate, wash the organic phase with water and saturated brine, then dry with anhydrous Na2SO4, and concentrate to obtain intermediate 1-5.

[0078] Example 5: Synthesis steps of Intermediate 1-6 Dissolve Intermediate 1-5 (50 mmol, 1.0 eq) in 130 mL of methanol, then add 85% hydrazine hydrate (75.09 g, 1500 mmol, 30.0 eq) thereto, heat under reflux for 4 hours. After the reaction is completed, cool the reaction solution to 0 °C and filter by suction to obtain a light brown solid. Then add a large amount of cold water to the filtrate, and solid will continue to precipitate. Filter by suction, and the filter cake is dried under reduced pressure to obtain the product, namely Intermediate 1-6.

[0079] Example 6: Synthesis steps of Intermediate 1-7 Dissolve Intermediate 1-6 (50 mmol, 1.0 eq) in 100 mL of 2M HCl solution. Dissolve NaNO2 (10.35 g, 150 mmol, 3.0 eq) in 80 mL of H2O. Under mechanical stirring at -5 °C, slowly drop the NaNO2 solution into the 1-6 solution. After continuing to stir for 1 hour, adjust the pH of the reaction solution to 8 with 1M NaOH solution, precipitate solid, filter by suction and dry to obtain a pale yellow solid, namely Intermediate 1-7.

[0080] Example 7: Synthesis steps of Intermediate 1-8 Dissolve Intermediate 1-7 (50 mmol, 1.0 eq) in a mixed solution of 150 mL of H2O and glacial acetic acid (1:1), heat under reflux at 90 °C for 5 hours. After the reaction is completed, concentrate under reduced pressure to remove the solvent. The crude product is purified by column chromatography to obtain a pale yellow solid, namely Intermediate 1-8.

[0081] Example 8: Synthesis steps of Intermediate 1-10 Dissolve Intermediate 1-8 (2 mmol, 1.0 eq) in 5 mL of anhydrous dichloromethane, add triethylamine (0.81 g, 8 mmol, 4.0 eq). Then dissolve Intermediate 1-9 (2.4 mmol, 1.2 eq) in 3 mL of anhydrous dichloromethane and slowly drop it into the 1-7 solution under an ice bath at -5 °C. After 0.5 hour, the reaction is completed. Concentrate under reduced pressure to remove the dichloromethane solvent in the reaction solution, add 100 mL of water, extract with ethyl acetate, combine the organic phases, wash with water and saturated brine. The organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure to obtain a pale yellow solid, namely Intermediate 1-10.

[0082] Example 9: Synthesis steps of Intermediate 2-1 Refer to Example 1 for the synthesis steps of Intermediate 2-1.

[0083] Example 10: Synthesis steps of Intermediate 2-2 Refer to Example 2 for the synthesis steps of Intermediate 2-2.

[0084] Example 11: Synthesis steps of Intermediate 2-3 Refer to Example 3 for the synthesis steps of Intermediate 2-3.

[0085] Example 12: Synthesis steps of Intermediate 2-4 Refer to Example 4 for the synthesis steps of Intermediate 2-4, using raw materials 2-3 and methyl iodide.

[0086] Example 13: The synthesis steps of Intermediate 2-5 refer to Example 5, using raw material 2-4 and hydrazine hydrate.

[0087] Example 14: The synthesis steps of Intermediate 2-6 refer to Example 6, using raw material 2-5 and sodium nitrite.

[0088] Example 16: The synthesis steps of Intermediate 2-7 refer to Example 7, using raw material 2-6.

[0089] Example 17: The synthesis steps of Intermediate 2-9 are as follows: Dissolve Intermediate 2-7 (0.1 g, 0.33 mmol, 1.0 eq) in 5 mL of anhydrous DMF. During stirring, add N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (TCFH) / N-methylimidazole (NMI) or 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) / N,N-diisopropylethylamine (DIPEA) or benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU) / DIPEA, and then add Intermediate 2-8. React at room temperature for 4 hours to 24 hours. After the reaction is completed, add water to the reaction solution, extract with EA, combine the organic phases, wash the organic phases with saturated brine, dry over anhydrous Na2SO4, filter, concentrate under reduced pressure, and purify the concentrate by column chromatography to obtain Intermediate 2-9.

[0090] Example 18: The synthesis steps of Intermediate 2-10 refer to Example 8, using raw material 2-7 and 4-chloromethylbenzoyl chloride.

[0091] Example 19: The synthesis steps of Intermediate 3-1 refer to Example 1.

[0092] Example 20: The synthesis steps of Intermediate 3-2 refer to Example 2.

[0093] Example 21: The synthesis steps of Intermediate 3-3 refer to Example 3.

[0094] Example 22: The synthesis steps of Intermediate 3-4 refer to Example 4.

[0095] Example 23: The synthesis steps of Intermediate 3-5 refer to Example 5.

[0096] Example 24: The synthesis steps of Intermediate 3-6 refer to Example 6.

[0097] Example 25: The synthesis steps of Intermediate 3-7 refer to Example 7.

[0098] Example 26: The synthesis steps of Intermediate 3-9 refer to Example 8, using raw material 3-7 and Intermediate 3-8.

[0099] Example 27: The synthesis procedure of intermediate 4-1 was to disperse L-tryptophan (5 g, 24.50 mmol, 1.0 eq) in water. During stirring, 0.5 mol / L H2SO4 solution (0.012 eq) and acetaldehyde (1.30 g, 29.40 mmol, 1.2 eq) were successively added. Stirring was continued at room temperature for about 8 h, and a large amount of white insoluble matter was formed. Then, filtration, washing with water, washing with acetone, and drying were carried out to obtain a white solid, namely intermediate 4-1.

[0100] Example 28: The synthesis procedure of intermediate 4-2 referred to Example 2, using raw material 4-1 and thionyl chloride.

[0101] Example 29: The synthesis procedure of intermediate 4-3 referred to Example 3, using raw material 4-2 and potassium permanganate.

[0102] Example 30: The synthesis procedure of intermediate 4-5 referred to Example 4, using raw material 4-3 and intermediate 4-4.

[0103] Example 31: The synthesis procedure of intermediate 4-6 referred to Example 5, using raw material 4-5 and 85% hydrazine hydrate.

[0104] Example 32: The synthesis procedure of intermediate 4-7 referred to Example 6, using raw material 4-6 and sodium nitrite.

[0105] Example 33: The synthesis procedure of intermediate 4-8 referred to Example 7.

[0106] Example 34: The synthesis procedure of intermediate 4-9 referred to Example 8, using raw material 4-8 and 4-(chloromethyl)benzoyl chloride.

[0107] II. Synthesis of the target compound

[0108] Example 35: The synthesis procedure of target compound 1a-1 was to dissolve 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide (60 mg, 0.13 mmol, 1.0 eq) in acetonitrile, and add K2CO3 (53.9 mg, 0.39 mmol, 3.0 eq), KI (2.2 mg, 0.013 mmol, 0.1 eq) and 4-hydroxypiperidine (26.3 mg, 0.26 mmol, 2.0 eq). The reaction was carried out overnight under stirring at room temperature. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the crude product was purified by column chromatography to obtain 43.2 mg of the product, a pale yellow solid, with a yield of 62.89%. 11H NMR (500 MHz, DMSO-d6) δ 10.64 (s, 1H, NH), 8.82 (s, 1H, ArH), 8.29 (d, J = 7.9 Hz, 1H, ArH), 8.07 (d, J = 4.8 Hz, 2H, ArH), 7.63–7.61 (m, 3H, ArH), 7.59 (s, 1H, ArH), 7.44 (s, 2H, ArH), 7.33–7.27 (m, 1H, ArH), 7.11 (d, J = 8.5 Hz, 2H, ArH), 4.54 (s, 1H, OH), 3.86 (s, 3H, OCH3), 3.60–3.46 (m, 6H, CH, CH2, NCH3), 2.70 (s, 2H, CH2), 2.09 (s, 2H, CH2), 1.74 (s, 2H, CH2), 1.44 (s, 2H, CH2). 13 13C NMR (75 MHz, DMSO-d6) δ 165.14, 159.34, 143.40, 142.55, 141.35, 132.12, 131.27, 131.03, 130.97, 128.60, 127.79, 121.49, 120.60, 119.55, 113.26, 110.36, 104.54, 55.20, 45.49, 32.57. HRMS-ESI (m / z): [M+H] + calcd for [C 32 H 33 N4O3] + 521.25472, found 521.25413, ppm error 0.99. m.p., 219.8~220.0 °C.

[0109] Example 36: The synthesis procedure of the target compound 1a-2 was referred to Example 35. Using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-ethyl-9H-pyrido[3,4-b]indol-3-yl)benzamide as the raw material, 83.2 mg of the product was obtained as a yellow solid with a yield of 53.71%. 11H NMR(400MHz,DMSO-d6)δ10.71(s,1H,NH),8.84(s,1H,ArH),8.29(d,J=7.8Hz,1H,ArH),8.05(d,J=8.2Hz,2H,ArH),7.67–7.61(m,2H,ArH),7.59(d,J=8.7Hz,2H,ArH),7.41(d,J=8.1Hz,2H,ArH),7.29(t,J=7.8Hz,1H,ArH),7.11(d,J=8.7Hz,2H,ArH),4.57(d,J=3.7Hz,1H,OH),4.04(q,J=6.9Hz,2H,NCH2),3.86(s,3H,OCH3),3.50(s,2H,ArCH2),3.47–3.45(m,1H,CH),2.68–2.65(m,2H,CH2),2.08–2.02(m,2H,CH2),1.73–1.70(m,2H,CH2),1.43–1.36(m,2H,CH2),0.86(t,J=7.0Hz,3H,CH3). 13 13C NMR(126MHz,DMSO-d6)δ165.62,159.76,143.04,142.82 141.91,133.35,132.28,131.69,131.47,131.03,130.89,129.05,128.80,128.18,122.08,121.52,120.09,113.75,111.02,105.12,66.63,62.07,55.63,51.30,38.90,34.81,13.96.HRMS-ESI(m / z):[M+H] + calcd for[C 33 H 35 N4O3] + 535.27037,found535.27401,ppm error-2.13.m.p.,267.2~267.3℃.

[0110] Example 37: The synthesis procedure of the target compound 1a-3 was referred to Example 35, using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-n-propyl-9H-pyrido[3,4-b]indol-3-yl)benzamide as the raw material, and 60.7 mg of the product was obtained as a white solid with a yield of 53.49%. 11H NMR (400 MHz, DMSO-d6) δ 10.71 (s, 1H, NH), 8.84 (s, 1H, ArH), 8.29 (d, J = 7.8 Hz, 1H, ArH), 8.05 (d, J = 7.8 Hz, 2H, ArH), 7.68 (d, J = 8.4 Hz, 1H, ArH), 7.61 (d, J = 7.3 Hz, 1H, ArH), 7.57 (d, J = 8.4 Hz, 2H, ArH), 7.41 (d, J = 7.7 Hz, 2H, ArH), 7.28 (t, J = 7.3 Hz, 1H, ArH), 7.11 (d, J = 8.4 Hz, 2H, ArH), 4.57 (s, 1H, OH), 3.95 (t, J = 7.0 Hz, 2H, NCH2), 3.86 (s, 3H, OCH3), 3.50 (s, 2H, ArCH2), 3.45 (s, 1H, CH), 2.68–2.65 (m, 2H, CH2), 2.07–2.03 (m, 2H, CH2), 1.72–1.69 (m, 2H, CH2), 1.43–1.38 (m, 2H, CH2), 1.33–1.28 (m, 2H, CH2), 0.40 (t, J = 7.2 Hz, 3H, CH3). 13 13C NMR (75 MHz, DMSO-d6) δ 165.72, 159.81, 143.25, 142.02, 141.45, 132.08, 131.74, 131.13, 129.05, 128.97, 128.92, 128.30, 125.14, 122.13, 121.25, 120.05, 113.77, 111.27, 109.45, 105.24, 62.11, 55.77, 51.40, 45.55, 34.89, 22.24, 11.18. HRMS-ESI (m / z): [M+H] + calcd for [C 34 H 37 N4O3] + 549.28657, found 549.28473, ppm error 1.85. m.p., 245.6~245.8 °C.

[0111] Example 38: The synthesis procedure of the target compound 1a-4 was as follows: 4-(4-Acetoxypiperidin-1-yl)benzoic acid (288 mg, 1.10 mmol, 1.5 eq) was dissolved in anhydrous DMF. HATU (555 mg, 1.46 mmol, 2.0 eq) and DIPEA (377 mg, 2.92 mmol, 4.0 eq) were added, and then 1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine (220 mg, 0.73 mmol, 1.0 eq) was added. The mixture was stirred overnight at room temperature, and the reaction was monitored by TLC. After the reaction was completed, water was added to the reaction solution, and the mixture was extracted with EA. The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The concentrate was purified by column chromatography to obtain the product. Then the product was dissolved in 5 mL of methanol, and 1 M NaOH solution (87.6 mg, 2.19 mmol, 3.0 eq) was slowly added dropwise. After the reaction was completed, the solvent was removed by concentration under reduced pressure, and the residue was extracted to obtain the target compound 1a-4, a yellow solid, 60.3 mg, with a yield of 16.3%. 1 1H NMR (400 MHz, DMSO-d6) δ 10.34 (s, 1H, NH), 8.80 (s, 1H, ArH), 8.27 (d, J = 8.0 Hz, 1H, ArH), 7.99 (d, J = 8.9 Hz, 2H, ArH), 7.64–7.58 (m, 4H), 7.31–7.27 (m, 1H, ArH), 7.11 (d, J = 8.6 Hz, 2H, ArH), 6.98 (d, J = 9.0 Hz, 2H, ArH), 4.72 (d, J = 4.3 Hz, 1H, OH), 3.86 (s, 3H, OCH3), 3.73–3.66 (m, 3H, CH, CH2), 3.04–2.98 (m, 2H, CH2), 1.83–1.79 (m, 2H, CH2), 1.47–1.38 (m, 2H, CH2). 13 13C NMR (126 MHz, DMSO-d6) δ 165.41, 159.86, 153.24, 143.97, 143.54, 141.76, 132.49, 131.84, 131.69, 131.54, 129.89, 129.10, 122.89, 122.03, 121.19, 120.04, 113.87, 113.81, 110.88, 104.95, 66.44, 55.76, 45.57, 34.02, 33.12. HRMS-ESI (m / z): [M+H] + calcd for [C 31 H 31 N4O3] +507.23907, found 507.23802, ppm error 1.41.

[0112] Example 39: The synthesis step of the target compound 1a-5 is to dissolve 4-piperidinylbenzoic acid (203 mg, 1.0 mmol, 1.5 eq) in anhydrous DMF, add HBTU (502 mg, 1.32 mmol, 2.0 eq), DIPEA (28.43 mg, 0.22 mmol, 3.0 eq), and then add 1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine (200 mg, 0.66 mmol, 1.0 eq). Stir the mixture overnight at room temperature and monitor the reaction by TLC. After the reaction is completed, add water to the reaction solution, extract with EA, combine the organic phases, wash the organic phases with saturated NaCl solution, dry over anhydrous Na2SO4, filter, concentrate under reduced pressure, and purify the concentrate by column chromatography to obtain 33.0 mg of the target compound as a pale yellow solid, with a yield of 10.20%. 1 1H NMR (500 MHz, Chloroform-d) δ 9.07 (s, 1H, ArH), 8.78 (s, 1H, NH), 8.22 (d, J = 7.6 Hz, 1H, ArH), 7.88 (d, J = 8.8 Hz, 2H, ArH), 7.58–7.53 (m, 3H, ArH), 7.34 (d, J = 8.3 Hz, 1H, ArH), 7.28 (d, J = 7.5 Hz, 1H, ArH), 7.04 (d, J = 8.4 Hz, 2H, ArH), 6.90 (d, J = 8.8 Hz, 2H, ArH), 3.88 (s, 3H, OCH3), 3.42 (s, 3H, NCH3), 3.36–3.23 (m, 4H, CH2), 1.68–1.62 (m, 6H, CH2). 13 13C NMR (126 MHz, Chloroform-d) δ 165.14, 159.98, 153.97, 144.06, 143.07, 141.27, 132.77, 132.74, 131.59, 130.84, 128.76, 128.61, 123.11, 122.11, 121.69, 119.66, 114.17, 113.78, 109.65, 103.49, 55.46, 49.02, 32.79, 25.49, 24.39. HRMS-ESI (m / z): [M+H] + calcd for [C 31 H 31 N4O2] +491.24415, found 491.24328, ppm error 1.52. m.p., 124.5~125.1℃.

[0113] Example 40: The synthesis procedure of the target compound 1a-6 refers to Example 39. Using 4-morpholinobenzoic acid and 1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine as raw materials, 35.9 mg of the product was obtained as a light red solid, with a yield of 22.11%. 1 H NMR (500 MHz, DMSO-d6) δ 10.36 (s, 1H, NH), 8.80 (s, 1H, ArH), 8.27 (d, J = 7.9 Hz, 1H, ArH), 8.03 (d, J = 8.8 Hz, 2H, ArH), 7.64–7.59 (m, 4H, ArH), 7.31–7.27 (m, 1H, ArH), 7.11 (d, J = 8.6 Hz, 2H, ArH), 7.00 (d, J = 9.0 Hz, 2H, ArH), 3.86 (s, 3H, OCH3), 3.76–3.74 (m, 4H, CH2), 3.45 (s, 3H, NCH3), 3.27–3.25 (m, 4H, CH2). 13 C NMR (75 MHz, Chloroform-d) δ 164.97, 160.00, 153.56, 144.06, 142.76, 141.11, 132.80, 132.76, 131.29, 130.80, 128.77, 128.72, 124.63, 122.16, 121.57, 119.73, 114.00, 113.80, 109.63, 103.55, 66.68, 55.47, 47.91, 32.79. HRMS-ESI (m / z): [M+H] + calcd for [C 30 H 29 N4O3] + 493.22342, found 493.22260, ppm error 1.39. m.p., 247.3~247.5℃.

[0114] Example 41: The synthesis procedure of the target compound 1a-7 was as follows: 4-Piperazinylbenzoic acid (86 mg, 0.39 mmol, 1.5 eq) was dissolved in anhydrous DMF, and HATU (198 mg, 0.52 mmol, 2.0 eq) and DIPEA (100 mg, 0.78 mmol, 3.0 eq) were added. Then, 1-(4-Methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine (80 mg, 0.26 mmol, 1.0 eq) was added, and the mixture was stirred overnight at room temperature while monitoring the reaction by TLC. After the reaction was completed, water was added to the reaction solution, and the mixture was extracted with EA. The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 57.2 mg of the product as a light brown solid with a yield of 42.90%. 1 H NMR(500MHz,Chloroform-d)δ9.05(s,1H,ArH),8.65(s,1H,NH),8.23(d,J=7.7Hz,1H,ArH),7.90(d,J=9.1Hz,2H,ArH),7.60–7.55(m,3H,ArH),7.38(d,J=8.2Hz,1H,ArH),7.29(t,J=7.5Hz,1H,ArH),7.07(d,J=8.7Hz,2H,ArH),6.94(d,J=8.9Hz,2H,ArH),3.91(s,3H,OCH3),3.46(s,3H,N CH3),3.40–3.38(m,4H,CH2),2.64–2.62(m,4H,CH2),2.40(s,3H,CH2N CH3 ). 13 C NMR(126MHz,Chloroform-d)δ165.02,160.02,153.38,144.08,142.89,141.28,132.85,132.76,131.58,130.82,128.75,128.65,124.29,122.14,121.68,119.71,114.29,113.83,109.64,103.54,55.48,54.78,47.56,46.01,32.79.HRMS-ESI(m / z):[M+H] + calcd for[C 31 H 32 N5O2] + 506.25505,found 506.25415,ppm error1.55.m.p.,194.4~194.6℃.

[0115] Example 42: The synthesis procedure of the target compound 1a-8 was as follows: 4-Diethylaminobenzoic acid (77 mg, 0.40 mmol, 1.2 eq) was dissolved in anhydrous DMF, then TCFH (134 mg, 0.48 mmol, 1.44 eq) and NMI (136 mg, 1.65 mmol, 5.0 eq) were added. Subsequently, 1-(4-Methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine (100 mg, 0.33 mmol, 1.0 eq) was added, and the mixture was stirred overnight at room temperature while monitoring the reaction by TLC. After the reaction was completed, water was added to the reaction mixture, and the mixture was extracted with EA. The organic phases were combined, washed with saturated NaCl solution, dried over anhydrous Na2SO4, filtered, concentrated under reduced pressure, and the concentrate was purified by column chromatography to obtain 110.0 mg of the product as a pale yellow solid with a yield of 69.73%. 1 H NMR(500MHz,Chloroform-d)δ9.08(s,1H,ArH),8.69(s,1H,NH),8.23(d,J=7.7Hz,1H,ArH),7.88(d,J=8.9Hz,2H,ArH),7.59–7.56(m,3H,ArH),7.36(d,J=8.3Hz,1H,ArH),7.28(d,J=7.2Hz,1H,ArH),7.06(d,J=8.5Hz,2H,ArH),6.68(d,J=9.0Hz,2H,ArH),3.90(s,3H,OCH3),3.44(s,3H,NCH3),3.41(t,J=7.1Hz,4H,CH2),1.21(t,J=7.1Hz,6H,CH3). 13 C NMR(126MHz,Chloroform-d)δ165.32,160.02,150.40,144.13,143.23,141.11,132.86,132.74,130.86,129.12,128.60,124.42,122.19,121.74,120.32,119.65,113.83,110.61,109.60,103.47,55.49,44.50,32.79,12.58.HRMS-ESI(m / z):[M+H] + calcd for[C 30 H 31 N4O2] + 479.24415,found 479.24350,ppm error 1.03.m.p.,207.9~208.1℃.

[0116] Example 43: The synthesis procedure of the target compound 1a-9 was referred to Example 41. Using 4-pyrrolidinylbenzoic acid and 1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine as raw materials, 18.2 mg of the product was obtained, which was a yellow solid with a yield of 23.86%. 1 H NMR(500MHz,Chloroform-d)δ9.08(s,1H,ArH),8.70(s,1H,NH),8.23(d,J=7.7Hz,1H,ArH),7.88(d,J=8.6Hz,2H,ArH),7.57–7.54(m,3H,ArH),7.33(d,J=8.3Hz,1H,ArH),7.27(t,J=7.7Hz,1H,ArH),7.05(d,J=8.2Hz,2H,ArH),6.53(d,J=8.6Hz,2H,ArH),3.88(s,3H,OCH3),3.41(s,3H,NCH3),3.32–3.30(m,4H,CH2),2.00(s,4H,CH2). 13 C NMR(126MHz,Chloroform-d)δ165.44,159.98,150.25,144.06,143.25,141.22,132.75,132.71,131.64,130.86,128.91,128.58,122.08,121.71,120.45,119.61,113.79,111.01,109.65,103.39,55.48,47.57,32.80,25.51.HRMS-ESI(m / z):[M+H] + calcd for[C 30 H 29 N4O2] + 477.22850,found 477.22787,ppm error 1.13.m.p.,245.8~246.2℃.

[0117] Example 44: The synthesis procedure of the target compound 1a-10 was referred to Example 42. Using 4-dimethylaminobenzoic acid and 1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-amine as raw materials, 22.0 mg of the product was obtained, which was a yellow solid with a yield of 14.81%. 11H NMR (500 MHz, Chloroform-d) δ 9.07 (s, 1H, ArH), 8.68 (s, 1H, NH), 8.23 (d, J = 7.7 Hz, 1H, ArH), 7.90 (d, J = 9.0 Hz, 2H, ArH), 7.59–7.56 (m, 3H, ArH), 7.36 (d, J = 8.3 Hz, 1H, ArH), 7.28 (t, J = 7.6 Hz, 1H, ArH), 7.06 (d, J = 8.7 Hz, 2H, ArH), 6.71 (d, J = 8.9 Hz, 2H, ArH), 3.90 (s, 3H, OCH3), 3.44 (s, 3H, NCH3), 3.04 (s, 6H, CH3). 13 13C NMR (126 MHz, Chloroform-d) δ 165.34, 160.01, 152.75, 144.10, 143.13, 141.19, 132.81, 132.78, 131.63, 130.84, 128.81, 128.61, 122.17, 121.73, 121.28, 119.66, 113.83, 111.18, 109.61, 103.47, 55.49, 40.14, 32.79. HRMS-ESI (m / z): [M+H] + calcd for [C 28 H 27 N4O2] + 451.21285, found 451.21271, ppm error 0.52. m.p., 193.8~194.1 °C.

[0118] Example 46: The synthesis procedure of the target compound 1a-11 was referred to Example 42, using 4-piperidinylbenzoic acid and 1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-amine as raw materials, to obtain 37.8 mg of the product, a yellow solid, with a yield of 22.95%. 11H NMR (500 MHz, Chloroform-d) δ 8.99 (s, 1H, ArH), 8.70 (s, 1H, NH), 8.39 (s, 1H, NH), 8.15 (d, J = 7.8 Hz, 1H, ArH), 7.93–7.90 (m, 4H, ArH), 7.52 (ddd, J = 8.1, 6.9, 1.2 Hz, 1H, ArH), 7.47 (d, J = 8.0 Hz, 1H, ArH), 7.27 (t, J = 7.1 Hz, 1H, ArH), 7.11 (d, J = 8.7 Hz, 2H, ArH), 6.95 (d, J = 9.0 Hz, 2H, ArH), 3.89 (s, 3H, OCH3), 3.34 (t, J = 4.9 Hz, 4H, CH2), 1.73–1.69 (m, 4H, CH2), 1.67–1.63 (m, 2H, CH2). 13 13C NMR (126 MHz, Chloroform-d) δ 165.24, 160.25, 154.02, 143.95, 141.63, 132.58, 129.41, 128.78, 128.62, 123.07, 122.43, 122.34, 120.14, 114.68, 114.21, 113.47, 111.54, 55.45, 49.04, 25.49, 24.40. HRMS-ESI (m / z): [M+H] + calcd for [C 30 H 29 N4O2] + 477.22850, found 477.22747, ppm error 1.87. m.p., 159.4~159.7 °C.

[0119] Example 47: The synthesis procedure of the target compound 1a-12 was referred to Example 42. Using 4-morpholinobenzoic acid and 1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-amine as raw materials, 57.4 mg of the product was obtained, which was a yellow solid with a yield of 34.71%. 11H NMR(500 MHz, Chloroform-d) δ 8.94 (s, 1H, ArH), 8.84 (s, 1H, NH), 8.63 (s, 1H, NH), 8.10 (d, J = 7.8 Hz, 1H, ArH), 7.89 (dd, J = 17.1, 8.8 Hz, 4H, ArH), 7.52–7.46 (m, 2H, ArH), 7.24 (d, J = 7.1 Hz, 1H, ArH), 7.05 (d, J = 8.6 Hz, 2H, ArH), 6.90 (d, J = 8.8 Hz, 2H, ArH), 3.85 (d, J = 4.8 Hz, 4H, CH2), 3.84 (s, 3H, CH3), 3.26 (d, J = 4.5 Hz, 4H, CH2). 13 13C NMR(126 MHz, Chloroform-d) δ 165.13, 160.31, 153.63, 143.72, 141.70, 132.64, 130.84, 129.39, 128.79, 128.75, 124.59, 122.32, 120.32, 120.20, 114.71, 114.04, 111.59, 103.46, 66.68, 55.46, 47.95. HRMS-ESI (m / z): [M+H] + calcd for [C 29 H 27 N4O3] + 479.20777, found 479.20682, ppm error 1.62. m.p., 173.1~173.3 °C.

[0120] Example 48: The synthesis procedure of the target compound 1a-13 was referred to Example 42, using 4-diethylaminobenzoic acid and 1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-amine as raw materials, and 37.5 mg of the product was obtained as a yellow solid with a yield of 23.36%. 11H NMR(500 MHz, Chloroform-d) δ 8.98 (s, 1H, ArH), 8.74 (s, 1H, NH), 8.56 (s, 1H, NH), 8.11 (d, J = 7.7 Hz, 1H, ArH), 7.92–7.86 (m, 4H, ArH), 7.52–7.46 (m, 2H, ArH), 7.24 (t, J = 7.1 Hz, 1H, ArH), 7.06 (d, J = 8.2 Hz, 2H, ArH), 6.67 (d, J = 8.6 Hz, 2H, ArH), 3.84 (s, 3H, OCH3), 3.42 (q, J = 6.8 Hz, 4H, CH2), 1.21 (t, J = 7.1 Hz, 6H, CH3). 13 13C NMR(126 MHz, Chloroform-d) δ 165.40, 160.20, 150.40, 144.18, 141.63, 132.55, 132.25, 130.82, 129.41, 129.11, 128.55, 122.47, 122.33, 120.29, 120.07, 114.65, 111.51, 110.63, 110.21, 103.42, 55.44, 44.51, 12.58. HRMS-ESI (m / z): [M+H] + calcd for [C 29 H 29 N4O2] + 465.22850, found 465.22762, ppm error 1.61. m.p., 131.0~131.2 °C.

[0121] Example 49: The synthesis procedure of the target compound 1a-14 was referred to Example 42, using 4-dimethylaminobenzoic acid and 1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-amine as raw materials, and 23.6 mg of the product was obtained as a yellow solid with a yield of 14.45%. 11H NMR(500 MHz, Methanol-d4) δ 8.99 (s, 1H, ArH), 8.68 (s, 1H, NH), 8.46 (s, 1H, NH), 8.14 (d, J = 7.8 Hz, 1H, ArH), 7.91–7.88 (m, 4H, ArH), 7.51 (td, J = 7.0, 1.0 Hz, 1H, ArH), 7.46 (d, J = 8.1 Hz, 1H, ArH), 7.25 (td, J = 6.9, 1.1 Hz, 1H, ArH), 7.08 (d, J = 8.7 Hz, 2H, ArH), 6.72 (d, J = 9.0 Hz, 2H, ArH), 3.86 (s, 3H, OCH3), 3.05 (s, 6H, NCH3). 13 13C NMR(126 MHz, Chloroform-d) δ 165.41, 160.26, 152.78, 144.09, 141.61, 132.60, 130.80, 129.39, 128.82, 128.61, 122.47, 122.37, 121.22, 120.14, 114.72, 111.50, 111.21, 103.40, 55.48, 40.14. HRMS-ESI (m / z): [M+H] + calcd for [C 27 H 25 N4O2] + 437.19720, found 437.19626, ppm error 1.85. m.p., 160.6~160.8 °C.

[0122] Example 50: The synthesis procedure of the target compound 1a-15 was referred to Example 42. Using 4-piperazinylbenzoic acid and 1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-amine as raw materials, 46.0 mg of the product was obtained, which was a light brown solid with a yield of 26.97%. 11H NMR(500MHz, Methanol-d4) δ 8.78 (s, 1H, ArH), 8.17 (d, J = 8.1 Hz, 1H, ArH), 7.96 (t, J = 8.1 Hz, 4H, ArH), 7.62 (d, J = 8.2 Hz, 1H, ArH), 7.56 (d, J = 7.0 Hz, 1H, ArH), 7.28 (d, J = 6.8 Hz, 1H, ArH), 7.17 (d, J = 8.5 Hz, 2H, ArH), 7.02 (d, J = 8.5 Hz, 2H, ArH), 3.93 (s, 3H, OCH3), 2.66 (d, J = 4.8 Hz, 4H, CH2), 2.42 (s, 3H, NCH3), 1.35–1.32 (m, 4H, CH2). 13 13C NMR(126MHz, Methanol-d4) δ 166.09, 160.34, 153.34, 142.80, 142.48, 140.32, 132.07, 130.90, 130.17, 129.47, 128.61, 128.19, 123.72, 121.56, 121.22, 119.27, 113.93, 113.88, 111.83, 103.80, 54.49, 54.17, 46.68, 44.52. HRMS-ESI (m / z): [M+H] + calcd for [C 30 H 30 N5O2] + 492.23940, found 492.23821, ppm error 2.24. m.p., 148.3~148.6 °C.

[0123] Example 51: The synthesis procedure of the target compound 1a-16 was referred to Example 35. Using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxymethylpiperidine as raw materials, 30.0 mg of the product was obtained, which was a pale yellow solid with a yield of 52.49%. 11H NMR (500 MHz, DMSO-d6) δ 10.63 (s, 1H, NH), 8.82 (s, 1H, ArH), 8.29 (d, J = 7.8 Hz, 1H, ArH), 8.05 (d, J = 7.2 Hz, 2H, ArH), 7.68–7.59 (m, 4H, ArH), 7.42 (d, J = 7.7 Hz, 2H, ArH), 7.30 (s, 1H, ArH), 7.11 (d, J = 8.1 Hz, 2H, ArH), 4.38 (s, 1H, OH), 3.86 (s, 3H, OCH3), 3.52 (s, 2H, ArCH2), 3.46 (s, 3H, NCH3), 3.26–3.25 (m, 2H, CH2), 2.82–2.80 (m, 2H, CH2), 1.95–1.91 (m, 2H, CH2), 1.64–1.62 (m, 2H, CH2), 1.36–1.33 (m, 1H, CH), 1.23–1.12 (m, 2H, CH2). 13 13C NMR (126 MHz, DMSO-d6) δ 165.76, 159.90, 143.97, 143.30, 143.13, 141.91, 133.51, 132.67, 131.82, 131.61, 131.55, 129.17, 128.97, 128.31, 122.07, 121.16, 120.12, 113.83, 110.93, 105.12, 66.47, 62.60, 55.76, 53.69, 38.85, 33.15, 29.28. HRMS-ESI (m / z): [M+H] + calcd for [C 33 H 35 N4O3] + 535.27037, found 535.26850, ppm error 3.33. m.p., 247.5~247.7 °C.

[0124] Example 52: The synthesis procedure of the target compound 1a-17 was referred to Example 35. Using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-piperidinoethanol as raw materials, 40.6 mg of the product was obtained as a white solid, with a yield of 67.41%. 11H NMR (300 MHz, Chloroform-d) δ 9.06 (s, 1H, ArH), 8.79 (s, 1H, NH), 8.24 (d, J = 7.8 Hz, 1H, ArH), 7.95 (d, J = 8.2 Hz, 2H, ArH), 7.61 (d, J = 7.2 Hz, 1H, ArH), 7.56 (d, J = 8.7 Hz, 2H, ArH), 7.50 (d, J = 8.2 Hz, 2H, ArH), 7.39 (d, J = 8.3 Hz, 1H, ArH), 7.30 (t, J = 7.5 Hz, 1H, ArH), 7.07 (d, J = 8.7 Hz, 2H, ArH), 3.91 (s, 3H, OCH3), 3.69 (t, J = 6.4 Hz, 2H, OCH2), 3.63 (s, 2H, ArCH2), 3.47 (s, 3H, NCH3), 2.94 (d, J = 11.3 Hz, 2H, CH2), 2.07 (t, J = 10.9 Hz, 2H, CH2), 1.71 (d, J = 11.9 Hz, 2H, CH2), 1.54–1.50 (m, 2H, CH2), 1.37–1.36 (m, 3H, CH, CH2). 13 13C NMR (126 MHz, Chloroform-d) δ 165.26, 160.05, 144.08, 142.84, 142.59, 141.40, 133.46, 132.97, 132.75, 131.51, 130.81, 129.39, 128.73, 127.17, 122.17, 121.64, 119.79, 113.83, 109.68, 103.69, 63.00, 60.45, 55.49, 53.93, 39.41, 32.82, 32.29, 26.97. HRMS-ESI (m / z): [M+H] + calcd for [C 34 H 37 N4O3] + 549.28602, found 549.28485, ppm error 2.08. m.p., 243.6~243.7 °C.

[0125] Example 53: The synthetic procedure of the target compound 1a-18 was referred to Example 35, using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-(3-hydroxypropyl)piperidine as raw materials to obtain 20.0 mg of the product as a white solid with a yield of 56.42%. 11H NMR (400 MHz, Chloroform-d) δ 9.06 (s, 1H, ArH), 8.75 (s, 1H, NH), 8.25 (d, J = 7.8 Hz, 1H, ArH), 7.93 (d, J = 8.2 Hz, 2H, ArH), 7.61 (d, J = 7.3 Hz, 1H, ArH), 7.56 (d, J = 8.7 Hz, 2H, ArH), 7.47 (d, J = 8.0 Hz, 2H, ArH), 7.39 (d, J = 8.3 Hz, 1H, ArH), 7.30 (t, J = 7.4 Hz, 1H, ArH), 7.07 (d, J = 8.7 Hz, 2H, ArH), 3.91 (s, 3H, OCH3), 3.63 (t, J = 6.6 Hz, 2H, CH2O), 3.58 (s, 2H, ArCH2), 3.47 (s, 3H, NCH3), 2.89 (d, J = 10.8 Hz, 2H, CH2), 2.00 (t, J = 8.8 Hz, 2H, CH2), 1.68 (d, J = 10.0 Hz, 2H, CH2), 1.59–1.56 (m, 2H, CH2), 1.33–1.28 (m, 5H, CH, CH2). 13 13C NMR (126 MHz, Chloroform-d) δ 165.27, 160.04, 144.07, 142.94, 142.62, 141.37, 133.39, 132.95, 132.75, 131.48, 130.81, 129.38, 128.73, 127.16, 122.15, 121.63, 119.79, 113.81, 109.69, 103.69, 63.05, 55.48, 54.02, 35.58, 32.82, 32.65, 32.36, 30.09, 29.75. HRMS-ESI (m / z): [M+H] + calcd for [C 35 H 39 N4O3] + 563.30167, found 563.30135, ppm error 0.41. m.p., 85.0~85.3 °C.

[0126] Example 54: The synthesis procedure of the target compound 1a-19 was referred to Example 35. Using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and N-hydroxyethylpiperazine as raw materials, 33.2 mg of the product was obtained as a white solid with a yield of 59.66%. 11H NMR (300 MHz, Chloroform-d) δ 9.06 (s, 1H, ArH), 8.90 (s, 1H, NH), 8.24 (d, J = 7.7 Hz, 1H, ArH), 7.93 (d, J = 8.2 Hz, 2H, ArH), 7.59 (t, J = 8.2 Hz, 1H, ArH), 7.54 (d, J = 8.7 Hz, 2H, ArH), 7.43 (d, J = 8.2 Hz, 2H, ArH), 7.38 (d, J = 8.3 Hz, 1H, ArH), 7.29 (t, J = 7.7 Hz, 1H, ArH), 7.05 (d, J = 8.7 Hz, 2H, ArH), 3.90 (s, 3H, OCH3), 3.61 (t, J = 5.4 Hz, 2H, OCH2), 3.55 (s, 2H, ArCH2), 3.45 (s, 3H, NCH3), 2.84 (s, 2H, CH2), 2.57–2.49 (m, 8H, CH2). 13 13C NMR (126 MHz, Chloroform-d) δ 165.23, 160.05, 144.07, 142.55, 142.52, 141.44, 133.66, 133.00, 132.73, 131.50, 130.80, 129.29, 128.73, 127.24, 122.16, 121.63, 119.80, 113.83, 109.69, 103.69, 62.53, 59.38, 57.76, 55.49, 53.11, 52.92, 32.81. HRMS-ESI (m / z): [M+H] + calcd for [C 33 H 36 N5O3] + 550.28127, found 550.28027, ppm error 1.45. m.p., 101.8~102.1 °C.

[0127] Example 54: The synthesis procedure of the target compound 1a-20 was referred to Example 35, using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 1-piperazinylpropanol as raw materials to obtain 25.2 mg of the product as a white solid, with a yield of 40.72%. 11H NMR(300MHz,Chloroform-d)δ9.06(s,1H,ArH),8.73(s,1H,NH),8.24(d,J=7.7Hz,1H,ArH),7.93(d,J=8.2Hz,2H,ArH),7.61(t,J=8.1Hz,2H,ArH),7.56(d,J=8.7Hz,2H,ArH),7.45(d,J=8.2Hz,2H,ArH),7.39(d,J=8.4Hz,1H,ArH),7.30(t,J=7.5Hz,1H,ArH),7.07(d,J=8.7Hz,2H,ArH),3.91(s,3H,OCH3),3.83(t,J=5.1Hz,2H,CH2O),3.60(s,2H,ArCH2),3.47(s,3H,NCH3),2.83–2.80(m,4H,CH2),2.67–2.65(m,4H,CH2),1.83(p,J=5.4Hz,2H,CH2),1.30–1.23(m,2H,CH2). 13 13C NMR(126MHz,Chloroform-d)δ165.17,160.05,144.07,142.51,142.41,141.46,133.72,133.02,132.71,131.52,130.80,129.28,128.72,127.24,122.15,121.64,119.79,113.84,109.69,103.65,64.12,62.37,58.57,55.49,53.27,52.82,32.82,26.97.HRMS-ESI(m / z):[M+H] + calcd for[C 34 H 38 N5O3] + 564.29692,found564.29659,ppm error 0.36.m.p.,100.1~100.3℃.

[0128] Example 55: The synthesis procedure of the target compound 1a-21 was referred to Example 35, using 4-(2-bromoethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxypiperidine as raw materials, and 30.6 mg of the product was obtained as a white solid with a yield of 33.10%. 11H NMR (500 MHz, DMSO-d6) δ 10.60 (s, 1H, NH), 8.82 (s, 1H, ArH), 8.28 (d, J = 7.8 Hz, 1H, ArH), 8.02 (d, J = 8.1 Hz, 2H, ArH), 7.64–7.58 (m, 3H, ArH), 7.58 (s, 1H, ArH), 7.34 (d, J = 8.1 Hz, 2H, ArH), 7.29 (t, J = 6.0 Hz, 1H, ArH), 7.10 (d, J = 8.6 Hz, 2H, ArH), 4.54 (s, 1H, OH), 3.86 (s, 3H, OCH3), 3.45 (s, 4H, NCH3, CH), 2.80 (t, J = 7.2 Hz, 4H, ArCH2, CH2), 2.56–2.55 (m, 2H, CH2), 2.14–2.06 (m, 2H, CH2), 1.73–1.71 (m, 2H, CH2), 1.40–1.36 (m, 2H, CH2). 13 13C NMR (126 MHz, DMSO-d6) δ 165.76, 159.89, 145.06, 143.96, 143.16, 141.89, 132.65, 132.57, 131.82, 131.61, 131.54, 129.16, 129.06, 128.38, 122.06, 121.16, 120.11, 113.82, 110.92, 105.12, 59.68, 55.76, 51.37, 41.92, 34.84, 33.13, 11.67. HRMS-ESI (m / z): [M+H] + calcd for [C 33 H 35 N4O3] + 535.27037, found 535.26975, ppm error 0.89. m.p., 217.9~218.0 °C.

[0129] Example 56: The synthesis procedure of the target compound 1a-22 was referred to Example 35. Using 4-(2-bromopropyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxypiperidine as raw materials, 39.6 mg of the product was obtained as a white solid, with a yield of 24.85%. 11H NMR (400 MHz, Chloroform-d) δ 9.06 (s, 1H, NH), 8.76 (s, 1H, ArH), 8.24 (d, J = 7.8 Hz, 1H, ArH), 7.91 (d, J = 8.2 Hz, 2H, ArH), 7.60 (t, J = 8.0 Hz, 1H, ArH), 7.56 (d, J = 8.7 Hz, 2H, ArH), 7.39 (d, J = 8.3 Hz, 1H, ArH), 7.30 (t, J = 8.7 Hz, 3H, ArH), 7.07 (d, J = 8.7 Hz, 2H, ArH), 3.91 (s, 3H, OCH3), 3.78 (s, 1H, CH), 3.46 (s, 3H, NCH3), 2.84 (s, 2H, CH2), 2.72 (t, J = 7.6 Hz, 2H, ArCH2), 2.48–2.46 (m, 2H, CH2), 2.02–1.94 (m, 4H, CH2), 1.69–1.62 (m, 4H, CH2). 13 13C NMR (75 MHz, Chloroform-d) δ 165.26, 159.99, 146.29, 144.00, 142.51, 141.35, 132.92, 132.67, 132.38, 131.47, 130.77, 128.78, 128.69, 127.31, 122.14, 121.57, 119.74, 113.78, 109.65, 103.61, 57.57, 55.47, 53.45, 50.78, 33.98, 33.52, 32.79, 28.16. HRMS-ESI (m / z): [M+H] + calcd for [C 34 H 37 N4O3] + 549.28602, found 549.28487, ppm error 1.73. m.p., 222.1~222.3 °C.

[0130] Example 57: The synthesis procedure of the target compound 1a-23 was referred to Example 35. Using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and L-prolinol as raw materials, 35.6 mg of the product was obtained, which was a pale yellow solid with a yield of 61.00%. 11H NMR (400 MHz, Chloroform-d) δ 9.06 (s, 1H, ArH), 8.75 (s, 1H, NH), 8.25 (d, J = 7.8 Hz, 1H, ArH), 7.95 (d, J = 8.1 Hz, 2H, ArH), 7.61 (t, J = 7.4 Hz, 1H, ArH), 7.56 (d, J = 8.7 Hz, 2H, ArH), 7.46 (d, J = 7.9 Hz, 2H, ArH), 7.39 (d, J = 8.3 Hz, 1H, ArH), 7.30 (t, J = 7.5 Hz, 1H, ArH), 7.07 (d, J = 8.6 Hz, 2H, ArH), 4.08 (d, J = 13.3 Hz, 1H, OH), 3.91 (s, 3H, OCH3), 3.70 (dd, J = 11.0, 3.4 Hz, 1H, NCH), 3.50 (d, J = 11.7 Hz, 2H, ArCH2), 3.47 (s, 3H, NCH3), 3.04–3.02 (m, 1H, CH2), 2.83–2.82 (m, 1H, CH2), 2.38–2.31 (m, 1H, CH2), 1.99–1.83 (m, 3H, CH2), 1.79–1.74 (m, 2H, CH2). 13 13C NMR (75 MHz, Chloroform-d) δ 165.10, 159.98, 144.00, 142.81, 142.43, 141.40, 133.74, 132.95, 132.65, 131.40, 130.77, 129.13, 128.71, 127.35, 122.13, 121.56, 119.76, 113.78, 109.67, 103.67, 64.86, 61.84, 58.33, 55.46, 54.50, 32.81, 27.59, 23.49. HRMS-ESI (m / z): [M+H] + calcd for [C 32 H 33 N4O3] + 521.25472, found 521.25304, ppm error 2.92. m.p., 148.8~149.0 °C.

[0131] Example 58: The synthesis procedure of the target compound 1a-24 was referred to Example 35, using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-methyl-4-hydroxypiperidine as raw materials, and 27.5 mg of the product was obtained as a pale yellow solid with a yield of 41.55%. 11H NMR (400 MHz, Chloroform-d) δ 9.06 (s, 1H, ArH), 8.80 (s, 1H, NH), 8.25 (d, J = 7.8 Hz, 1H, ArH), 7.94 (d, J = 8.1 Hz, 2H, ArH), 7.61 (t, J = 7.4 Hz, 1H, ArH), 7.56 (d, J = 8.6 Hz, 2H, ArH), 7.47 (d, J = 7.6 Hz, 2H, ArH), 7.39 (d, J = 8.3 Hz, 1H, ArH), 7.30 (t, J = 7.5 Hz, 1H, ArH), 7.07 (d, J = 8.6 Hz, 2H, ArH), 3.91 (s, 3H, OCH3), 3.61 (s, 2H, ArCH2), 3.47 (s, 3H, NCH3), 2.58–2.44 (m, 4H, CH2), 1.73–1.68 (m, 4H, CH2), 1.24 (s, 3H, CH3). 13 13C NMR (75 MHz, Chloroform-d) δ 165.26, 159.99, 143.99, 143.10, 142.52, 141.35, 133.42, 132.93, 132.66, 131.47, 130.78, 129.27, 128.69, 127.17, 122.15, 121.57, 119.75, 109.65, 103.64, 77.25, 67.86, 62.72, 55.48, 49.88, 38.76, 32.79. HRMS-ESI (m / z): [M+H] + calcd for [C 33 H 35 N4O3] + 535.27037, found 535.26863, ppm error 2.81. m.p., 238.7~238.9 °C.

[0132] Example 59: The synthesis procedure of the target compound 1a-25 was referred to Example 35. Using 4-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 1-(4-nitrophenyl)piperazine as raw materials, 37.5 mg of the product was obtained as a pale yellow solid with a yield of 48.23%. 11H NMR (400 MHz, Chloroform-d) δ 9.09 (s, 1H, ArH), 8.94 (s, 1H, NH), 8.25 (d, J = 7.8 Hz, 1H, ArH), 8.12 (d, J = 9.4 Hz, 2H, ArH), 7.99 (d, J = 8.1 Hz, 2H, ArH), 7.61 (t, J = 7.7 Hz, 1H, ArH), 7.57 (d, J = 8.6 Hz, 2H, ArH), 7.50 (d, J = 8.1 Hz, 2H, ArH), 7.40 (d, J = 8.3 Hz, 1H, ArH), 7.31 (t, J = 7.5 Hz, 1H, ArH), 7.08 (d, J = 8.7 Hz, 2H, ArH), 6.82 (d, J = 9.4 Hz, 2H, ArH), 3.91 (s, 3H, OCH3), 3.65 (s, 2H, ArCH2), 3.47 (s, 3H, NCH3), 3.46–3.44 (m, 4H, CH2), 2.64–2.61 (m, 4H, CH2). 13 13C NMR (75 MHz, Chloroform-d) δ 165.15, 160.08, 154.82, 144.10, 142.34, 141.77, 141.21, 138.40, 133.80, 132.87, 132.82, 130.99, 130.80, 129.31, 128.89, 127.38, 125.95, 122.15, 121.48, 119.87, 113.82, 112.66, 109.75, 103.70, 62.34, 55.47, 52.54, 46.95, 32.84. HRMS-ESI (m / z): [M+H] + calcd for [C 37 H 35 N6O4] + 627.27143, found 627.27110, ppm error 0.41. m.p., 122.3~122.4 °C.

[0133] Example 60: The synthesis procedure of the target compound 1a-26 was referred to Example 35. Using 2-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxypiperidine as raw materials, 21.6 mg of the product was obtained as a white solid with a yield of 28.64%. 11H NMR (400 MHz, Chloroform-d) δ 13.02 (s, 1H, NH), 9.09 (s, 1H, ArH), 8.25 (d, J = 7.8 Hz, 1H, ArH), 8.13 (d, J = 7.5 Hz, 1H, ArH), 7.58 (t, J = 7.7 Hz, 1H, ArH), 7.52 (d, J = 8.6 Hz, 2H, ArH), 7.47 (t, J = 7.4 Hz, 1H, ArH), 7.42 (d, J = 6.3 Hz, 1H, ArH), 7.37 (d, J = 8.4 Hz, 1H, ArH), 7.29 (d, J = 7.4 Hz, 1H, ArH), 7.20 (d, J = 7.3 Hz, 1H, ArH), 7.02 (d, J = 8.6 Hz, 2H, ArH), 3.89 (s, 3H, OCH3), 3.65 (s, 2H, ArCH2), 3.59 (s, 1H, OH), 3.46 (s, 3H NCH3), 2.83 (s, 2H, CH2), 2.29–2.25 (m, 2H, CH2), 1.76–1.73 (m, 2H, CH2), 1.60–1.56 (m, 3H, CH2, CH). 13 13C NMR (75 MHz, Chloroform-d) δ 166.71, 159.65, 143.76, 143.74, 141.34, 137.75, 133.98, 132.78, 132.27, 132.21, 132.06, 131.60, 130.99, 130.21, 128.41, 128.35, 122.11, 121.80, 119.53, 113.18, 109.53, 104.37, 62.14, 55.41, 33.87, 32.78, 29.73, 26.93. HRMS-ESI (m / z): [M+H] + calcd for [C 32 H 33 N4O3] + 521.25472, found 521.25363, ppm error 1.80. m.p., 128.9~129.0 °C.

[0134] Example 61: The synthesis procedure of the target compound 1a-27 was referred to Example 35. Using 3-(chloromethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxypiperidine as raw materials, 26.3 mg of the product was obtained as a white solid with a yield of 39.25%. 11H NMR(500 MHz, DMSO-d6) δ 10.66 (s, 1H, NH), 8.81 (s, 1H, ArH), 8.28 (d, J = 7.8 Hz, 1H, ArH), 7.99 (s, 1H, ArH), 7.96 (d, J = 7.6 Hz, 1H, ArH), 7.63–7.59 (m, 4H, ArH), 7.50 (d, J = 7.4 Hz, 1H, ArH), 7.45 (t, J = 7.5 Hz, 1H, ArH), 7.30 (t, J = 6.6 Hz, 1H, ArH), 7.11 (d, J = 8.5 Hz, 2H, ArH), 4.58 (s, 1H, OH), 3.86 (s, 3H, OCH3), 3.52 (s, 2H, ArCH2), 3.46 (s, 3H, NCH3), 2.70–2.68 (m, 2H, CH2), 2.09–2.06 (m, 2H, CH2), 1.72–1.70 (m, 2H, CH2), 1.43–1.29 (m, 3H, CH, CH2). 13 13C NMR(126 MHz, DMSO-d6) δ 166.04, 159.90, 143.96, 143.04, 141.95, 139.33, 134.83, 132.69, 132.59, 131.82, 131.57, 131.54, 129.23, 128.86, 128.68, 126.94, 122.05, 121.12, 120.18, 113.85, 110.95, 105.15, 66.72, 62.31, 55.76, 51.34, 34.80, 33.14. HRMS-ESI (m / z): [M+H] + calcd for [C 32 H 33 N4O3] + 521.25472, found 521.25334, ppm error 2.21. m.p., 241.4~241.5 °C.

[0135] Example 62: The synthesis procedure of the target compound 1a-28 was referred to Example 35, using 4-(chloromethyl)-N-(1,9-dimethyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxypiperidine as raw materials, and 88.2 mg of the product was obtained as a white solid with a yield of 67.99%. 11H NMR(400MHz,DMSO-d6)δ10.55(s,1H,NH),8.68(s,1H,ArH),8.20(t,J=7.7Hz,1H,ArH),8.04(d,J=8.0Hz,2H,ArH),7.69(d,J=8.4Hz,1H,ArH),7.60(t,J=7.7Hz,1H,ArH),7.42(d,J=8.0Hz,2H,ArH),7.25(t,J=7.4Hz,1H,ArH),4.57(d,J=4.0Hz,1H,OH),4.15(s,3H,CH3),3.51(s,2H,ArCH2),3.46(s,1H,CH),3.02(s,3H,NCH3),2.69–2.66(m,2H,CH2),2.08–2.03(m,2H,CH2),1.73–1.70(m,2H,CH2),1.45–1.35(m,2H,CH2). 13 13C NMR(75MHz,DMSO-d6)δ165.48,143.14,143.06,142.77,140.43,133.53,133.35,130.23,128.93,128.74,128.24,121.88,120.90,119.66,110.63,103.93,62.18,51.45,34.92,32.46,22.99.HRMS-ESI(m / z):[M+H] + calcd for[C 26 H 29 N4O2] + 429.22850,found429.22702,ppm error3.10.m.p.,233.9~234.0℃.

[0136] Example62:The synthesis procedure of the target compound 1a-28 was referred to Example 35,using 4-(chloromethyl)-N-(9-isopropyl-1-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide and 4-hydroxypiperidine as raw materials to obtain 56.3mg of the product as a white solid in a yield of 50.97%. 11H NMR(300MHz,DMSO-d6)δ10.53(s,1H,NH),8.66(s,1H,ArH),8.21(d,J=7.7Hz,1H,ArH),8.05(d,J=7.8Hz,2H,ArH),7.85(d,J=8.5Hz,1H,ArH),7.54(t,J=7.8Hz,1H,ArH),7.43(d,J=6.5Hz,2H,ArH),7.24(t,J=7.5Hz,1H,ArH),5.48(p,J=7.0Hz,1H,CH),4.56(s,1H,OH),3.52–3.48(m,3H,ArCH2,CH),2.97(s,3H,OCH3),2.73–2.66(m,2H,CH2),2.09–2.04(m,2H,CH2),1.74–1.64(m,8H,CH3,CH2),1.48–1.5(m,2H,CH2). 13 13C NMR(75MHz,DMSO-d6)δ165.47,142.64,140.72,139.61,133.21,130.52,129.11,128.29,122.54,122.12,119.61,113.90,103.77,51.43,48.28,24.31,21.47.HRMS-ESI(m / z):[M+H] + calcd for[C 28 24H3N4O2] + 457.25980,found 457.25907,ppm error 0.93.m.p.,179.9~180.0℃.

[0137] III. Effect verification

[0138] Example 63: Study on the activity against protoscoleces (PSCs) of Echinococcus granulosus in vitro

[0139] 1. Experimental method: Under the conditions of 37 °C and 5% CO2, about 250 PSCs were inoculated into each well of a 96-well plate, with three replicates in each group. The target compound was dissolved in DMSO and then added to each well (containing 200 μL of medium) to make the final concentration of the compound 1, 5, 10 μM, and incubated for two days. PSCs were incubated in the medium containing 1% DMSO as the control group. Through a 1% eosin exclusion experiment, the effect of the target compound on the morphology and structural integrity of PSCs was observed under an optical microscope to evaluate its killing effect.

[0140] 2. Experimental results: To study whether the target compound can inhibit the growth of PSCs, we inoculated the protoscoleces of Echinococcus granulosus on 96-well plates at 37 °C and 5% CO2, and intervened with the target compound at different concentrations (1 μM, 5 μM, 10 μM), and incubated for 2 days for preliminary screening. The results are as Figure 1 shown in Figure Figure 1 and Table 1. Compounds 1a-1, 1a-2 and 1a-17 had strong insecticidal ability, and they caused 84.93 ± 2.22%, 98.58 ± 1.37%, 83.67 ± 11.39% death of Echinococcus granulosus at the dose of 10 μM, respectively. At the dose of 1 μM, the insecticidal effect of 1a-1 was the best, which was 34.28 ± 4.25%, equivalent to the lead compound 1a. The remaining compounds had no obvious insecticidal effect.

[0141] Table 1. In vitro insecticidal activities of the target compounds at different doses (1, 5 and 10 μM) a

[0142] Compound 1μM(%) 5μM(%) 10μM(%) Compound 1μM(%) 5μM(%) 10μM(%) Blank NA NA NA 1a-14 NA NA NA 1% DMSO NA NA NA 1a-15 NA NA NA 1a 45.34±7.34 53.07±7.51 86.40±2.13 1a-16 4.75±1.00 8.25±2.70 43.83±2.27 1a-1 34.28±4.25 50.80±5.42 84.93±2.22 1a-17 27.33±3.40 41.25±2.54 86.40±1.37 1a-2 15.11±2.55 57.11±4.99 83.67±11.39 1a-18 6.25±1.50 7.08±1.76 29.50±4.27 1a-3 10.00±3.50 17.17±3.06 28.67±4.31 1a-19 5.83±1.70 11.42±2.74 16.83±2.01 1a-4 NA NA 38.05±5.85 1a-20 NA NA NA 1a-5 NA NA NA 1a-21 2.00±0.66 3.75±1.15 11.17±1.62 1a-6 NA NA NA 1a-22 6.17±2.02 8.58±1.51 20.67±2.18 1a-7 10.96±2.26 35.75±5.75 71.11±6.78 1a-23 2.33±0.38 4.50±0.75 8.08±2.89 1a-8 NA NA NA 1a-24 6.83±2.25 11.50±1.64 26.08±2.74 1a-9 NA NA NA 1a-25 NA NA NA 1a-10 NA NA NA 1a-27 NA NA NA 1a-11 NA NA NA 1a-26 NA NA NA 1a-12 NA NA NA 1a-28 6.33±3.21 10.50±3.12 13.83±4.65 1a-13 NA NA NA 1a-29 7.67±1.15 13.17±5.11 17.67±4.16

[0143] a NA = no activity.

[0144] Example 64: In vitro stability study

[0145] 1. Experimental method: Weigh a certain mass of compound 1a, 1a-1 or 1a-2, dissolve it in 40 μL DMSO solution, and prepare a DMSO mother solution of a certain concentration. Add 3960 μL SD rat plasma to the DMSO mother solution, mix well, and observe whether the mixed solution is clear. If it is not clear, add Solutol (1% each time until it is clear). Place the mixed solution in a shaker and incubate at 37°C and 100-125 rpm / min. Sample 200 μL of the mixed solution at 0, 15, 30, 60, and 120 minutes in a 1.5 mL EP tube, add 200 μL methanol, mix well, centrifuge for 2 minutes, absorb the supernatant, filter, and determine the residual amount of the compound by high performance liquid chromatography (HPLC) (the compound peak area corresponding to 0 minutes is 100%). The residual amount of the compound is calculated as follows: residual amount = compound peak area at each time point / compound peak area at 0 minutes × 100%. The stability experiment of the compound in liver microsomes is similar to the operation method of the plasma stability experiment. A certain mass of compound 1a, 1a-1 or 1a-2 is weighed and dissolved in 10 μL DMSO solution to prepare a DMSO mother solution of a certain concentration. 910 μL SD rat plasma, 50 μL solution A, and 10 μL solution B are added to the DMSO mother solution to observe whether the mixed solution is clear (if the solution is not clear, shake the mixed solution on a shaker for about 5 minutes), then add 20 μL liver microsomes, mix well, incubate, sample at the corresponding time point, quench, centrifuge, take the supernatant, filter, and determine the remaining amount of the compound in the filtrate by HPLC.

[0146] 2. Experimental results: The target compound was incubated in rat plasma or buffer containing liver microsomes at 37°C for 120 minutes. Figure 2 As shown, the lead compound 1a was very easily metabolized in the presence of rat plasma or liver microsomes, with 29.10% and 7.73% remaining after 120 minutes of incubation, respectively. In contrast, compounds 1a-1 and 1a-2 were relatively stable, with 66.16% and 81.08% remaining after 120 minutes of incubation in rat plasma, and 86.15% and 91.78% remaining after 120 minutes of incubation in the presence of liver microsomes, respectively. The above results indicate that modification of the 9-position of β-carboline helps to improve the metabolic stability of the compound.

[0147] Example 65: Pharmacokinetic study in rats

[0148] 1. Experimental method: Weigh a certain mass of the target compound into an EP tube, add DMSO and solutol, and ultrasonicate and shake to mix evenly until the compound dissolves into a milky white homogeneous suspension. Then add Saline, ultrasonicate and shake to mix evenly and adjust the pH to 3. The compound becomes a yellowish-green clear state, and a test sample with a certain concentration is prepared, which is used as the intragastric administration preparation. SPF-grade male SD rats (n = 3) are given the derivatives (10 and 25 mg / kg) through intravenous or intragastric administration routes. Samples are taken at specific time points, and the concentration of the compound in plasma is determined by LC-MS / MS. Microsoft Excel 2010 spreadsheet software is used for data management, and WinNolin 8.2 software is used to calculate pharmacokinetic parameters.

[0149] Table 2. Main pharmacokinetic parameters of compounds 1a, 1a-1, and 1a-2 after intragastric administration in rats (n = 3) a

[0150] Parameters 1a 1a-1 1a-2 <![CDATA[T 1 / 2 (h)]]> ND 3.30±0.08 4.89±0.74 <![CDATA[T max (h)]]> ND 5.33±0.00 2.33±1.53 <![CDATA[C max (ng / mL)]]> ND 241±59.60 56.10±8.38 <![CDATA[AUC 0-t (h·ng / mL)]]> ND 2734±486 443±67.40 <![CDATA[AUC 0-∞ (h·ng / mL)]]> ND 2768±489 459±80.10 <![CDATA[MRT 0-t (h)]]> ND 7.52±0.04 6.05±1.08 <![CDATA[MRT 0-∞ (h)]]> ND 7.78±0.07 6.91±1.64 F(％) ND 27.80％ 6.01％

[0151] ND=Not Detected

[0152] 2. Experimental results: Compounds 1a-1 and 1a-2 were administered to healthy male SD rats by intragastric administration (dose 25 mg / kg) and intravenous injection (dose 10 mg / kg), and samples were taken at different time points. The concentrations of compounds 1a-1 and 1a-2 in rat plasma samples were determined by LC-MS / MS, and WinNolin software was used to calculate pharmacokinetic parameters. The results showed ( Figure 3 , Table 2) that after intragastric administration of 1a-1, C max was 241 ng / mL, T max was 5.33 h, T 1 / 2 was 3.30 h, AUC 0-t was 2734 hr·ng / mL, and the oral bioavailability was 27.80%. After intragastric administration of 1a-2, C max was 56.1 ng / mL, T max was 2.33 h, T 1 / 2 was 4.89 h, AUC 0-t was 443 hr·ng / mL, and the oral bioavailability was 6.01%. However, when 1a was orally administered at doses of 50 mg / kg and 100 mg / kg, the blood drug concentration of 1a could not be measured, and the pharmacokinetic parameters after oral administration of 1a could not be calculated. It can be seen that by modifying the lead compound 1a, its analog 1a-1 has better pharmacokinetic properties and significantly improves the oral bioavailability.

[0153] Example 66: In Vivo Pharmacodynamic Study in Mice

[0154] 1. Experimental method: In the in vivo experiment, pathogen-free female KM mice at 6 - 8 weeks old were purchased from the Experimental Animal Center of Xinjiang Medical University. The use of experimental animals was approved by the Ethics Committee of the First Affiliated Hospital of Xinjiang Medical University (Approval number: ACUC - 20170420 - 04).

[0155] PSCs were pre - cultured in vitro to produce small cysts (micro - cysts, diameter 200 - 300 μm), and were inoculated into 6 - 8 - week - old KM mice by intraperitoneal injection. For the preparation of micro - cysts, PSCs digested with 1% pepsin (viability > 98%) were cultured in culture flasks containing RPMI 1640 medium (medium containing 10% (v / v) fetal bovine serum, 100 IU / mL penicillin and 100 μg / mL streptomycin). The culture flasks were placed in an incubator at 37 °C and 5% CO2, and the medium was changed every 4 - 5 days. PSCs were pre - cultured in vitro for 3 - 4 months. Each mouse was intraperitoneally inoculated with 25 micro - cysts (suspended in 0.4 mL of physiological saline). Approximately 180 days after infection, the model was detected by B - ultrasound. The model mice were randomly divided into 10 groups (n = 6 in each group): (1) The model group was given physiological saline (0.1 mL / 10 g / day); (2) The ABZ (high, medium, low dose) groups were given ABZ suspension at 50, 25, 12.5 mg / kg / day respectively; (3) The HM (high, medium, low dose) groups were given HM solution at 50, 25 and 12.5 mg / kg / day respectively; (3) The 1a - 1 group (high, medium, low dose) groups were provided with 1a - 1 solution at doses of 50, 25, 12.5 mg / kg / day respectively. The experimental mice were continuously administered by gavage or intraperitoneal injection for 28 days. During the treatment period, the body weight changes and survival of the mice were monitored daily. After the administration ended, the mice were sacrificed, and the peritoneal cysts of the mice were separated and weighed. The formula for calculating the wet weight of the cysts is as follows: (Average cyst weight of the model group - Average cyst weight of the treatment group) / Average cyst weight of the model group × 100%.

[0156] 2. Experimental results: To study the therapeutic effect of compound 1a - 1 on hydatid cysts in vivo, we infected KM mice with a secondary hydatid mouse model by intraperitoneal injection for a 6 - month infection period. All model mice were randomly divided into 10 groups on average, including a model group and dosing groups. The dosing groups were divided into three doses: high (50 mg / kg / day), medium (25 mg / kg / day), and low (12.5 mg / kg / day).

[0157] Table 3 - 4. Therapeutic effect of compound 1a - 1 on mice infected with Echinococcus granulosus (28 days) (n = 6)

[0158]

[0159]

[0160] The results are as Figure 4 shown in Table 3. After intragastric administration of compound 1a-1 at high, medium, and low doses for 28 days, the cyst inhibition rates were 77.67%, 70.57%, and 64.63% respectively, which were comparable to the in vivo therapeutic effects of the lead compound 1a (intraperitoneal injection at high, medium, and low doses, with cyst inhibition rates of 76.87%, 72.05%, and 67.17% respectively), and were superior to ABZ and HM at the same doses, showing significant differences. In addition, at the low dose, the therapeutic effect of compound 1a-1 was better than that of the high-dose groups of ABZ and HM.

[0161] Example 67: Pharmacokinetics study in sheep

[0162] 1. Experimental method: Weigh a certain mass of compound 1a-1 into an EP tube, add DMSO and solutol, and ultrasonicate and shake to mix until the compound dissolves into a milky white homogeneous suspension. Then add Saline and ultrasonicate and shake to adjust the pH to 3, and the compound becomes a yellowish-green clear state, and a test sample with a certain concentration (2.5 mg / kg by intravenous injection, 25 mg / kg by intragastric administration) is prepared. This solution is used as the intragastric and intravenous administration preparations. Healthy sheep (n = 3) were administered compound 1a-1 through intravenous (2.5 mg / kg), single intragastric (25 mg / kg), and continuous intragastric administration for three days (25 mg / kg, once a day). Samples were taken at specific time points (intravenous: 0.0833, 0.167, 0.333, 0.5, 1, 2, 4, 6, 8, 12, 24 h; single intragastric: 0.0833, 0.250, 0.50, 1.0, 2, 4, 6, 8, 12, 24, 36, 48 h; continuous intragastric administration for three days: 0.250, 0.50, 1.0, 2, 4, 6, 8, 12, 24, 36, 48, 60, 72, 84, 96 h), and the concentration of the compound in plasma was measured by LC-MS / MS. Microsoft Excel 2010 spreadsheet software was used for data management, and WinNolin 8.2 software was used to calculate the pharmacokinetic parameters.

[0163] 2. Experimental results: The results showed ( Figure 5 , Table 4) that after intravenous injection of 1a-1, C max was 251 ng / mL, T max was 0.0833 h, T 1 / 2 was 12.2 h, and Cl was 42.6 mL / hr / kg. After single intragastric administration of 1a-1, C max was 107 ng / mL, T max was 40.0 h, AUC0-t was 3084 hr·ng / mL, and the oral bioavailability was 36.90%. After intragastric administration of 1a-1 for three consecutive days, C max was 324 ng / mL, T max was 28 h, T 1 / 2 was 46.5 h, AUC 0-t was 19631 hr·ng / mL, showing a cumulative effect. It can be seen that compound 1a-1 has good pharmacokinetic properties in sheep and a high oral bioavailability.

[0164] Table 4. Main pharmacokinetic parameters of compound 1a-1 in healthy sheep (n = 3)

[0165]

[0166]

[0167] Example 68: Pharmacodynamic study in sheep

[0168] 1. Experimental method: In the pharmacodynamic experiment in sheep, go to the Bayinbulak natural pasture in Xinjiang Uygur Autonomous Region, and screen sheep infected with Echinococcus granulosus by B-ultrasound screening and Raman spectroscopy-assisted methods. Then isolate and transfer the infected sheep to a specific breeding area for breeding. Measure the cyst size in the sheep by CT scan, and randomly divide the infected sheep into four groups: model group, albendazole (ABZ) group (25 mg / kg), 1a-1 intragastric administration group (25 mg / kg), 1a-1 intravenous administration group (2.5 mg / kg). The infected sheep were intragastrically administered or intravenously injected for 14 consecutive days. During the treatment period, monitor the weight change and survival of the sheep, and monitor the change of vesicle size before and after drug intervention by B-ultrasound. After the treatment, sacrifice and dissect the sheep, collect tissue samples for detection and calculate the cyst inhibition rate to evaluate the efficacy.

[0169] 2. Experimental results: The results are as Figure 6 shown. Monitor the weight and survival of the sheep every week. During the entire experimental period of the model group, 1a-1 oral administration group and 1a-1 intravenous administration group, the mental state, feeding, drinking, breathing and activities of the infected sheep were normal, and the weight showed an upward trend. However, after 7 days of administration in the ABZ oral group, the sheep showed symptoms such as anorexia and diarrhea, and there was a significant difference in the weight of the sheep on the 14th day compared with that on the 0th day. By comparing between groups, it was found that there was no significant difference in the weight of the sheep among groups on the 0th day. After 7 days of administration, there was a significant difference in the weight between the ABZ oral group and the model group. After 14 days of administration, there were significant differences between the ABZ oral group and the other groups.

[0170] The volume change of vesicles in infected sheep was monitored by B-ultrasound. In the model group, the growth rate of vesicle volume in sheep showed an upward trend. Compared with the 7th day, the growth rate of vesicle volume was significantly increased on the 14th day. The growth rate of vesicles in the drug administration group as a whole showed a downward trend. There was no significant change in the growth rate of vesicle volume after 7 days and 14 days of intervention in the ABZ oral administration group. The growth rate of vesicle volume reached the minimum after 14 days of intervention in the 1a-1 oral administration group and the intravenous administration group, and there was a significant difference compared with that after 7 days of intervention. Comparison among groups found that after 7 days of treatment, the growth rate of vesicle volume in sheep in each drug administration group was significantly lower than that in the model group; after 14 days of treatment, the growth rate of vesicle volume in sheep in the 1a-1 oral administration group and the intravenous administration group reached the minimum, and there was a significant difference in the growth rate of vesicle volume between the 1a-1 intravenous administration group and the ABZ oral administration group. After 14 days of treatment, we dissected the sheep for overall observation. The vesicles in the model group were full and clear. The vesicles in the ABZ oral administration group were mostly semi-transparent. The vesicles in the 1a-1 oral administration group were milky yellow or calcified nodules with relatively hard texture. The vesicles in the 1a-1 intravenous administration group were significantly reduced, the vesicle boundary was blurred, and there were multiple calcification points, showing an obvious calcification trend. Compared with the model group, the vesicle volume in sheep in the ABZ oral administration group and its 1a-1 oral and intravenous administration groups was significantly reduced; among them, there was a significant difference in the vesicle volume between the 1a-1 intravenous administration group and the ABZ oral administration group.

Claims

1. A compound, characterized in that A compound represented by formula Ⅰ, Ⅱ, Ⅲ, Ⅳ, or a pharmaceutically acceptable salt thereof: Among them, R 1 Selected from -H, -CH3, -CH2CH3, -CH2CH2CH3; R 2 Selected from n is 1, 2 or 3; R 3 Selected from In formula III The substitution position on the benzene ring is ortho or meta; R 4 Selected from -CH3, -CH(CH3)2.

2. The compound according to claim 1, characterized in that The compound represented by formula I is selected from: 1a-1: 4-((4-hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-2: N-(9-ethyl-1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-((4-hydroxypiperidin-1-yl)methyl)benzamide 1a-3: 4-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-propyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-4: 4-(4-Hydroxypiperidin-1-yl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-5: N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-(piperidin-1-yl)benzamide 1a-6: N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-morpholinylbenzamide 1a-7: N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide 1a-8: 4-(Diethylamino)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-9: N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-(pyrrolidin-1-yl)benzamide 1a-10: 4-(dimethylamino)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-11: N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-(piperidin-1-yl)benzamide 1a-12: N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-morpholinylbenzamide 1a-13: 4-(Diethylamino)-N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-14: 4-(Dimethylamino)-N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-15: N-(1-(4-methoxyphenyl)-9H-pyrido[3,4-b]indol-3-yl)-4-(4-methylpiperazin-1-yl)benzamide 3. The compound according to claim 1, characterized in that The compound shown in formula II is selected from: 1a-16: 4-((4-(Hydroxymethyl)piperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-17: 4-((4-(2-hydroxyethyl)piperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-18: 4-((4-(3-hydroxypropyl)piperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-19: 4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-20: 4-((4-(3-hydroxypropyl)piperazin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-21: 4-(2-(4-hydroxypiperidin-1-yl)ethyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-22: 4-(3-(4-hydroxypiperidin-1-yl)propyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-23: (S)-4-((2-(Hydroxymethyl)pyrrolidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-24: 4-((4-hydroxy-4-methylpiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-25: N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)-4-((4-(4-nitrophenyl)piperazin-1-yl)methyl)benzamide 4. The compound according to claim 1, characterized in that The compound shown in formula III is selected from: 1a-26: 2-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 1a-27: 3-((4-Hydroxypiperidin-1-yl)methyl)-N-(1-(4-methoxyphenyl)-9-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 5. The compound according to claim 1, characterized in that The compound shown in formula IV is selected from: 1a-28: N-(1,9-dimethyl-9H-pyrido[3,4-b]indol-3-yl)-4-((4-hydroxypiperidin-1-yl)methyl)benzamide 1a-29: 4-((4-Hydroxypiperidin-1-yl)methyl)-N-(9-isopropyl-1-methyl-9H-pyrido[3,4-b]indol-3-yl)benzamide 6. The method for preparing the compound according to claim 1, characterized in that: The preparation method of compound I comprises: The raw material L-tryptophan and 4-methoxybenzaldehyde undergo Pictet-Spengler reaction under acidic conditions to obtain intermediate 1-1; the carboxyl group of intermediate 1-1 is converted into methyl ester to obtain intermediate 1-2, which is subsequently oxidized to obtain intermediate 1-3; intermediate 1-3 undergoes nucleophilic substitution reaction with compound 1-4 to obtain intermediate 1-5; intermediate 1-5 reacts with hydrazine hydrate to obtain intermediate 1-6, the hydrazine group of intermediate 1-6 is converted into intermediate 1-7 in the presence of sodium nitrite, intermediate 1-7 undergoes Curtis rearrangement reaction to obtain intermediate 1-8, intermediate 1-8 undergoes amide condensation reaction with compound 1-9 to obtain target compound I; or, intermediate 1-8 reacts with 4-chloromethylbenzoyl chloride to obtain intermediate 1-10, and intermediate 1-10 reacts with 4-hydroxypiperidine to obtain target compound I.

7. The method for preparing the compound according to claim 1, characterized in that: The preparation method of compound II comprises: The raw material L-tryptophan and 4-methoxybenzaldehyde undergo Pictet-Spengler reaction under acidic conditions to obtain intermediate 2-1, the carboxyl group of intermediate 2-1 is converted into methyl ester to obtain intermediate 2-2, which is subsequently oxidized to obtain intermediate 2-3; intermediate 2-3 undergoes nucleophilic substitution reaction with methyl iodide to obtain intermediate 2-4, intermediate 2-4 reacts with hydrazine hydrate to obtain intermediate 2-5, the hydrazine group of intermediate 2-5 is converted into intermediate 2-6 in the presence of sodium nitrite, intermediate 2-6 undergoes Curtis rearrangement reaction to obtain intermediate 2-7, intermediate 2-7 and compound 2-8 undergo amide condensation reaction to obtain intermediate 2-9; intermediate 2-9 reacts with 4-hydroxypiperidine to obtain the target compound II; or, intermediate 2-7 reacts with 4-chloromethylbenzoyl chloride to obtain intermediate 2-10; intermediate 2-10 reacts with compound 2-11 to obtain the target compound II.

8. The method for preparing the compound according to claim 1, characterized in that: The preparation method of compound III comprises: The raw material L-tryptophan and 4-methoxybenzaldehyde undergo Pictet-Spengler reaction under acidic conditions to obtain intermediate 3-1, the carboxyl group of intermediate 3-1 is converted into methyl ester to obtain intermediate 3-2, which is subsequently oxidized to obtain intermediate 3-3; intermediate 3-3 undergoes nucleophilic substitution reaction with methyl iodide to obtain intermediate 3-4, intermediate 3-4 reacts with hydrazine hydrate to obtain intermediate 3-5, the hydrazine group of intermediate 3-5 is converted into intermediate 3-6 in the presence of sodium nitrite, intermediate 3-6 undergoes Curtis rearrangement reaction to obtain intermediate 3-7, intermediate 3-7 undergoes amide condensation reaction with compound 3-8 to obtain intermediate 3-9, wherein the substitution position of -CH2Cl on the benzene ring in compound 3-8 is ortho or meta; intermediate 3-9 reacts with 4-hydroxypiperidine to obtain the target compound III.

9. The method for preparing the compound according to claim 1, characterized in that: The preparation method of compound IV comprises: The raw material L-tryptophan reacts with acetaldehyde under acidic conditions to undergo Pictet-Spengler reaction to obtain intermediate 4-1, the carboxyl group of intermediate 4-1 is converted into methyl ester to obtain intermediate 4-2, which is subsequently oxidized to obtain intermediate 4-3; intermediate 4-3 reacts with compound 4-4 to obtain intermediate 4-5 by nucleophilic substitution reaction, intermediate 4-5 reacts with hydrazine hydrate to obtain intermediate 4-6, the hydrazine group of intermediate 4-6 is converted into intermediate 4-7 in the presence of sodium nitrite, intermediate 4-7 undergoes Curtis rearrangement reaction to obtain intermediate 4-8, intermediate 4-8 undergoes amide condensation reaction with 4-chloromethylbenzoyl chloride to obtain intermediate 4-9; intermediate 4-9 reacts with 4-hydroxypiperidine to obtain the target compound IV.

10. Use of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating echinococcosis.

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