CN116675689A

CN116675689A - Beta-carboline derivative and preparation method and application thereof

Info

Publication number: CN116675689A
Application number: CN202310499454.7A
Authority: CN
Inventors: 徐学涛; 肖狄; 梁冰雯
Original assignee: Wuyi University
Current assignee: Wuyi University
Priority date: 2023-05-05
Filing date: 2023-05-05
Publication date: 2023-09-01

Abstract

The invention discloses a beta-carboline derivative or pharmaceutically acceptable salt thereof, and a preparation method and application thereof, wherein the beta-carboline derivative has a structure shown in a formula (I):wherein R is selected from the group consisting of substituted and unsubstituted C _6～12 Aryl, substituted or unsubstituted C _5～12 Heteroaryl of (a). The invention designs and synthesizes a series of beta-carboline compounds by taking the derivative structure of the beta-carboline as a parent nucleus, and the compounds all show strong alpha-glucosidase inhibition effect and IC ₅₀ Low in value, and can be used as an alpha-glucosidase inhibitor for treating or preventing diabetes.

Description

Beta-carboline derivative and preparation method and application thereof

Technical Field

The invention relates to the technical field of organic synthesis, in particular to a beta-carboline derivative, a preparation method and application thereof.

Background

The risk of diabetes is continuously rising due to the change of survival conditions, the change of dietary structure, and hyperglycemia is a metabolic disease due to the decrease or insufficiency of insulin secretion, mainly characterized by hyperglycemia. If the blood glucose level is not controlled timely, a series of dysfunction and damage can be induced, and the health and safety of patients are seriously threatened. Researches show that the diabetes has the characteristics of long course of disease, difficult cure and high disability rate, and most patients need long-term drug treatment. Diabetes is also classified into type I diabetes and type II diabetes. Of these, class i hyperglycemia, also known as insulin-dependent hyperglycemia (IDDM), is about ten percent of all hyperglycemia, mainly caused by severe impairment of insulin's islet β -cell function in humans, and thus by reduced endogenous insulin resistance secretion, and is inherited to some extent.

Type II diabetes, also known as non-insulin-dependent diabetes mellitus (NIDDM), is mostly caused by the action of genes and environmental factors, such as mutation of genes, overeating, decrease of exercise amount, etc. The pathogenesis is mainly caused by impaired insulin action, impaired insulin secretion or increased hepatic glucose production. Type ii diabetes is caused by insulin resistance, with adequate levels of insulin. Gestational diabetes usually occurs in pregnant women and disappears after birth. From the IDF diabetes atlas (7 th edition), it is estimated that about 4.15 million people have diabetes and that the proportion of 20-79 year old adults is 8.8%. In addition, more and more people are diagnosed with type ii diabetes due to unhealthy diet, limited exercise, and obesity. It is estimated that if this trend is continued until two to four years, one of every ten teenagers suffers from diabetes. Of the diabetics, it is estimated that about 87-91% of patients have type II diabetes and 7-12% have type I diabetes.

At present, the clinical medicines for treating diabetes are various in variety, and different medicines have different advantages and disadvantages in treatment effect. The clinic diabetes treatment mainly comprises insulin resistance secretagogues, insulin resistance sensitizers, biguanides, insulin resistance, insulin analogues and the like, can generate a certain blood glucose reducing effect, and is beneficial to effectively controlling the blood glucose level. Especially, with the continuous and intensive research on pathogenesis of the diabetes mellitus, new drugs are continuously emerging, and the new hypoglycemic drugs are achieved in the aspect of treating diabetes mellitus. It should be noted that when clinical medicines are used for treating diabetes, the conditions, medical history, medicine characteristics and specific conditions of patients are combined, and the hypoglycemic medicines are scientifically and reasonably selected, so that good hypoglycemic effect is realized, and adverse reactions are reduced. Although clinical diabetes medicine has various treatment types, the curative effect, action mechanism, adverse reaction and other aspects of the medicine need to be continuously researched, so that more choices are further provided for individual treatment of diabetes patients.

A glucosidase inhibitor is an oral hypoglycemic drug that can reach diagnostic blood glucose levels by reducing the rate of carbohydrate digestion, as well as controlling postprandial hyperglycemia levels. The action mechanism is as follows: alpha-glucosidase inhibitors are able to slowly release large amounts of D-glucose from oligosaccharides and disaccharides by affecting enzymatic function in the brush border of the small intestine, thereby causing a decrease in postprandial plasma glucose concentration and slowing down glucose digestion and absorption rate. Alpha-glucosidase is a family of enzymes located on brush border of small intestine cells, and is a carbohydrate inhibitor that specifically breaks down 1, 4-alpha-glucosidic bonds and emits a large amount of alpha-D-glucose. Alpha-glucosidase plays a key role in the whole body circulation of mammals, and it is based on this that inhibitors of this enzyme play a key role in diagnosing blood glucose levels.

Therefore, there is a need to develop new compounds having an effect of inhibiting α -glucosidase.

Disclosure of Invention

The present invention aims to solve at least one of the technical problems existing in the prior art. Therefore, the invention provides a beta-carboline derivative which can effectively inhibit alpha-glucosidase, thereby being used for treating and/or preventing diabetes.

The second aspect of the invention also provides a preparation method of the beta-carboline derivative.

The third aspect of the present invention also provides a pharmaceutical composition.

The fourth aspect of the invention also provides an alpha-glucosidase inhibitor.

The fifth aspect of the invention also provides an application of the beta-carboline derivative.

According to the embodiment of the first aspect of the invention, the beta-carboline derivative or the pharmaceutically acceptable salt thereof has a structure shown in a formula (I):

wherein R is selected from the group consisting of substituted and unsubstituted C _6～12 Aryl, substituted or unsubstituted C _5～12 Heteroaryl of (a).

The beta-carboline derivative or the pharmaceutically acceptable salt thereof provided by the embodiment of the invention has at least the following beneficial effects:

the invention designs and synthesizes a series of beta-carboline compounds by taking the derivative structure of the beta-carboline as a parent nucleus, and the compounds all show strong alpha-glucosidase inhibition effect and IC ₅₀ Has a low value of 239-985 times that of acarbose, and can be used as an alpha-glucosidase inhibitor for treating or preventing diabetes.

According to some embodiments of the invention, the R is selected from substituted or unsubstituted phenyl, the substitution is mono-or poly-substituted, the substituted group is selected from halogen, C _1～6 Alkyl, C of (2) _1～6 Alkoxy, C _1～6 Haloalkyl, hydroxy, nitro, cyano.

According to some embodiments of the invention, R is selected from one of the following groups:

according to a second aspect of the present invention, there is provided a method for preparing a β -carboline derivative or a pharmaceutically acceptable salt thereof, comprising the steps of:

carrying out condensation reaction on the compound 1 and an aldehyde compound containing an R group in a solvent to obtain a compound shown in a formula (I);

wherein, the structure of compound 1 is as follows:

according to some embodiments of the invention, the solvent is selected from at least one of ethanol, tetrahydrofuran, pyridine, methanol, or N, N-dimethylformamide.

According to some embodiments of the invention, the temperature of the reaction is 15-30 ℃.

According to some embodiments of the invention, the reaction further comprises a purification step after completion.

According to some embodiments of the invention, the purifying step comprises solid-liquid separation, recrystallization.

The third aspect of the invention provides a pharmaceutical composition comprising a β -carboline derivative according to any one of the preceding claims or a pharmaceutically acceptable salt thereof; pharmaceutically acceptable auxiliary materials.

According to a fourth aspect of the present invention there is provided an alpha-glucosidase inhibitor comprising a beta-carboline derivative as described in any preceding claim, or a pharmaceutically acceptable salt thereof.

The fifth aspect of the present invention provides the use of a β -carboline derivative according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of the preceding claims, in the manufacture of a medicament for the treatment and/or prophylaxis of diabetes.

Definitions and general terms

"substituted or unsubstituted C _6～12 Aryl "of (a) represents an all-carbon monocyclic or fused polycyclic group having a fully conjugated pi-electron system; and the total number of carbon atoms is 6 to 12. And optionally at least one H in the aryl group is substituted with a corresponding group as defined herein.

"substituted or unsubstituted C _5～12 The heteroaryl group "of (a) is represented as a monocyclic or fused ring group of ring atoms containing one, two, three or four ring heteroatoms selected from N, O or S, the remaining ring atoms being C, further having a fully conjugated pi-electron system and the total number of carbon atoms being 512. And optionally at least one H in the heteroaryl group is substituted with a corresponding group as defined herein.

“C _1～6 Alkyl "of (C) represents an alkyl group having 1 to 6 total carbon atoms including C _1-6 Straight chain alkyl, C _1-6 Branched alkyl and C of (2) _3-6 For example, a straight-chain alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, a branched-chain alkyl group having 1, 2, 3, 4, 5 or 6 carbon atoms, or a cycloalkyl group having 3, 4, 5 or 6 carbon atoms, and for example, a methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, tert-butyl group, n-pentyl group, isopentyl group, n-hexyl group, cyclopropyl group, methylcyclopropyl group, ethylcyclopropyl group, cyclopentyl group, methylcyclopentyl group, cyclohexyl group, or the like may be mentioned.

“C _1～6 Alkoxy "of (a) represents an alkoxy group having a total of 1 to 6 carbon atoms, including a linear alkoxy group having 1 to 6 carbon atoms, a branched alkoxy group having 1 to 6 carbon atoms and a cyclic alkoxy group having 2 to 6 carbon atoms, and may be, for example, a linear alkoxy group having 1, 2, 3, 4, 5 or 6 carbon atoms, a branched alkoxy group having 1, 2, 3, 4, 5 or 6 carbon atoms or a cyclic alkoxy group having 2, 3, 4, 5 or 6 carbon atoms, and may be, for example, methoxy, ethoxy, n-propoxy, isopropoxy and the like.

“C _1～6 "haloalkyl" represents an alkyl radical having a total number of carbon atoms of 1 to 6, preferably an alkyl radical as defined above, which is substituted by one or more identical or different halogen atoms, for example-CH ₂ Cl、-CF ₃ 、-CCl ₃ 、-CH ₂ CF ₃ 、-CH ₂ CCl ₃ Etc.

The structural appearance of the inventionRepresenting the attachment site of the group.

The term "pharmaceutically acceptable" as used herein refers to substances that are acceptable from a toxicological standpoint for pharmaceutical use and do not adversely interact with the active ingredient.

Pharmaceutically acceptable excipients for use in the present invention include any solvent, solid excipient, diluent, binder, disintegrant, or other liquid excipient, dispersing agent, flavoring or suspending agent, surfactant, isotonic agent, thickening agent, emulsifying agent, preservative, solid binder, glidant or lubricant, and the like, as appropriate for the particular target dosage form. As described in the following documents: in Remington, the Science and Practice of Pharmacy,21st edition,2005,ed.D.B.Troy,Lippincott Williams&Wilkins,Philadelphia,and Encyclopedia of Pharmaceutical Technology,eds.J.Swarbrick and J.C.Boylan,1988-1999,Marcel Dekker,New York, in combination with the teachings of the literature herein, shows that different excipients can be used In the preparation of pharmaceutically acceptable compositions and their well-known methods of preparation. In addition to the extent to which any conventional adjuvant is incompatible with the compounds of the present invention, such as any adverse biological effects produced or interactions with any other component of the pharmaceutically acceptable composition in a deleterious manner, their use is also contemplated by the present invention.

Materials that are pharmaceutically acceptable excipients include, but are not limited to, ion exchangers; aluminum; aluminum stearate; lecithin; serum proteins, such as human serum proteins; buffer substances such as phosphates; glycine; sorbic acid; potassium sorbate; a partial glyceride mixture of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silicon; magnesium trisilicate; polyvinylpyrrolidone; polyacrylate; a wax; polyethylene-polyoxypropylene-block polymers; lanolin; sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; a gum powder; malt; gelatin; talc powder; adjuvants such as cocoa butter and suppository waxes; oils such as peanut oil, cotton seed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycol compounds such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salt; ringer's solution; ethanol; phosphate buffer solution; and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate; a colorant; a release agent; coating the clothing material; a sweetener; a flavoring agent; a perfume; preservatives and antioxidants.

The pharmaceutical compositions of the compounds of the present invention may be administered in unit dosage form. The administration dosage form may be liquid dosage form or solid dosage form. The liquid dosage form can be true solution, colloid, microparticle, or suspension. Other dosage forms such as tablet, capsule, dripping pill, aerosol, pill, powder, solution, suspension, emulsion, granule, suppository, lyophilized powder for injection, etc.

Oral tablets and capsules may contain excipients such as binding agents, for example syrup, acacia, sorbitol, tragacanth or polyvinylpyrrolidone; fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, glycine; lubricants, such as magnesium stearate, talc, polyethylene glycol, silica; disintegrants, such as potato starch; or acceptable wetting agents such as sodium lauryl sulfate. The tablets may be coated by methods known in the pharmaceutical arts.

The oral liquid may be formulated as a suspension, solution, emulsion, syrup or elixir in a hydrated oil, or as a dry product, supplemented with water or other suitable medium prior to use. Such liquid preparations may contain conventional additives such as suspending agents, sorbitol, cellulose methyl ether, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel, hydrogenated edible fats and oils, emulsifying agents such as lecithin, sorbitan monooleate, acacia; or a non-aqueous carrier (possibly containing edible oils), such as almond oil, fats and oils such as glycerin, ethylene glycol, or ethyl alcohol; preservatives, such as methyl or propyl parahydroxybenzoate, sorbic acid. Flavoring or coloring agents may be added as desired.

Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.

Drawings

The foregoing and/or additional aspects and advantages of the invention will become apparent and may be better understood from the following description of embodiments taken in conjunction with the accompanying drawings in which:

FIG. 1 is a graph showing the half inhibition concentration of a beta-carboline derivative as an alpha-glucosidase inhibitor to alpha-glucosidase in vitro;

FIG. 2 is an in vitro enzymatic kinetic profile of β -carboline derivatives (X1-26) as α -glucosidase inhibitors of example 39 of the invention for α -glucosidase;

FIG. 3 is a graph showing the in vitro substrate kinetics of β -carboline derivatives X1-26 as α -glucosidase inhibitors against α -glucosidase in example 40 of the invention.

Detailed Description

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

The reagents, methods and apparatus employed in the present invention, unless otherwise specified, are all conventional in the art.

Examples 1 to 37

Examples 1 to 37 provide a series of beta-carboline derivatives, the general preparation method of which is as follows:

a series of aldehydes containing R groups (R-CHO) were added to 10mL of an absolute ethanol solution of the β -carboline hydrazone derivative (1 mmol) at normal temperature, and the mixture was stirred at room temperature and detected by TLC until the reaction was completed. Quenching by adding water, and filtering to obtain a crude product. The crude product is recrystallized by ethanol to obtain the compounds X1-1 to X1-37 with the yield of 65.3 percent to 87.8 percent.

The product obtainedThe method comprises the following steps:

wherein R of X1-1 to X1-37 are shown in the following Table 1:

TABLE 1 structural formula of R group

The structure of X1-1 to X1-37 was characterized by NMR, MS and melting point, and the properties, yields, nuclear magnetism and mass spectrum results of each compound were characterized as follows:

(X1-1,C ₂₅ H ₁₈ N ₄ O).Yellow sold；Yield 71％；m.p.347-357℃； ¹ H NMR(500MHz,DMSO-d6)δ11.95(s,1H),11.82(s,1H),8.97(s,1H),8.68(d,J＝2.3Hz,1H),8.48(d,J＝7.9

Hz,1H),8.25–8.20(m,2H),7.80–7.75(m,2H),7.74–7.65(m,4H),7.61(q,J＝7.6Hz,2H),

7.52–7.42(m,4H),7.34(t,J＝7.5Hz,1H)； ¹³ C NMR(126MHz,DMSO)δ161.27,148.33,141.61,140.99,139.02,137.38,134.57,134.55,130.02,129.92,129.06,128.99,128.89,128.86,

128.78,127.13,122.22,121.19,120.38,114.09,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcdfor C ₂₅ H ₁₈ N ₄ O:391.1552；found:391.1553.

(X1-2,C ₂₆ H ₂₀ N ₄ O).White sold；Yield 64％；m.p.349.5-366.1℃； ¹ H NMR(500MHz,DMSO-d6)δ11.93(s,1H),11.86(s,1H),8.96(d,J＝10.1Hz,2H),8.48(d,J＝7.8Hz,1H),

8.23–8.18(m,2H),7.93(dd,J＝7.5,1.7Hz,1H),7.70(q,J＝7.8Hz,3H),7.65–7.57(m,

2H),7.37–7.24(m,4H),2.49(s,3H)； ¹³ C NMR(126MHz,DMSO)δ161.34,146.84,141.61,141.07,139.17,137.40,137.03,134.57,132.64,130.85,129.87,129.67,129.03,129.00,128.89,

128.76,126.14,125.90,122.21,121.19,120.36,114.13,112.75,19.18；HRMS(ESI-MS)m/z:

[M+H] ⁺ calcd for C ₂₆ H ₂₀ N ₄ O:405.1708；found:405.1710.

(X1-3,C ₂₆ H ₂₀ N ₄ O).Yellow sold；Yield 65％；m.p.334.7-346℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(s,1H),11.75(s,1H),8.96(s,1H),8.63(s,1H),8.48(d,J＝7.9Hz,1H),

8.22(d,J＝7.3Hz,2H),7.79–7.65(m,6H),7.61(q,J＝7.6Hz,2H),7.34(t,J＝7.5Hz,1H),

7.29(d,J＝7.9Hz,2H),2.36(s,3H)； ¹³ C NMR(126MHz,DMSO)δ161.16,148.36,141.60,140.95,139.80,139.07,137.38,134.51,131.84,129.92,129.48,129.04,128.97,128.84,128.75,

127.11,122.21,121.19,120.36,114.02,112.74,21.07；HRMS(ESI-MS)m/z:[M+H] ⁺ calcdfor C ₂₆ H ₂₀ N ₄ O:405.1707；found:405.1710.

(X1-4,C ₂₆ H ₂₀ N ₄ O).White sold；Yield 72％；m.p.344.2-356℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.94(s,1H),11.80(s,1H),8.97(s,1H),8.64(s,1H),8.48(d,J＝7.9 Hz,1H),8.25–8.20(m,2H),7.74–7.65(m,3H),7.61(d,J＝7.5 Hz,3H),7.54(d,J＝7.6 Hz,1H),7.35(dt,J＝12.9,7.5 Hz,2H),7.26(d,J＝7.5 Hz,1H),2.38(s,3H)； ¹³ C NMR(126 MHz,DMSO)δ161.24,148.38,141.61,140.97,139.03,138.08,137.38,134.53,134.51,130.73,129.92,129.05,128.98,128.84,128.77,127.24,124.69,122.22,121.19,120.37,114.08,112.74,20.94；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₂₀ N ₄ O:405.1708；found:405.1710.

(X1-5,C ₂₆ H ₂₀ N ₄ O ₂ ).White sold；Yield 70％；m.p.354.1-366.9℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.93(s,1H),11.68(s,1H),8.95(s,1H),8.60(s,1H),8.48(d,J＝7.9 Hz,1H),8.22(dt,J＝6.4,1.3 Hz,2H),7.75–7.65(m,5H),7.65–7.57(m,2H),7.37–7.31(m,1H),7.07–7.02(m,2H),3.82(s,3H)； ¹³ C NMR(126 MHz,DMSO)δ161.03,160.80,148.21,141.60,140.93,139.15,137.40,134.48,129.93,129.04,128.97,128.85,128.73,127.09,122.20,121.19,120.35,114.37,113.94,112.74,55.32；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd forC ₂₆ H ₂₀ N ₄ O ₂ :421.1655；found:421.1659.

(X1-6,C ₂₆ H ₂₀ N ₄ O ₂ ).White sold；Yield 66％；m.p.352.4-367.7℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.92(d,J＝3.6 Hz,2H),8.96(d,J＝13.9 Hz,2H),8.47(d,J＝7.8 Hz,1H),8.24(dt,J＝6.3,1.3 Hz,2H),7.95(dd,J＝7.7,1.8 Hz,1H),7.69(q,J＝8.2,7.8 Hz,3H),7.65–7.57(m,2H),7.43(ddd,J＝8.7,7.2,1.8 Hz,1H),7.34(t,J＝7.3 Hz,1H),7.13(d,J＝8.3 Hz,1H),7.05(t,J＝7.5 Hz,1H),3.89(s,3H)； ¹³ C NMR(126 MHz,DMSO)δ161.42,157.80,143.58,141.59,140.96,139.25,137.41,134.49,131.44,129.03,129.00,128.81,128.73,125.72,122.65,121.19,120.74,120.33,114.07,112.74,111.85,55.64；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₂₀ N ₄ O ₂ :421.1656；found:421.1659.

(X1-7,C ₂₆ H ₂₀ N ₄ O ₂ ).Yellow sold；Yield 73％；m.p.358.5-367.6℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.95(s,1H),11.82(s,1H),8.96(s,1H),8.65(s,1H),8.48(d,J＝7.9 Hz,1H),8.23(dd,J＝7.2,1.7 Hz,2H),7.74–7.57(m,5H),7.40(t,J＝7.9 Hz,1H),7.36–7.31(m,3H),7.06–7.00(m,1H),3.83(s,3H)； ¹³ C NMR(126 MHz,DMSO)δ161.24,159.57,148.27,141.61,140.98,138.97,137.36,135.97,134.54,129.99,129.92,129.05,129.00,128.98,128.84,128.77,122.22,121.18,120.37,120.04,116.15,114.08,112.75,111.28,55.19；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₂₀ N ₄ O ₂ :421.1657；found:421.1659.

(X1-8,C ₂₇ H ₂₂ N ₄ O ₃ ).Yellow sold；Yield 71％；m.p.350.6-363.7℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.93(s,1H),11.68(s,1H),8.94(s,1H),8.58(s,1H),8.48(d,J＝7.9 Hz,1H),8.25–8.20(m,2H),7.73–7.65(m,3H),7.65–7.56(m,2H),7.40(d,J＝1.9 Hz,1H),7.34(t,J＝7.5 Hz,1H),7.24(dd,J＝8.3,1.9 Hz,1H),7.05(d,J＝8.3 Hz,1H),3.83(d,J＝15.1 Hz,6H)； ¹³ C NMR(126 MHz,DMSO)δ161.01,150.71,149.09,148.62,141.62,140.93,139.12,137.40,134.49,129.94,129.06,128.97,128.86,128.77,127.21,122.22,121.90,121.19,120.37,113.92,112.75,111.53,108.32,55.47；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₇ H ₂₂ N ₄ O ₃ :451.1762；found:451.1765.

(X1-9,C ₂₈ H ₂₄ N ₄ O ₄ ).Yellow sold；Yield 64％；m.p.345.8-351.5℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.11(s,1H),10.92(s,1H),8.10(s,1H),7.75(s,1H),7.64(d,J＝7.9 Hz,1H),7.41–7.36(m,2H),6.89–6.72(m,5H),6.50(t,J＝7.5 Hz,1H),6.23(s,2H),2.87(s,3H),1.65(p,J＝1.8 Hz,7H)； ¹³ C NMR(126 MHz,DMSO)δ161.08,153.24,148.50,141.62,140.93,139.16,138.98,137.37,134.52,129.99,129.95,129.08,128.96,128.86,128.79,122.24,121.18,120.39,113.99,112.76,104.31,60.16,55.95；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd forC ₂₈ H ₂₄ N ₄ O ₄ :481.1866；found:481.1870.

(X1-10,C ₂₅ H ₁₅ F ₃ N ₄ O).Yellow sold；Yield 68％；m.p.347.4-365℃； ¹ H NMR(500MHz,DMSO-d6)δ12.02(s,1H),11.96(s,1H),8.97(s,1H),8.63(s,1H),8.48(d,J＝7.9 Hz,1H),8.21(d,J＝7.5 Hz,2H),7.74–7.68(m,3H),7.61(q,J＝7.6 Hz,2H),7.34(t,J＝7.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.48,145.00,141.60,141.02,138.72,137.34,134.60,129.89,129.08,128.98,128.83,122.22,121.16,120.41,114.27,112.75,111.31,111.13；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₅ F ₃ N ₄ O:445.1267；found:445.1271.

(X1-11,C ₂₅ H ₁₆ F ₂ N ₄ O).Yellow sold；Yield 73％；m.p.348.5-365.2℃； ¹ H NMR(500MHz,DMSO-d6)δ12.05(s,1H),11.94(s,1H),8.96(s,1H),8.90(d,J＝2.3 Hz,1H),8.48(d,J＝7.8 Hz,1H),8.25–8.20(m,2H),8.06(td,J＝8.6,6.6 Hz,1H),7.74–7.65(m,4H),7.65–7.57(m,2H),7.43–7.31(m,2H),7.23(td,J＝8.5,2.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ164.25,162.26,162.17,161.54,159.94,141.60,141.04,140.27,140.24,138.91,137.35,134.59,129.89,129.04,129.02,128.84,128.77,128.06,128.01,127.97,122.20,121.18,120.38,119.09,119.03,114.24,112.76,112.71,112.54,104.70,104.50,104.29；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₆ F ₂ N ₄ O:427.1364；found:427.1365.

(X1-12,C ₂₅ H ₁₆ F ₂ N ₄ O).Yellow sold；Yield 71％；m.p.346.1-364.4℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(d,J＝15.2 Hz,2H),8.97(s,1H),8.66(s,1H),8.48(d,J＝7.9 Hz,1H),8.24–8.19(m,2H),7.79(ddd,J＝11.5,7.9,2.0 Hz,1H),7.74–7.51(m,8H),7.37–7.31(m,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.37,146.07,141.61,141.00,138.84,137.36,134.57,132.46,132.42,129.90,129.06,128.98,128.84,128.78,124.39,124.36,122.21,121.17,120.39,118.21,118.07,115.28,114.18,112.75；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd forC ₂₅ H ₁₆ F ₂ N ₄ O:427.1362；found:427.1365.

(X1-13,C ₂₅ H ₁₇ FN ₄ O).Yellow sold；Yield 65％；m.p.352.5-366.7℃； ¹ H NMR(500MHz,DMSO-d6)δ12.06(s,1H),11.95(s,1H),8.96(d,J＝11.1 Hz,2H),8.48(d,J＝7.9 Hz,1H),8.26–8.21(m,2H),8.03(td,J＝7.7,1.9 Hz,1H),7.74–7.65(m,3H),7.65–7.57(m,2H),7.50(tdd,J＝7.5,5.3,1.8 Hz,1H),7.33(dt,J＝13.9,7.6 Hz,3H)； ¹³ C NMR(126 MHz,DMSO)δ161.86,161.55,159.88,141.61,141.03,140.99,138.95,137.35,134.58,131.90,129.89,129.03,128.84,128.77,126.50,126.47,124.95,124.93,122.21,122.15,121.18,120.38,116.12,115.96,114.25,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ FN ₄ O:409.1458；found:409.1459.

(X1-14,C ₂₅ H ₁₇ FN ₄ O).Yellow sold；Yield 69％；m.p.357.8-367.7℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(d,J＝18.0 Hz,2H),8.97(s,1H),8.69(d,J＝2.6 Hz,1H),8.48(d,J＝7.9 Hz,1H),8.24–8.19(m,2H),7.74–7.65(m,3H),7.64–7.50(m,6H),7.38–7.26(m,2H)； ¹³ C NMR(126 MHz,DMSO)δ163.43,161.40,146.97,146.95,141.61,141.02,138.87,137.19,137.13,134.59,131.02,130.95,129.91,129.07,129.00,128.99,128.85,128.79,123.55,123.52,122.23,121.18,120.40,116.84,116.67,114.19,113.03,112.85,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ FN ₄ O:409.1456；found:409.1459.

(X1-15,C ₂₅ H ₁₇ FN ₄ O).White sold；Yield 73％；m.p.344.8-363.5℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(s,1H),11.82(s,1H),8.96(s,1H),8.68(s,1H),8.22(d,J＝7.5 Hz,2H),7.85–7.81(m,2H),7.69(q,J＝8.3,7.6 Hz,3H),7.62(t,J＝7.8 Hz,2H),7.33(q,J＝8.7,7.8 Hz,3H)； ¹³ C NMR(126 MHz,DMSO)δ164.07,162.10,161.25,147.19,141.61,140.98,138.98,137.37,134.54,131.19,131.16,129.91,129.29,129.22,129.05,128.97,128.84,128.77,122.21,121.18,120.37,116.04,115.86,114.08,112.75；HRMS(ESI-MS)m/z:[M+H] ⁺ calcdfor C ₂₅ H ₁₇ FN ₄ O:409.1456；found:409.1459.

(X1-16,C ₂₅ H ₁₇ CN ₄ O).Yellow sold；Yield 71％；m.p.345-365℃； ¹ H NMR(500 MHz,DMSO-d6)δ12.19(s,1H),11.94(s,1H),9.08(d,J＝2.6 Hz,1H),8.97(s,1H),8.48(d,J＝7.9Hz,1H),8.25–8.20(m,3H),8.10(dd,J＝5.9,3.6 Hz,1H),7.70(q,J＝8.1,7.6 Hz,4H),7.65–7.51(m,4H),7.47(dp,J＝7.2,3.7 Hz,2H),7.34(t,J＝7.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.76,144.08,141.60,141.09,139.02,137.37,134.61,133.22,132.03,131.35,129.94,129.85,129.04,128.84,128.77,127.59,127.06,122.21,121.18,120.38,114.34,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ CN ₄ O:425.1160；found:425.1164.

(X1-17,C ₂₅ H ₁₇ CN ₄ O).Yellow sold；Yield 76％；m.p.351.6-365.6℃； ¹ H NMR(500MHz,DMSO-d6)δ11.95(s,1H),11.88(s,1H),8.97(s,1H),8.67(s,1H),8.48(d,J＝8.0 Hz,1H),8.22(d,J＝7.7 Hz,2H),7.79(d,J＝8.0 Hz,2H),7.73–7.48(m,7H),7.34(d,J＝7.7 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.32,147.00,141.61,141.01,138.91,137.36,134.57,134.42,133.52,129.91,129.07,128.99,128.93,128.85,128.79,128.74,122.22,121.18,120.39,114.15,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ CN ₄ O:425.1161；found:425.1164.

(X1-18,C ₂₅ H ₁₇ CN ₄ O).Yellow sold；Yield 75％；m.p.356.2-367.2℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(d,J＝12.1 Hz,2H),8.97(s,1H),8.67(s,1H),8.48(d,J＝7.8 Hz,1H),8.22(dt,J＝6.3,1.3 Hz,2H),7.82(d,J＝2.0 Hz,1H),7.74–7.65(m,4H),7.65–7.57(m,2H),7.55–7.49(m,2H),7.34(t,J＝7.3 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.41,146.65,141.61,141.03,138.84,137.35,136.82,134.59,133.68,130.82,129.91,129.63,129.07,128.98,128.84,128.79,126.17,125.92,122.23,121.18,120.40,114.22,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ CN ₄ O:425.1161；found:425.1164.

(X1-19,C ₂₅ H ₁₇ B _r N ₄ O).White sold；Yield 67％；m.p.351.6-365.8℃； ¹ H NMR(500MHz,DMSO-d6)δ11.95(s,1H),11.88(s,1H),8.97(s,1H),8.66(d,J＝2.2 Hz,1H),8.48(d,J＝7.9 Hz,1H),8.24–8.19(m,2H),7.75–7.65(m,7H),7.64–7.58(m,2H),7.34(t,J＝7.5Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.32,147.08,141.61,141.00,138.91,137.36,134.57,133.86,131.89,129.91,129.06,128.98,128.96,128.85,128.78,123.20,122.22,121.18,120.39,114.16,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ B _r N ₄ O:469.0656；found:469.0659.

(X1-20,C ₂₅ H ₁₇ B _r N ₄ O).Yellow sold；Yield 69％；m.p.350.2-365℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(d,J＝13.5 Hz,2H),8.97(s,1H),8.65(s,1H),8.49(d,J＝7.8 Hz,1H),8.24–8.19(m,2H),7.96(t,J＝1.8 Hz,1H),7.78–7.57(m,8H),7.45(t,J＝7.8 Hz,1H),7.38–7.31(m,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.40,146.53,141.60,141.02,138.83,137.35,137.03,134.58,132.49,131.07,129.90,129.06,128.98,128.83,128.78,126.35,122.23,122.21,121.18,120.39,114.22,112.75；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd forC ₂₅ H ₁₇ B _r N ₄ O:469.0656；found:469.0659.

(X1-21,C ₂₅ H ₁₇ B _r N ₄ O).Yellow sold；Yield 67％；m.p.338.2-355.3℃； ¹ H NMR(500MHz,DMSO-d6)δ12.23(s,1H),11.94(s,1H),9.02(s,1H),8.97(s,1H),8.48(d,J＝7.9 Hz,1H),8.22(d,J＝7.5 Hz,2H),8.08(dd,J＝7.9,1.7 Hz,1H),7.70(dq,J＝12.5,7.5,5.8 Hz,4H),7.61(dt,J＝9.5,7.5 Hz,2H),7.50(t,J＝7.5 Hz,1H),7.41–7.31(m,2H)； ¹³ C NMR(126 MHz,DMSO)δ161.84,146.35,141.60,141.11,139.07,137.39,134.61,133.57,133.19,131.61,129.84,129.06,129.04,128.85,128.78,128.08,127.49,123.53,122.21,121.19,120.39,114.36,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ B _r N ₄ O:469.0655；found:469.0659.

(X1-22,C ₂₅ H ₁₈ N ₄ O ₂ ).Yellow sold；Yield 69％；m.p.353.8-366.1℃； ¹ H NMR(500MHz,DMSO-d6)δ11.27(s,1H),11.12(s,1H),10.70(s,1H),8.13(s,1H),8.06(s,1H),7.63(d,J＝7.9 Hz,1H),7.42–7.36(m,2H),6.90–6.82(m,3H),6.81–6.73(m,2H),6.69(dd,J＝7.7,1.7 Hz,1H),6.54–6.45(m,2H),6.15–6.07(m,2H)； ¹³ C NMR(126 MHz,DMSO)δ161.32,157.61,149.26,141.60,141.13,138.63,137.35,134.61,131.28,129.86,129.81,129.07,129.02,128.83,128.80,122.19,121.17,120.41,119.38,118.80,116.48,114.26,112.78；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₈ N ₄ O ₂ :407.1501；found:407.1503.

(X1-23,C ₂₅ H ₁₈ N ₄ O ₂ ).Yellow sold；Yield 79％；m.p.334.1-347.4℃； ¹ H NMR(500MHz,DMSO-d6)δ11.94(s,1H),11.77(s,1H),9.65(s,1H),8.96(s,1H),8.59(s,1H),8.48(d,J＝7.9 Hz,1H),8.24–8.19(m,2H),7.74–7.65(m,3H),7.65–7.57(m,2H),7.34(t,J＝7.5Hz,1H),7.31–7.24(m,2H),7.15(d,J＝7.5 Hz,1H),6.84(dd,J＝8.4,2.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.23,157.72,148.39,141.61,140.99,139.04,137.38,135.84,134.54,129.92,129.05,128.99,128.83,122.21,121.19,120.37,118.81,117.37,114.07,112.75,112.72；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₈ N ₄ O ₂ :407.1499；found:407.1503.

(X1-24,C ₂₅ H ₁₈ N ₄ O ₂ ).Yellow sold；Yield 70％；m.p.341.7-356.4℃； ¹ H NMR(500MHz,DMSO-d6)δ11.92(s,1H),11.62(s,1H),9.93(s,1H),8.94(s,1H),8.55(s,1H),8.47(d,J＝7.9 Hz,1H),8.24–8.19(m,2H),7.73–7.68(m,1H),7.62–7.56(m,4H),7.34(ddd,J＝7.9,7.0,1.0 Hz,1H),6.89–6.84(m,2H)； ¹³ C NMR(126 MHz,DMSO)δ160.94,159.39,148.60,141.60,140.91,139.22,137.41,134.46,129.92,129.03,128.97,128.90,128.74,125.52,122.20,121.19,120.33,115.74,113.89,112.73；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd forC ₂₅ H ₁₈ N ₄ O ₂ :407.1498；found:407.1503.

(X1-25,C ₂₅ H ₁₈ N ₄ O ₃ ).Yellow sold；Yield 76％；m.p.332.7-345.3℃； ¹ H NMR(500MHz,DMSO-d6)δ11.93(d,J＝14.0 Hz,2H),11.72(s,1H),9.98(s,1H),8.94(s,1H),8.75(s,1H),8.47(d,J＝7.9 Hz,1H),8.25–8.20(m,2H),7.74–7.65(m,3H),7.61(q,J＝7.6 Hz,2H),7.37–7.26(m,2H),6.38(dd,J＝8.3,2.3 Hz,1H),6.34(d,J＝2.3 Hz,1H)； ¹³ C NMR(126MHz,DMSO)δ160.96,160.66,159.63,150.07,141.59,141.07,138.83,137.38,134.53,131.54,129.86,129.05,129.01,128.83,128.77,122.18,121.17,120.37,114.05,112.76,110.71,107.71,102.73；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₈ N ₄ O ₃ :423.1445；found:423.1452.

(X1-26,C ₂₅ H ₁₈ N ₄ O ₃ ).Yellow sold；Yield 72％；m.p.331.4-331.8℃； ¹ H NMR(500MHz,DMSO-d6)δ11.92(s,1H),11.59(s,1H),9.39(s,1H),9.31(s,1H),8.94(s,1H),8.47(d,J＝7.2 Hz,2H),8.24–8.19(m,2H),7.73–7.65(m,3H),7.65–7.56(m,2H),7.31(dd,J＝8.0,1.5 Hz,1H),6.98(dd,J＝8.2,2.0 Hz,1H),6.81(d,J＝8.0 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ160.92,148.80,147.96,145.76,141.60,140.92,139.26,137.41,134.45,129.93,129.03,128.97,128.86,128.75,125.99,122.20,121.20,120.56,120.34,115.63,113.87,112.82,112.74；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₈ N ₄ O ₃ :423.1446；found:423.1452.

(X1-27,C ₂₅ H ₁₈ N ₄ O ₃ ).Greyish-green sold；Yield 75％；m.p.331.4-331.5℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.99(s,3H),8.97(s,1H),8.86(s,1H),8.47(d,J＝7.9 Hz,1H),8.23(d,J＝7.5 Hz,2H),7.74–7.66(m,3H),7.61(q,J＝7.7 Hz,2H),7.35(t,J＝7.5 Hz,1H),6.95(d,J＝7.7 Hz,1H),6.87(d,J＝7.8 Hz,1H),6.76(t,J＝7.8 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.35,149.96,146.29,145.73,141.63,141.17,138.62,137.36,134.64,129.88,129.10,129.04,128.86,128.82,122.20,121.19,120.43,120.31,119.17,118.88,117.36,114.30,112.81；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₈ N ₄ O ₃ :423.1448；found:423.1452.

(X1-28,C ₂₅ H ₁₈ N ₄ O ₄ ).Greyish-green sold；Yield 66％；m.p.332.4-337.5℃； ¹ H NMR(500 MHz,DMSO-d6)δ11.95(s,3H),8.95(s,1H),8.73(s,1H),8.47(d,J＝7.9 Hz,1H),8.22(d,J＝7.5 Hz,2H),7.74–7.65(m,4H),7.61(q,J＝7.7 Hz,3H),7.34(t,J＝7.5 Hz,1H),6.77(d,J＝8.4 Hz,1H),6.41(d,J＝8.4 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.01,150.97,148.77,147.60,141.61,141.11,138.80,137.38,134.55,132.82,129.87,129.08,129.02,128.96,128.85,122.19,121.19,120.39,114.10,112.78,111.02,107.71；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₈ N ₄ O ₄ :439.1398；found:439.1401.

(X1-29,C ₂₅ H ₁₇ N ₅ O ₃ ).Yellow sold；Yield 67％；m.p.358.1-369℃； ¹ H NMR(500 MHz,DMSO-d6)δ12.03(s,1H),11.96(s,1H),8.99(s,1H),8.81(s,1H),8.58(t,J＝2.0 Hz,1H),8.49(d,J＝7.9 Hz,1H),8.28(dd,J＝8.2,2.3 Hz,1H),8.25–8.20(m,2H),8.17(d,J＝7.7 Hz,1H),7.78(t,J＝7.9 Hz,1H),7.70(q,J＝8.1,7.5 Hz,3H),7.62(td,J＝7.2,5.5 Hz,2H),7.35(t,J＝7.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.49,148.23,145.93,141.60,141.04,138.70,137.34,136.39,134.62,133.41,130.52,129.90,129.08,128.98,128.84,128.79,124.15,122.23,121.17,120.80,120.40,114.29,112.75；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd forC ₂₅ H ₁₇ N ₅ O ₃ :436.1401；found:436.1404.

(X1-30,C ₂₅ H ₁₇ N ₅ O ₃ ).Yellow sold；Yield 69％；m.p.343.8-345.8℃； ¹ H NMR(500MHz,DMSO-d6)δ12.24(s,1H),11.95(s,1H),9.07(s,1H),8.98(s,1H),8.48(d,J＝7.9 Hz,1H),8.20(t,J＝7.2 Hz,4H),8.07(d,J＝8.1 Hz,1H),7.84(t,J＝7.6 Hz,1H),7.70(dt,J＝10.9,8.0 Hz,5H),7.61(dt,J＝10.2,7.4 Hz,3H),7.34(t,J＝7.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.89,148.40,143.05,141.61,141.12,138.87,137.35,134.65,133.66,130.60,129.85,129.06,129.02,128.89,128.85,128.79,128.06,124.59,122.22,121.18,120.40,114.44,112.77；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ N ₅ O ₃ :436.1403；found:436.1404.

(X1-31,C ₂₅ H ₁₇ N ₅ O ₃ ).Yellow sold；Yield 72％；m.p.331-345.8℃； ¹ H NMR(500 MHz,DMSO-d6)δ12.10(s,1H),11.97(s,1H),8.99(s,1H),8.80(s,1H),8.49(d,J＝7.9 Hz,1H),8.33(d,J＝8.3 Hz,2H),8.22(d,J＝7.5 Hz,2H),8.02(d,J＝8.4 Hz,2H),7.70(q,J＝7.9,7.5Hz,3H),7.62(t,J＝7.1 Hz,2H),7.35(t,J＝7.6 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.59,147.78,145.86,141.62,141.09,140.92,138.67,137.33,134.66,129.91,129.11,129.01,128.87,128.83,127.98,124.15,122.25,121.18,120.44,114.39,112.78；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₅ H ₁₇ N ₅ O ₃ :436.1403；found:436.1404.

(X1-32,C ₂₆ H ₁₇ F ₃ N ₄ O).Yellow sold；Yield 67％；m.p.357.6-366.9℃； ¹ H NMR(500MHz,DMSO-d6)δ12.34(s,1H),11.93(s,1H),9.07(d,J＝2.4 Hz,1H),8.98(s,1H),8.48(d,J＝7.9 Hz,1H),8.30(d,J＝7.8 Hz,1H),8.22–8.17(m,2H),7.84–7.76(m,2H),7.74–7.57(m,6H),7.38–7.31(m,1H)； ¹³ C NMR(126 MHz,DMSO)δ162.11,143.07,141.60,141.18,139.07,137.40,134.66,132.82,132.72,129.80,129.04,128.84,127.16,125.88,122.21,121.18,120.39,114.49,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₁₇ F ₃ N ₄ O:459.1425；found:459.1427.

(X1-33,C ₂₆ H ₁₇ F ₃ N ₄ O).Yellow sold；Yield 72％；m.p.347-364.3℃； ¹ H NMR(500MHz,DMSO-d6)δ11.97(d,J＝6.6 Hz,2H),8.98(s,1H),8.77(s,1H),8.49(d,J＝7.8 Hz,1H),8.25–8.20(m,2H),8.10(s,1H),8.05(d,J＝7.7 Hz,1H),7.81(d,J＝7.8 Hz,1H),7.77–7.65(m,4H),7.62(t,J＝7.6 Hz,2H),7.34(t,J＝7.5 Hz,1H)； ¹³ C NMR(126 MHz,DMSO)δ161.46,146.63,141.62,141.04,138.78,137.35,135.73,134.61,131.15,130.14,129.80,129.54,129.00,128.85,128.80,126.24,123.00,122.91,122.88,122.24,121.18,120.40,114.24,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₁₇ F ₃ N ₄ O:459.1424；found:459.1427.

(X1-34,C ₂₆ H ₁₇ F ₃ N ₄ O).Yellow sold；Yield 69％；m.p.329.8-338.5℃； ¹ H NMR(500

MHz,DMSO-d6)δ12.00(s,1H),11.96(s,1H),8.98(s,1H),8.77(d,J＝2.3Hz,1H),8.49(d,J＝7.9Hz,1H),8.25–8.20(m,2H),7.99(d,J＝8.0Hz,2H),7.84(d,J＝8.3Hz,2H),7.74–

7.65(m,3H),7.65–7.57(m,2H),7.35(t,J＝7.5Hz,1H)； ¹³ C NMR(126MHz,DMSO)δ

161.49,146.58,141.62,141.05,138.79,138.56,137.35,134.62,129.97,129.91,129.07,129.03,

128.99,128.85,128.79,127.66,125.79,125.75,125.72,122.22,121.18,120.40,114.27,112.76；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₁₇ F ₃ N ₄ O:459.1425；found:459.1427.

(X1-35,C ₂₆ H ₁₇ N ₅ O).Yellow sold；Yield 72％；m.p.342.8-350.2℃； ¹ H NMR(500

MHz,DMSO-d6)δ12.04(s,1H),11.96(s,1H),8.98(s,1H),8.74(s,1H),8.48(d,J＝7.9Hz,1H),8.24–8.19(m,2H),7.94(s,4H),7.74–7.57(m,5H),7.34(t,J＝7.5Hz,1H)； ¹³ C NMR

(126MHz,DMSO)δ161.51,146.32,141.61,141.05,139.06,138.72,137.33,134.63,132.79,129.89,129.07,128.99,128.84,128.80,127.61,122.22,121.17,120.41,118.73,114.32,112.76,

111.77；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₁₇ N ₅ O:416.1503；found:416.1506.

(X1-36,C ₂₆ H ₁₇ N ₅ O).White sold；Yield 69％；m.p.353.6-366.2℃； ¹ H NMR(500MHz,DMSO-d6)δ12.02(s,1H),11.96(s,1H),8.98(s,1H),8.71(s,1H),8.48(d,J＝7.8Hz,1H),

8.25–8.20(m,2H),8.11(d,J＝8.0Hz,1H),7.90(d,J＝7.6Hz,1H),7.74–7.65(m,4H),

7.61(q,J＝7.2Hz,2H),7.34(t,J＝7.5Hz,1H)； ¹³ C NMR(126MHz,DMSO)δ161.49,146.02,141.60,141.03,138.79,137.35,135.92,134.60,133.17,131.17,130.50,130.17,129.89,

129.07,129.02,128.99,128.84,128.79,122.23,121.17,120.40,118.45,114.27,112.76,112.05；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₁₇ N ₅ O:416.1504；found:416.1506.

(X1-37,C ₂₆ H ₁₇ N ₅ O).Greyish-green sold；Yield 73％；m.p.360.2-367.7℃； ¹ H NMR(500MHz,DMSO-d6)δ12.38(s,1H),11.96(s,1H),9.07(s,1H),8.98(s,1H),8.48(d,J＝7.9Hz,1H),8.21(ddd,J＝8.1,3.8,1.3Hz,3H),7.93(dd,J＝7.8,1.3Hz,1H),7.82(td,J＝7.8,1.4Hz,1H),7.74–7.66(m,3H),7.66–7.57(m,3H),7.38–7.31(m,1H)； ¹³ C NMR(126MHz,DMSO)δ161.96,143.31,141.60,141.15,138.92,137.50,134.66,133.53,133.49,130.25,129.82,129.05,128.84,128.78,125.82,122.21,121.18,120.39,117.23,114.47,112.77,110.79；HRMS(ESI-MS)m/z:[M+H] ⁺ calcd for C ₂₆ H ₁₇ N ₅ O:416.1503；found:416.1506.

example 38: alpha-glucosidase inhibition activity test of beta-carboline derivative

1. Preparation of reagents and Standard solutions

(1) 100mM phosphate buffer (PBS, pH 6.8): weighing a certain mass of potassium dihydrogen phosphate and disodium hydrogen phosphate, dissolving with ultrapure water, and dissolving a diluting reagent.

(2) Preparing an alpha-glucosidase solution: adding a proper amount of 100mM PBS into enzyme with the enzyme activity of 100U to prepare the enzyme with the working concentration of 0.05U/mL, and sub-packaging and freezing.

(3) Preparing a substrate: precisely weighing a proper amount of 4-nitrophenyl-D-glucopyranoside (PNPG), adding 100mM PBS solution for dissolution, preparing a substrate buffer solution with the content of about 0.25mM, and uniformly swirling for fresh use in animal experiments.

The preparation of test medicine, namely, precisely weighing a proper amount of the medicine to be tested, dissolving and preparing into 10mM stock solution by using DMSO, and storing at the shade-avoiding temperature of-20 ℃. In the test, the DMSO solutions are diluted to different levels (0-200 mu M) and the DMSO content is less than 5%.

2. Experimental procedure

(1) 10. Mu.L of alpha-glucosidase at a concentration of 0.05U/mL, 130. Mu.L (pH 6.8) of phosphate buffer at a concentration of one hundred mM, 10. Mu.L of a compound (the beta-carboline derivative X1-1 to X1-37 prepared in example 1) at different concentrations, a blank group, in which 10. Mu.L of a DMSO solution at a concentration of more than 5% was used as a positive control, four multiplex wells were placed in parallel in each group, and the enzyme reaction system was incubated in an microplate reader at 37℃for 10 minutes.

(2) Then, about 50. Mu.L of the substrate PNPG was added to the enzyme reaction system to activate the enzyme reaction system, and the microplate was placed in a microplate reader at 37℃for further incubation for 15min, three hours were used on average during the incubation, each timeThe average length was measured once at 405nm, and the reading was recorded as OD ₁ 、OD ₂ 、OD ₃ 。

(3) The activity of alpha-glucosidase in a test substance can be determined as follows:

inhibition (%) = [ (OD) ₃ -OD)-(OD ₁ -OD)]/OD ₃ -OD×100％

Where OD is the highest absorbance value in the blank time control, data processing: experimental data were processed by MS Excel analysis and then the compound inhibition content value (IC) at 50% inhibition of α -glucosidase was obtained by origin9.1 operation ₅₀ )，IC ₅₀ The content of the test substance required for limiting the activity of the alpha-glucosidase to 50% under the above-specified test conditions is shown.

3. Analysis of results

The prepared substance is subjected to the effectiveness evaluation of alpha-glucosidase by adopting an in-vitro enzymology test, and the detection result is shown in table 2:

TABLE 2 in vitro inhibition function study of Compounds obtained after screening with alpha-glucosidase

^a The inhibition rate of the test compound is lower than 50% at 100 mu M;

^b the values are mean±s for the results of three independent experiments.

Wherein the positive control drug Acarbose (Acarbose) IC ₅₀ 640. Mu.M. As shown in fig. 1a, 1b and 1c, fig. 1a is a graph of half-maximal inhibitory concentration of X1-25; FIG. 1b is a graph of half inhibition concentration of X1-26; FIG. 1c is a graph of half inhibition concentration of X1-27; the beta-carboline derivatives are X1-25, X1-26 and X1-27, and all show the strongest alpha-glucosidase inhibition effect, IC ₅₀ Value scoreThe concentration of the alpha-glucosidase inhibitor is 2.67+/-0.54 mu M, 0.65+/-0.01 mu M and 1.57 mu M, which are 239-985 times of acarbose, so that the alpha-glucosidase inhibitor can be used for treating or preventing diabetes.

The results show that these small molecule compounds exhibit a relatively strong binding affinity when interacting with alpha-glucosidase. It can be deduced that the beta-carboline skeleton structure plays a very important role in better alpha-glucosidase inhibitory activity of the compound, and the compound such as X1-25, X1-26, X1-27 and the like can be used as an alpha-glucosidase inhibitor for treating or preventing diabetes.

Example 39 enzyme kinetic assay

The inhibition activity kinetics evaluation of the alpha-glucosidase is carried out on the synthesized active compound by adopting an in-vitro enzyme kinetics experiment:

1. preparation of reagents and Standard solutions

(2) Preparing an alpha-glucosidase solution: the enzyme with the enzyme activity of 100U is added with a proper amount of 100mM PBS to prepare the enzyme with the working concentration of 0.0375U/mL, 0.05U/mL, 0.0625U/mL and 0.075U/mL respectively, and the enzyme is split-packed and frozen.

(3) Preparing a substrate: accurately weighing a proper amount of 4-nitrophenyl-D-glucopyranoside (PNPG), adding into 100mM PBS solution, preparing a substrate buffer solution with the content of about 0.25mM, and uniformly swirling for fresh use in animal experiments.

The preparation method of the test medicine comprises accurately weighing the medicine to be tested as the optimal amount, dissolving with DMSO solution, preparing into about 10mM stock solution, and storing at-20deg.C in dark place. Before the experiment, the solution was diluted to different desired concentrations (0-200. Mu.M) with DMSO content equal to 5%.

2. Experimental procedure

(1) To 96-well plates, 10. Mu.L of alpha-glucosidase 0.0375U/mL, 0.05U/mL, 0.0625U/mL, 0.075U/mL, 130. Mu.L (pH 6.8) of phosphate buffer containing one hundred mM, 10. Mu.L of the derivative (beta-carboline compound X1-26 prepared in example 26) were added, respectively, 10. Mu.L of DMSO was used as a blank to replace 10. Mu.L of the derivative, acarbose was used as a positive control, four wells were designed in parallel in each group, and the enzyme reaction system was incubated in an ELISA at 37℃for 10min.

(2) Then, 50. Mu.L of about 0.25mM of the substrate PNPG was added to the enzyme reaction system to perform the enzyme reaction, and the microplate was placed in a microplate reader at 37℃for further incubation for about 15 minutes, and an average of 3 hours was allocated during the incubation, and the average length of each period was once read at 405nm, and the reading was recorded as OD ₁ 、OD ₂ 、OD ₃ 。

(3) And (3) data processing: the data were analyzed using the Origins data processing software, and the reaction rate of the enzyme reaction system was ΔOD/min.

3. Analysis of results

The results of the enzyme kinetic inhibition type evaluation experiment are shown in FIG. 2. As can be seen from FIG. 2, the inhibition of the enzyme activity by non-covalent binding of the inhibitor to the enzyme is a reversible inhibition.

Example 40: substrate kinetics experiments

The in vitro substrate kinetics experiment is adopted to evaluate the inhibition activity kinetics of the alpha-glucosidase of the synthesized active compound:

1. preparation of reagents and Standard solutions

(3) Preparing a substrate: accurately weighing a proper amount of 4-nitrophenyl-D-pyrrologlucoside (PNPG), adding into 100mM PBS solution, mixing into corresponding (0.25 mM, 0.5mM, 0.75mM, 1 mM) substrate buffer solution, and swirling uniformly for fresh application in animal experiments.

The formulation of the test agent is that a proper amount of the precisely weighed agent to be tested is dissolved by DMSO solution and then added into each 10mM stock solution, and the stock solution is stored at-20 ℃ in a shade-avoiding manner. The DMSO solutions were diluted to different desired levels (0-200 μm) in the experiment, with an average DMSO content of about 5%.

2. Experimental procedure

(1) 10. Mu.L of alpha-glucosidase with a content of 0.5U/mL, 130. Mu.L (pH 6.8) of phosphate buffer with a content of one hundred mM, 10. Mu.L of derivative (beta-carboline compound X1-26 prepared in example 26) with different content are respectively added into a 96-well plate, 10. Mu.L of derivative is replaced by a blank control group with 10. Mu.L of DMSO with a concentration of more than five percent, acarbose is used as a positive control, four compound wells are placed in parallel in each group, and the enzyme reaction system is put into an enzyme-labeled instrument for incubation at 37 ℃ for 10min.

(2) Then adding 50 mu L of different content of substrate PNPG into the enzyme reaction system to activate the enzyme reaction system, placing the microplate in a microplate reader at 37 ℃ and then continuously incubating for 15min, wherein the average time is about three hours during incubation, reading at 405nm for each period of time, and recording the reading as OD ₁ 、OD ₂ 、OD ₃ 。

(3) And (3) data processing: the data were analyzed using an origin data processing, and the reaction rate of the enzyme reaction system was DeltaOD/min.

3. Analysis of results

The evaluation test detection result of the substrate dynamics inhibition is shown in figure 3, and figure 3 corresponds to the substrate dynamics image of the beta-carboline derivative X1-26 for inhibiting alpha-glucosidase in vitro, and the substrate dynamics image is obtained by adopting a double reciprocal plotting method. As can be seen from fig. 3, the inhibitor drug detected is a non-competitive inhibitor and can bind not only to α -glucosidase but also to the α -glucosidase receptor.

The present invention has been described in detail with reference to the above embodiments, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.

Claims

1. A beta-carboline derivative or a pharmaceutically acceptable salt thereof, which is characterized in that the beta-carboline derivative has a structure shown in a formula (I):

2. The β -carboline derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein R is selected from substituted or unsubstituted phenyl, said substitution is mono-or polysubstituted, and said substituted group is selected from halogen, C _1～6 Alkyl, C of (2) _1～6 Alkoxy, C _1～6 Haloalkyl, hydroxy, nitro, cyano.

3. The β -carboline derivative or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R is selected from one of the following groups:

4. a method for the preparation of a β -carboline derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, comprising the steps of:

wherein, the structure of compound 1 is as follows:

5. the method according to claim 4, wherein the solvent is at least one selected from ethanol, tetrahydrofuran, pyridine, methanol and N, N-dimethylformamide.

6. The process according to claim 4, wherein the temperature of the reaction is 15 to 30 ℃.

7. The method according to claim 4, wherein the reaction further comprises a purification step after completion of the reaction.

8. A pharmaceutical composition comprising a β -carboline derivative according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof; pharmaceutically acceptable auxiliary materials.

9. An alpha-glucosidase inhibitor, comprising a beta-carboline derivative according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

10. Use of a β -carboline derivative according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 8, for the manufacture of a medicament for the treatment and/or prophylaxis of diabetes.